Research progress on the use of Rhizoma coptidis extract against Alzheimer's disease / 中华老年医学杂志

Rhizoma coptidis extract has a variety of pharmacological activities, including alleviating cognitive impairment in Alzheimer's disease(AD). The main mechanisms of its anti-AD activity include reducing the production of amyloid β(Aβ), inhibiting the phosphorylation of Tau protein, inhibiting cholinesterase, anti-inflammation, anti-oxidation, improving apoptosis, etc.This paper reviewed the anti-AD effect of Rhizoma coptidis extract and the specific mechanisms, aiming to provide a theoretical basis for relevant research and clinical practice.

Alterations of gut microbiota composition and cogni-tive behavior in APP/PS1/tau triple-transgenic mouse model of Alzheimer's disease / 中国药理学与毒理学杂志

OBJECTIVE Alzheimer's disease(AD)is a progressive neurological disease.Given the important role of gut microbiota composition in AD pathology,the observed perturbation in the microbiota composition and diversity may serve as the mechanisms underlying age-dependent APP/PS1/tau triple-transgenic mouse(3×Tg-AD)mice amyloid deposition and memory deficits.Here-in,we intended to investigate the gut microbiota and as-sessed its relationship with the triggering and develop-ment of cognitive impairment of AD.METHODS This study involves the comparative assessment of spatial learning,amyloid β-protein(Aβ)accumulation,and fecal microbiota alterations in 3×Tg-AD mice from three age groups:AD asymptomatic stage(3 m),presymptomatic stage(6 m),and the symptomatic stage of AD(9 m).RE-SULTS We demonstrate that spatial memory deficits,brain Aβ accumulation,and weight gain in 3×Tg-AD mice gradually appear after 6 months of age.However,the total gut bacterial counts underwent changes from 3 to 6 months of age and were further altered at 9 months of age.Importantly,changes in gut bacteria abundance of Desulfobacterota and Actinobacteriota phylain 6-month-old mice preceded apparent spatial memory deficits.CONCLUSION Changes in the gut microbial community are one of the mechanisms of early AD pathology.

Amelioration of Cognitive Dysfunction in Diabetic Rats by Jianpi Qinghua Prescription / 中国实验方剂学杂志

ObjectiveTo explore the amelioration of cognitive dysