|Year : 2018 | Volume
| Issue : 1 | Page : 24-32
Neuroprotective effects of tenuigenin on neurobehavior, oxidative stress, and tau hyperphosphorylation induced by intracerebroventricular streptozotocin in rats
Xiao-Bo Huang, Yu-Jing Chen, Wen-Qiang Chen, Ning-Qun Wang, Xi-Ling Wu, Yan Liu
Department of Traditional Chinese Medicine, Xuanwu Hospital of Capital Medical University, 45 Changchun Street, Xicheng District, Beijing, 100053, PR China
|Date of Submission||07-Mar-2017|
|Date of Decision||16-Nov-2017|
|Date of Acceptance||25-Nov-2017|
|Date of Web Publication||18-Apr-2018|
Dr. Yu-Jing Chen
No. 45, Chang-Chun Street, Beijing 100053
Source of Support: None, Conflict of Interest: None
BACKGROUND: Tenuigenin (TEN), a major active component of the Chinese herb Polygala tenuifolia root, has been used to improve memory and cognitive function in Traditional Chinese Medicine for centuries. PURPOSE: The present study was designed to explore the possible neuroprotective effect of TEN on the streptozotocin (STZ)-induced rat model of sporadic Alzheimer's disease (sAD). METHODS: STZ was injected twice intracerebroventrically (3 mg/kg, ICV) on alternate days (day 1 and day 3) in Rats. Daily treatment with TEN (2, 4, and 8 mg/kg) starting from the first dose of STZ for 28 days. Memory-related behaviors were evaluated using the Morris water maze test. Hyperphosphorylation of tau proteins in hippocampus were measured by western blot assay. Superoxide dismutase activities, malondialdehyde, glutathione peroxidase and 4-hydroxy-2-nonenal adducts contents were also measured in the hippocampus.RESULTS: Treatment with TEN significantly improved STZ-induced cognitive damage, markedly reduced changes in malondialdehyde and 4-hydroxy-2-nonenal adducts, and significantly inhibited STZ-induced reduction in superoxide dismutase and glutathione peroxidase activities in the hippocampus. In addition, TEN decreased hyperphosphorylation of tau resulting from intracerebroventricular STZ (ICV-STZ) injection, and Nissl staining results showed that TEN has protective effects on hippocampal neurons. CONCLUSION: These results provide experimental evidence demonstrating preventive effect of TEN on cognitive dysfunction, oxidative stress, and hyperphosphorylation of tau in ICV-STZ rats. This study indicates that TEN may have beneficial effects in the treatment of neurodegenerative disorders such as AD.
Keywords: Intracerebroventricular streptozotocin, oxidative stress, tau hyperphosphorylation, tenuigenin
|How to cite this article:|
Huang XB, Chen YJ, Chen WQ, Wang NQ, Wu XL, Liu Y. Neuroprotective effects of tenuigenin on neurobehavior, oxidative stress, and tau hyperphosphorylation induced by intracerebroventricular streptozotocin in rats. Brain Circ 2018;4:24-32
|How to cite this URL:|
Huang XB, Chen YJ, Chen WQ, Wang NQ, Wu XL, Liu Y. Neuroprotective effects of tenuigenin on neurobehavior, oxidative stress, and tau hyperphosphorylation induced by intracerebroventricular streptozotocin in rats. Brain Circ [serial online] 2018 [cited 2021 Oct 25];4:24-32. Available from: http://www.braincirculation.org/text.asp?2018/4/1/24/230245
| Introduction|| |
Alzheimer's disease (AD) is an age-related neurodegenerative disease characterized by a progressive decline in cognitive functions. The exact etiology of AD has not been clearly identified, but numerous pharmacological research studies have demonstrated that oxidative stress is involved in AD-type neurodegeneration with cognitive impairment and age-related cognitive deficit. Oxidative stress is an imbalance between an oxidant (reactive oxygen species [ROS]/reactive nitrogen species [RNS]) and antioxidant defense systems. This increase in the ROS/RNS can lead to mitochondrial dysfunction, tau hyperphosphorylation, and β-amyloid (Aβ) overexpression.,,
Streptozotocin (STZ), a glucosamine- nitrosourea chemical compound, can generate a cytotoxic product when metabolized, which induces preferential damage in pancreatic β cells. Intracerebroventricular (ICV) injection of STZ in rats can produce long-term and progressive loss of memory. It also impairs cerebral glucose and energy metabolism, resulting in oxidative stress and inflammation., Furthermore, ICV-STZ leads to increased Aβ accumulation and hyperphosphorylation of tau proteins in the hippocampus., Thus, the method of ICV injection of STZ provides a useful model for studying the mechanism and therapeutic intervention of AD.
Tenuigenin (TEN), the active component of the Chinese herb Polygala tenuifolia root, has been used to improve memory and intelligence in Traditional Chinese Medicine for approximately 2000 years. [Figure 1] shows the structural formula of TEN. TEN has been reported to have various biological and pharmacological activities such as antioxidation, anti-inflammation, antidementia, and anti-aging., Pharmacological data indicate that TEN could suppress secretion of Aβ in SH-SY5Y APP 695 cells  and protect SH-SY5Y cells against the injury induced by 6-OHDA. Our previous study has shown that TEN could improve the proliferation and differentiation of hippocampal neural stem cells in vitro. It also protected hippocampal neuronal cells against damage induced by methylglyoxal through its antioxidant and antiapoptotic properties.
|Figure 1: (a) Chemical structure of tenuigenin. (b) The experimental design treatment schedule and intervals for estimation of various parameters. ICV-STZ = intracerebroventricular streptozotocin, MWM = Morris water maze, SAC = sacrificed|
Click here to view
In the present study, we usein vivo model to evaluate the protective effects of TEN against learning and memory impairments, oxidative stress, and tau protein phosphorylation induced by ICV injection of STZ.
| Materials and Methods|| |
Adult male Sprague-Dawley rats, weighing 250–300 g (obtained from Beijing Vital River Laboratory Animal Technology, Co. Ltd., China), were used in the present experiments. All rats were housed under a 12/12 h light/dark cycle and specific pathogen-free conditions. Food and water were provided ad libitum. All animal care and experimental procedures were carried out according to the National Institutes of Health Guide for Care and Use of Laboratory Animals and were approved by the Bioethics Committee of Xuan Wu Hospital of Capital Medical University.
Chemicals and reagents
TEN (purity >98%) was purchased from the National Institute of Pharmaceutical and Biological Products (Beijing, China). STZ, 4-hydroxy-2-nonenal (4-HNE), thiobarbituric acid (TBA), paraformaldehyde, hematoxylin, and eosin were purchased from Sigma (St. Louis, USA). Superoxide dismutases (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) kits were obtained from Nanjing Jiancheng Bioengineering Research Institute (Nanjing, China). Total tau, ser396-phosphorylated tau, thr181-phosphorylated tau, and β-actin-specific antibodies were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA).
Intracerebroventricular administration of streptozotocin
ICV injection of STZ was performed as previously described. Rats were anesthetized with 10% chloral hydrate (3 ml/kg, i. p.), the head was fixed in a stereotactic frame and a midline sagittal incision was made in the scalp. Two holes were drilled in the skull on both sides over the lateral ventricles using the following coordinates: 0.8 mm posterior to the bregma, 1.5 mm lateral to the sagittal suture, and 3.6 mm beneath the surface of the brain. Rats in the STZ and three TEN groups were given ICV injections of STZ (3 mg/kg bilaterally, on day 1 and 3). STZ was dissolved in citrate buffer (pH 4.4) shortly before application. Rats in the control group were given an ICV injection of the same volume of artificial cerebral spinal fluid (ACSF, 147 mM NaCl, 2.9 mM KCl, 1.6 mM MgCl2, 1.7 mM CaCl2, and 2.2 mM dextrose) on day 1 and day 3.
Animal groups and drug administration
After a week of adaption to colony room conditions, all rats were randomly divided into five groups consisting of 12 animals each (n = 12): Group I: Sham operated, vehicle-treated control (Sham); Group II: ICV-STZ-infused and vehicle-treated (ICV-STZ); Group III: ICV-STZ-infused and 2 mg/kg TEN treated (TEN 2 mg/kg); Group IV: ICV-STZ-infused and 4 mg/kg TEN treated (TEN 4 mg/kg); and Group V: ICV-STZ-infused and 8 mg/kg TEN treated (TEN 8 mg/kg). The administration was conducted once daily after the first STZ infusion until the water maze trial. Thus, the total period of administration was 28 days. The TEN dose used in this study was determined from previous studies. The animals in the Sham and STZ groups were administered with the same amount of double-distilled water. All drugs were administered by intragastric administration.
Morris water maze test
The Morris water maze (MWM) procedure is most commonly used to test the spatial memory. The water maze is a circle-shaped black pool (150 cm diameter, 40 cm high, and filled up to 30 cm with water at 28 ± 1°C). The pool is divided into four quadrants (1, 2, 3, and 4), and a platform is placed in the 4th quadrant submerged 1 cm below the water surface. Animals performed four trials daily for five consecutive days. Each trial was started by placing the animal into the tank, facing the wall of the tank. Before each trial, the releasing position was randomly selected by computer. The latency time to find the hidden platform and the swim velocity were recorded; when 120 s had elapsed, the trial was terminated automatically. The rat was allowed to stay on the platform for 5 s. If they were not able to find the platform after 120 s of swimming, they were put on the platform gently by the examiner and allowed to stay on it for 15 s. After each trial, rats were gently dried and placed in their home cage. Using the ANY-maze video tracking system (Caterpillar instrumentation Pvt.), we analyzed the path of each rat. On the 6th day, a probe trial was done by removing the platform and allowing the rats swim freely in the pool for 60 s. The results were obtained by calculating the time spent in the target quadrant where the platform had previously been located during the training phase. This parameter provided a measure of task comprehension.
After being anesthetized with 10% chloral hydrate, the rats were subjected to transaortic perfusion with phosphate-buffered saline followed by 4% paraformaldehyde to fix the brain. After fixation, the tissue was dehydrated and embedded in paraffin. Coronal sections at the level of the hippocampus were collected and cut into 4 mm thickness. Nissl staining was done as described in the protocol of the instructions. Sections were observed under light microscope (Olympus, CX-21) for histopathological changes. Six random visual fields of the hippocampal CA1 were photographed in each section. At high magnification (×400), the number of staining cells in each field was calculated, and the data were represented as the number of staining cells.
Assessment of the lipid peroxidation
Preparation of brains samples
After completing the MWM test, rats were sacrificed by decapitation and brain tissues were dissected and rinsed with ice-cold isotonic saline. The hippocampus samples were quickly separated and then homogenized with phosphate buffer solution (pH 7.4). The homogenate was centrifuged at 1500 rpm for 10 min (4°C) and aliquots of supernatant were separated and used for assays of SOD, GSH-Px, and MDA. Protein concentrations were determined in the hippocampus samples using the Bradford assay.
Activities of SOD, GSH-Px, and MDA in the hippocampus were measured according to the manufacturer's instructions.
The activity of SOD was determined by detecting its ability to inhibit the photochemical reduction of nitro blue tetrazolium (NBT). One unit (U) of SOD activity is defined as the amount of enzyme required to inhibit the rate of NBT reduction by 50%. The maximum absorbance was read at 550 nm and enzymatic activity was expressed as U/mg protein/min.
GSH-Px activity was measured by quantifying the rate of oxidation of the reduced glutathione to the oxidized glutathione, which was catalyzed by GSH-Px. One unit of GPX is defined as the amount of enzyme required to oxidize 1 nmol GSH/min. The GSH-Px activity in the supernatant was expressed as U/mg protein.
MDA is a biomarker to measure the level of lipid peroxidation. MDA can react with TBA, which produces a pink complex with a peak absorbance at 532 nm. The MDA level was expressed as nmol per mg protein.
Western blot analysis
Analyses of immunocontent of 4-HNE protein adduct, tau, and phosphorylated tau (ser396 and thr181) in the hippocampus were performed using Western blotting. Briefly, the hippocampus was homogenized in ice-cold lysis buffer containing protease and phosphatase inhibitors. The protein concentrations of the samples were determined using a BCA protein assay kit (Bio-Rad, Hercules, CA, USA). After electrophoresis, the proteins were transferred to a PVDF membrane. Then, the membranes were blocked in Tris-buffered saline mixed with Tween-20 (TBST) containing 10% skim milk for 60 min. In the incubation overnight, we used a blocking solution containing the respective antibody 4-HNE (1:2000), total tau (1:1000), primary rabbit anti-ser396-p-tau (1:1000), rabbit anti-thr181-p-tau (1:1000), and mouse anti-β-actin (1:1000). After washing with TBST, the membranes were incubated with anti-rabbit or anti-mouse secondary antibodies for 2 h at room temperature and washed again in TBST buffer. Finally, the membranes were developed using a chemiluminescence detection system and densitometry was analyzed with the ImageJ software. β-actin was used as a housekeeping protein.
Data are expressed as the mean ± standard deviation. Statistical analysis of the results was performed by applying the analysis of variance, followed by Tukey's test for biochemical parameters, and the Mann–Whitney test for behavioral observations. P < 0.01 was considered statistically significant.
| Results|| |
Effects of tenuigenin on spatial learning and memory deficits induced by intracerebroventricular injection of streptozotocin
Learning and memory functions were assessed using the MWM test, which includes the place navigation task and spatial probe test. The results showed that after 5 days of training, the escape latency to reach the submerged platform decreased significantly in experimental rats of all groups. From day 3 onward, the STZ group rats had a significant increase in the latency time required to reach the platform throughout the training period than did the Sham group rats, indicating poorer learning and memory performance (P< 0.01). Treatment with TEN significantly decreased escape latency compared with the STZ group. In addition, 8 mg/kg of TEN was more effective than 2 mg/kg and 4 mg/kg (P< 0.05, P < 0.01). The effects of 2 mg/kg and 4 mg/kg TEN on escape latency were not significantly different [Figure 2]a.
|Figure 2: (a) Effects of tenuigenin on memory deficits induced by intracerebroventricular injection of streptozotocin, as evaluated by the Morris water maze test. Streptozotocin-injected rats had longer escape latency than the Sham group, but administration of tenuigenin showed a dose-dependent reduction in escape latency as compared with the streptozotocin group. All values are expressed as mean ± standard deviation. ##P < 0.01 vs. Sham group and **P < 0.01 versus streptozotocin group. ◬P < 0.05 or ▴P < 0.01 versus tenuigenin 2 mg/kg group. (b) Comparison of the time spent in the target quadrant with other quadrants. The average time in the target quadrant was significantly reduced in the streptozotocin group as compared with the Sham group and which was reversed by tenuigenin treatments. All values are expressed as mean ± standard deviation. ##P < 0.01 versus Sham group and **P < 0.01 versus streptozotocin group. ◬P < 0.05 or ▴P < 0.01 versus tenuigenin 2 mg/kg group. ★P < 0.05 versus tenuigenin 4 mg/kg group|
Click here to view
Similar effects were observed in the spatial probe test with the platform removed. STZ treated rats had a significantly decreased amount of time spent in the target quadrant than the Sham group rats (P< 0.01), as they could not remember the precise location of the platform. However, the amount of time spent in the target quadrant by the rats in the TEN groups was much longer than that in the STZ group (P< 0.05, P < 0.01) [Figure 2]b, supporting a protective effect on learning and memory.
Effects of tenuigenin on hippocampal neurons damage induced by intracerebroventricular injection of streptozotocin
The results of Nissl staining are shown in [Figure 3]. Neuronal loss and damaged neurons were observed in the hippocampus of the STZ group. The histopathology of the injured neurons showed shrinkage with condensed nuclei and sparse Nissl bodies. In the TEN (2, 4, and 8 mg/kg) groups, it can be clearly observed that neurons had larger cell bodies, and obvious Nissl bodies were seen in most of the neurons compared with the STZ group [Figure 3]a. Our results showed that the number of Nissl staining cells decreased remarkably in the STZ group, and this effect can be attenuated by TEN in a dose-dependent manner (P< 0.05, P < 0.01) [Figure 3]b.
|Figure 3: (a) Nissl stain in the CA1 region of hippocampus. Sham group indicates the normal neurons (①). Streptozotocin group revealed neuronal loss (②). Tenuigenin markedly decreases the loss of the cells (③-⑤). Scale bar is 20 μm. (b) The number of Nissl staining cells in the hippocampal CA1 regions. Quantitative cell count revealed an increase in the number of surviving neurons in the tenuigenin groups in a dose-dependent manner. All values are expressed as mean ± standard deviation (n = 5). ##P < 0.01 versus Sham group and **P < 0.01 versus streptozotocin group. ◬P < 0.05 or ▴P < 0.01 versus tenuigenin 2 mg/kg group. ★P < 0.05 versus tenuigenin 4 mg/kg group|
Click here to view
Effects of tenuigenin on superoxide dismutases activity, glutathione peroxidase, and malondialdehyde contents in the hippocampus induced by intracerebroventricular injection of streptozotocin
To understand the antioxidant effects of TEN, the SOD activity in the hippocampus was measured and summarized in [Figure 4]a. The levels of SOD in the hippocampus decreased significantly in the STZ group compared to the Sham group (P< 0.01). TEN (2, 4, and 8 mg/kg) treatment for 28 days significantly inhibited the reduction of SOD activity in a dose-dependent manner in the hippocampus (P< 0.05, P < 0.01) [Figure 4]a.
|Figure 4: (a) Effects of tenuigenin on superoxide dismutases in the hippocampus induced by intracerebroventricular injection of streptozotocin. Streptozotocin significantly decreased the superoxide dismutases activity in the hippocampus as compared with the Sham group (##P < 0.01 vs. Sham group), whereas tenuigenin had significant effect on increasing the activity of superoxide dismutases in a dose-dependent manner (**P < 0.01 vs. streptozotocin group, ◬P < 0.05 or ▴P < 0.01 vs. tenuigenin 2 mg/kg group, ★P < 0.05 vs. tenuigenin 4mg/kg group). (b) Effects of tenuigenin on glutathione peroxidase in the hippocampus induced by intracerebroventricular injection of streptozotocin. The activity of glutathione peroxidase in the hippocampus of the streptozotocin group was significantly lowered compared with the Sham group (##P < 0.01 vs. Sham group), but treatment with tenuigenin notable restored the glutathione peroxidase activity in a dose-dependent manner (**P < 0.01 vs. streptozotocin group, ◬ P < 0.05 or ▴ P < 0.01 vs. tenuigenin 2 mg/kg group, ★P < 0.05 vs. tenuigenin 4mg/kg group). (c) Effects of tenuigenin on malondialdehyde level in the hippocampus induced by intracerebroventricular injection of streptozotocin. Intracerebroventricular - streptozotocin caused a significant increase in the levels of malondialdehyde in the hippocampus (##P < 0.01 vs. Sham group). Administration of tenuigenin significantly restored the levels of malondialdehyde. (**P < 0.01 vs. streptozotocin group, ◬P < 0.05 or ▴P < 0.01 vs. tenuigenin 2 mg/kg group)|
Click here to view
GSH-Px activity was measured to evaluate the enzymatic defense potential of the cells against the oxidative stress. As shown in [Figure 4]b, a significant reduction in the GSH-Px activity in the hippocampus was observed in the STZ group compared to the Sham group (P< 0.01). However, following the application of TEN (2, 4, and 8 mg/kg), the GSH-Px content was significantly higher in a dose-dependent manner than that of the STZ group (P< 0.05, P < 0.01).
The degree of free radical damage following STZ injection was assessed using lipid peroxidation, which was measured as the MDA level. As shown in [Figure 4]c, compared to the Sham group, MDA levels in the brain tissue significantly increased in the STZ group (P< 0.01); however, treatment with TEN (2, 4, and 8 mg/kg) significantly ameliorated the increases (P< 0.05, P < 0.01).
Effect of tenuigenin on 4-hydroxy-2-nonenal adducts levels
4-HNE is an α, β unsaturated aldehyde produced during oxidation in the membranes. 4-HNE can react with proteins to form 4-HNE adducts, and the levels of 4-HNE adducts in the hippocampus were also investigated as oxidative parameters. As shown in [Figure 5]a and [Figure 5]b, the hippocampal tissues of the STZ group had significantly higher 4-HNE levels compared to the Sham group (P< 0.01). Furthermore, 4-HNE in the rats treated with 2 mg/kg of TEN was lower than that of 4 and 8 mg/kg (P< 0.05, P < 0.01). The effects of 4 mg/kg and 8 mg/kg TEN on hippocampal 4-HNE were not significantly different.
|Figure 5: (a) Effect of tenuigenin treatment on 4-hydroxy-2-nonenal adducts. The relative values from each group were presented as mean ± standard deviation. Equal loading of proteins was illustrated by actin bands. (b) The level of 4-hydroxy-2-nonenal adducts was significantly increased in the intracerebroventricular - streptozotocin group as compared to the Sham group (##P < 0.01 vs. Sham group). Treatment with tenuigenin significantly restored these enhanced levels of 4-hydroxy-2-nonenal adducts as induced by intracerebroventricular - streptozotocin (**P < 0.01 vs. streptozotocin group, ◬P < 0.05 or ▴P < 0.01 vs. tenuigenin 2 mg/kg group)|
Click here to view
Effect of tenuigenin on phosphorylated tau proteins
We evaluated levels of tau phosphorylation and observed an increase at ser396 and thr181 sites in the hippocampus in ICV-STZ rats compared with the Sham group (P< 0.01). As illustrated in [Figure 6]a and [Figure 6]b, the total tau protein expression was not significantly changed among the groups (P > 0.05). Treatment with TEN (2, 4, and 8 mg/kg) obviously reversed the hyperphosphorylation of tau in the hippocampus of ICV-STZ rats [Figure 6]c and [Figure 6]d. In addition, treatment with the highest dose of TEN was more effective in decreasing the hyperphosphorylation of tau when compared to the effect of the medium and lowest TEN doses (P< 0.05, P < 0.01).
|Figure 6: (a) Effects of tenuigenin on intracerebroventricular - streptozotocin -induced phosphorylated tau. The level of ser396-phosphorylated tau and thr181-phosphorylated tau as induced by streptozotocin was significantly decreased after administration of tenuigenin. All values are expressed as mean ± standard deviation (n = 5). (##P < 0.01 vs. Sham group). **P < 0.01 versus streptozotocin group. ◬P < 0.05 or ▴P < 0.01 versus tenuigenin 2 mg/kg group. ★P < 0.05 versus tenuigenin 4 mg/kg group. (b) The level of total tau. (c) The level of ser396-phosphorylated tau. (d) The level of thr181-phosphorylated tau|
Click here to view
| Discussion|| |
AD is the most common cause of dementia characterized by the loss of memory and cognitive dysfunctions,, accounting for approximately 65%–70% of all dementia. AD is increasing rapidly as life expectancy is prolonged, and the disease has become a critical health-care issue and an important social problem worldwide.
ICV-STZ causes desensitization of insulin receptors, resulting in impaired insulin signaling.,, Our results today are consistent with previous reports ,, stating that ICV injection of STZ triggered an obvious impairment in memory and a broad range of neural changes including impaired metabolism and increases in oxidative stress, while administration of TEN prevented this cognitive damage.
TEN, a major active component of the Chinese herb Yuanzhi, has been described as having a wide range of activities on the pathological state of the organism by many studies.,,,, Recently, our laboratory has demonstrated that TEN has significant protective effects against the neurotoxicity induced by methylglyoxal in cultured hippocampal neural stem cells. Hence, the present study was designed to explore the neuroprotective effect of TEN on STZ-induced cognitive decline in a model of sporadic AD.
The MWM escape task was employed to assess spatial learning and memory abilities of animals by observing the escape latency to reach a hidden platform. In the present study, animals in the STZ group showed a slower rate of decline in the escape latency compared to the Sham group, whereas those treated with TEN showed shorter escape latencies in a dose-dependent manner. Furthermore, treatment with TEN showed a marked increase in the percentage of distance and time spent in the target quadrant. The MWM test suggests that TEN could protect rats from STZ-induced learning and memory impairment.
Previous reports have demonstrated that oxidative stress plays an important role in the advancement of ageing and age-related neurodegenerative disorders such as AD., Oxidative stress is the imbalance between the oxidative-antioxidative systems, which is a key mechanism involved in STZ-induced neurotoxicity. Free radical-induced damage to macromolecules such as lipid, protein, and nucleic acids is considered an important factor in the acceleration of AD.
MDA, an end-product of lipid hydroperoxide, is an indicator of ROS. SOD is a catalyzing enzyme that can reduce O2− to hydrogen peroxide (H2O2). GSH-Px protects cells by catalyzing the reduction of H2O2 and other peroxides. A decrease in the level of GSH-Px may inhibit H2O2 clearance and increase the formation of • OH.• OH is the most toxic molecule to the brain, resulting in more oxidant load and consequently oxidative damage. Too much H2O2 might accelerate the generation of the peroxidation of polyunsaturated fatty acids and lead to the formation of lipid peroxidation., Lipid peroxides and hydroperoxides can generate MDA and HNE, which are more stable and diffusible cytotoxic agents. The brain is a main metabolizer organ of oxygen and seems to be more susceptible to oxidative stress.
In the present study, the increased MDA level in the ICV-STZ animals was attenuated by TEN treatment. TEN could also increase the SOD and GSH-Px activities in the hippocampus of STZ-treated rats. Previous studies have revealed that TEN has antioxidative properties and can protect against oxidative stress in vitro. In this work, we guess that ameliorating oxidative stress may be one of the reasons TEN improving learning and memory abilities. Our previous study suggested that TEN was involved in regulating the proliferation and differentiation of hippocampal neural stem cells in vitro. However, whether there is similar effect of TEN on hippocampal neuronsin vivo not yet known and we hope that our today's work can clarify the potential therapeutic value of TEN in the treatment of cognitive dysfunction.
4-HNE is a kind of aldehyde produced during lipid peroxidation of unsaturated lipids in the membrane. It can conjugate to specific amino acid residues of proteins and play key roles in signal transduction pathways, cellular adhesion, and numerous cell cycle events. The previous report suggested that there exist increased levels of specific HNE-histidine Michael adducts in AD hippocampus compared to age-matched controls. The results of the present study are similar to the previous report demonstrating that HNE was significantly increased in ICV-STZ rats. Moreover, after TEN treatment, the HNE level decreased markedly.
Tau protein is a kind of microtubule-associated protein, and the main function of tau is to maintain intracellular microtubule stabilization. Hyperphosphorylation of tau decreases the capacity for binding and stabilizing microtubules. Phosphorylated tau protein aggregates to neurofibrillary tangles (NFTs), which are one of the typical characteristics of the hallmark pathologies of AD. Phosphorylation of tau proteins on many sites is toxic to neurons and inhibits normal tubulin, leading to axonal transport dysfunction, proteasome activity inhibition, consequent impairment of the structure and function of neurons, and eventually neurodegeneration.
In the present study, we also investigated the tau hyperphosphorylation expression induced by ICV-STZ. Previous reports showed that the accumulation of HNE produced hyperphosphorylated tau proteins; furthermore, under conditions of modified HNE, normal tau proteins are vulnerable to be hyperphosphorylated, and epitopes of these modified tau represent the major conformational changes contributing to the formation of NFTs. In this study, we found that increased 4-HNE was accompanied by elevation of phosphorylated tau at ser396 and thr181 in ICV-STZ rats. After TEN administration, 4-HNE decreased and TEN could reduce hyperphosphorylation of tau protein at these two sites, suggesting the role of TEN in the suppression of p-tau activation and associated pathogenesis caused by STZ.
| Conclusion|| |
The present study shows that TEN can ameliorate behavioral impairments of memory deficit, inhibit oxidative stress, and block tau hyperphosphorylation in rat hippocampus induced by the administration of ICV-STZ. Taken together, these data suggest that TEN may be a useful agent for the treatment of AD.
Financial support and sponsorship
This work was supported by A Project Funded by the Youth Programme of Beijing Municipal Administration of Hospitals (grant no. QML20160805) and Beijing Municipal Administration of Hospitals Incubating Program (code: PZ2016027). The assistance of the staff is gratefully acknowledged.
Conflicts of interest
There are no conflicts of interest.
| References|| |
Firuzi O, Praticò D. Coxibs and Alzheimer's disease: Should they stay or should they go? Ann Neurol 2006;59:219-28.
Aliev G, Priyadarshini M, Reddy VP, Grieg NH, Kaminsky Y, Cacabelos R, et al.
Oxidative stress mediated mitochondrial and vascular lesions as markers in the pathogenesis of Alzheimer disease. Curr Med Chem 2014;21:2208-17.
Giraldo E, Lloret A, Fuchsberger T, Viña J. Aβ and tau toxicities in Alzheimer's are linked via oxidative stress-induced p38 activation: Protective role of vitamin E. Redox Biol 2014;2:873-7.
Melov S, Adlard PA, Morten K, Johnson F, Golden TR, Hinerfeld D, et al.
Mitochondrial oxidative stress causes hyperphosphorylation of tau. PLoS One 2007;2:e536.
Zhou S, Yu G, Chi L, Zhu J, Zhang W, Zhang Y, et al.
Neuroprotective effects of edaravone on cognitive deficit, oxidative stress and tau hyperphosphorylation induced by intracerebroventricular streptozotocin in rats. Neurotoxicology 2013;38:136-45.
Salkovic-Petrisic M, Osmanovic-Barilar J, Brückner MK, Hoyer S, Arendt T, Riederer P, et al.
Cerebral amyloid angiopathy in streptozotocin rat model of sporadic Alzheimer's disease: A long-term follow up study. J Neural Transm (Vienna) 2011;118:765-72.
Shoham S, Bejar C, Kovalev E, Schorer-Apelbaum D, Weinstock M. Ladostigil prevents gliosis, oxidative-nitrative stress and memory deficits induced by intracerebroventricular injection of streptozotocin in rats. Neuropharmacology 2007;52:836-43.
Lester-Coll N, Rivera EJ, Soscia SJ, Doiron K, Wands JR, de la Monte SM, et al.
Intracerebral streptozotocin model of type 3 diabetes: Relevance to sporadic Alzheimer's disease. J Alzheimers Dis 2006;9:13-33.
Grünblatt E, Salkovic-Petrisic M, Osmanovic J, Riederer P, Hoyer S. Brain insulin system dysfunction in streptozotocin intracerebroventricularly treated rats generates hyperphosphorylated tau protein. J Neurochem 2007;101:757-70.
Salkovic-Petrisic M, Hoyer S. Central insulin resistance as a trigger for sporadic Alzheimer-like pathology: An experimental approach. J Neural Transm Suppl 2007;72:217-33.
Yuan HL, Li B, Xu J, Wang Y, He Y, Zheng Y, et al.
Tenuigenin protects dopaminergic neurons from inflammation-mediated damage induced by the lipopolysaccharide. CNS Neurosci Ther 2012;18:584-90.
Zhang H, Han T, Zhang L, Yu CH, Wan DG, Rahman K, et al.
Effects of tenuifolin extracted from radix polygalae on learning and memory: A behavioral and biochemical study on aged and amnesic mice. Phytomedicine 2008;15:587-94.
Jia H, Jiang Y, Ruan Y, Zhang Y, Ma X, Zhang J, et al.
Tenuigenin treatment decreases secretion of the Alzheimer's disease amyloid beta-protein in cultured cells. Neurosci Lett 2004;367:123-8.
Chen Q, Li LK. Protective effect of tenuigenin on cytotoxicity of primary cultures of cortical neurons induced by amyloid beta-protein 1-40 (Abeta (1-40)). Zhongguo Zhong Yao Za Zhi 2007;32:1336-9.
Chen Y, Huang X, Chen W, Wang N, Li L. Tenuigenin promotes proliferation and differentiation of hippocampal neural stem cells. Neurochem Res 2012;37:771-7.
Chen YJ, Huang XB, Li ZX, Yin LL, Chen WQ, Li L, et al.
Tenuigenin protects cultured hippocampal neurons against methylglyoxal-induced neurotoxicity. Eur J Pharmacol 2010;645:1-8.
Sharma M, Gupta YK. Effect of alpha lipoic acid on intracerebroventricular streptozotocin model of cognitive impairment in rats. Eur Neuropsychopharmacol 2003;13:241-7.
Gandy S. Molecular basis for anti-amyloid therapy in the prevention and treatment of Alzheimer's disease. Neurobiol Aging 2002;23:1009-16.
Koo JH, Kwon IS, Kang EB, Lee CK, Lee NH, Kwon MG, et al.
Neuroprotective effects of treadmill exercise on BDNF and PI3-K/Akt signaling pathway in the cortex of transgenic mice model of Alzheimer's disease. J Exerc Nutrition Biochem 2013;17:151-60.
Blennow K, de Leon MJ, Zetterberg H. Alzheimer's disease. Lancet 2006;368:387-403.
Brookmeyer R, Johnson E, Ziegler-Graham K, Arrighi HM. Forecasting the global burden of Alzheimer's disease. Alzheimers Dement 2007;3:186-91.
Hoyer S. The aging brain. Changes in the neuronal insulin/insulin receptor signal transduction cascade trigger late-onset sporadic Alzheimer disease (SAD). A mini-review. J Neural Transm (Vienna) 2002;109:991-1002.
Hoyer S. The brain insulin signal transduction system and sporadic (type II) Alzheimer disease: An update. J Neural Transm (Vienna) 2002;109:341-60.
Hoyer S, Müller D, Plaschke K. Desensitization of brain insulin receptor. Effect on glucose/energy and related metabolism. J Neural Transm Suppl 1994;44:259-68.
Sonkusare S, Srinivasan K, Kaul C, Ramarao P. Effect of donepezil and lercanidipine on memory impairment induced by intracerebroventricular streptozotocin in rats. Life Sci 2005;77:1-4.
Lannert H, Hoyer S. Intracerebroventricular administration of streptozotocin causes long-term diminutions in learning and memory abilities and in cerebral energy metabolism in adult rats. Behav Neurosci 1998;112:1199-208.
Sharma M, Gupta YK. Intracerebroventricular injection of streptozotocin in rats produces both oxidative stress in the brain and cognitive impairment. Life Sci 2001;68:1021-9.
Jiang T, Sun Q, Chen S. Oxidative stress: A major pathogenesis and potential therapeutic target of antioxidative agents in Parkinson's disease and Alzheimer's disease. Prog Neurobiol 2016;147:1-9.
Chong ZZ, Li F, Maiese K. Oxidative stress in the brain: Novel cellular targets that govern survival during neurodegenerative disease. Prog Neurobiol 2005;75:207-46.
Liu R, Liu IY, Bi X, Thompson RF, Doctrow SR, Malfroy B, et al.
Reversal of age-related learning deficits and brain oxidative stress in mice with superoxide dismutase/catalase mimetics. Proc Natl Acad Sci U S A 2003;100:8526-31.
Cao YW, Jiang Y, Zhang DY, Wang M, Chen WS, Su H, et al.
Protective effects of Penthorum
Chinense Pursh against chronic ethanol-induced liver injury in mice. J Ethnopharmacol 2015;161:92-8.
Dringen R, Gutterer JM, Hirrlinger J. Glutathione metabolism in brain metabolic interaction between astrocytes and neurons in the defense against reactive oxygen species. Eur J Biochem 2000;267:4912-6.
Sun Y. Free radicals, antioxidant enzymes, and carcinogenesis. Free Radic Biol Med 1990;8:583-99.
Dringen R. Metabolism and functions of glutathione in brain. Prog Neurobiol 2000;62:649-71.
Sun GB, Deng XC, Li CH. The protective effects of tenuigenin on the PC12 cells injury induced by H2O2. Zhong Yao Cai 2007;30:991-3.
Fukuda M, Kanou F, Shimada N, Sawabe M, Saito Y, Murayama S, et al.
Elevated levels of 4-hydroxynonenal-histidine michael adduct in the hippocampi of patients with Alzheimer's disease. Biomed Res 2009;30:227-33.
Hanger DP, Anderton BH, Noble W. Tau phosphorylation: The therapeutic challenge for neurodegenerative disease. Trends Mol Med 2009;15:112-9.
Riley KP, Snowdon DA, Markesbery WR. Alzheimer's neurofibrillary pathology and the spectrum of cognitive function: Findings from the Nun Study. Ann Neurol 2002;51:567-77.
Alonso Adel C, Mederlyova A, Novak M, Grundke-Iqbal I, Iqbal K. Promotion of hyperphosphorylation by frontotemporal dementia tau mutations. J Biol Chem 2004;279:34873-81.
Yang Y, Yang XF, Wang YP, Tian Q, Wang XC, Li HL, et al.
Inhibition of protein phosphatases induces transport deficits and axonopathy. J Neurochem 2007;102:878-86.
Ren QG, Liao XM, Chen XQ, Liu GP, Wang JZ. Effects of tau phosphorylation on proteasome activity. FEBS Lett 2007;581:1521-8.
Liu Q, Smith MA, Avilá J, DeBernardis J, Kansal M, Takeda A, et al.
Alzheimer-specific epitopes of tau represent lipid peroxidation-induced conformations. Free Radic Biol Med 2005;38:746-54.
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6]
|This article has been cited by|
||Discovery of a Novel Lineage Burkholderia cepacia ST 1870 Endophytically Isolated from Medicinal Polygala paniculata Which Shows Potent In Vitro Antileishmanial and Antimicrobial Effects
| ||Felipe de Paula Nogueira Cruz,Ailton Ferreira de Paula,Camila Tita Nogueira,Paulo Henrique Marques de Andrade,Leonardo Maurici Borges,Paulo Teixeira Lacava,Ilana Lopes Baratella da Cunha Camargo,Fernanda de Freitas Aníbal,Cristina Paiva de Sousa,Simona Nardoni |
| ||International Journal of Microbiology. 2021; 2021: 1 |
|[Pubmed] | [DOI]|
||The Effects of Yuan-Zhi Decoction and Its Active Ingredients in Both In Vivo and In Vitro Models of Chronic Cerebral Hypoperfusion by Regulating the Levels of Aß and Autophagy
| ||Yan Liu,Xiaobo Huang,Wenqiang Chen,Yujing Chen,Ningqun Wang,Xiling Wu |
| ||Evidence-Based Complementary and Alternative Medicine. 2020; 2020: 1 |
|[Pubmed] | [DOI]|
||Polygala tenuifolia: a source for anti-Alzheimer’s disease drugs
| ||Xinxin Deng,Shipeng Zhao,Xinqi Liu,Lu Han,Ruizhou Wang,Huifeng Hao,Yanna Jiao,Shuyan Han,Changcai Bai |
| ||Pharmaceutical Biology. 2020; 58(1): 410 |
|[Pubmed] | [DOI]|
||Investigation of Low Dose Cabazitaxel Potential as Microtubule Stabilizer in Experimental Model of Alzheimeræs Disease: Restoring Neuronal Cytoskeleton
| ||Pallavi Duggal,Kuldeep S. Jadaun,Ehraz M. Siqqiqui,Sidharth Mehan |
| ||Current Alzheimer Research. 2020; 17(7): 601 |
|[Pubmed] | [DOI]|
||Apelin-13 Suppresses Neuroinflammation Against Cognitive Deficit in a Streptozotocin-Induced Rat Model of Alzheimer’s Disease Through Activation of BDNF-TrkB Signaling Pathway
| ||Huaiqing Luo,Yang Xiang,Xiangping Qu,Huijun Liu,Chi Liu,Guangyi Li,Li Han,Xiaoqun Qin |
| ||Frontiers in Pharmacology. 2019; 10 |
|[Pubmed] | [DOI]|
||Effect of a-asarone on ethanol-induced learning and memory impairment in mice and its underlying mechanism
| ||Qinqing Li,Feifei Xu,Qi Zhang,Xiaoqing Li,Meitong Guo,Yiming Zhang,Zilin Wang,Jie Wang,Jiaqi Zhao,Yajuan Tian,Zhao Zhang,Shifeng Chu,Wenbin He |
| ||Life Sciences. 2019; : 116898 |
|[Pubmed] | [DOI]|
||Design and synthesis of pentacyclic triterpene conjugates and their use in medicinal research
| ||Jiri Hodon,Lucie Borkova,Jan Pokorny,Anna Kazakova,Milan Urban |
| ||European Journal of Medicinal Chemistry. 2019; : 111653 |
|[Pubmed] | [DOI]|
||A review on the phytopharmacological studies of the genus Polygala
| ||Marie-Aleth Lacaille-Dubois,Clément Delaude,Anne-Claire Mitaine-Offer |
| ||Journal of Ethnopharmacology. 2019; : 112417 |
|[Pubmed] | [DOI]|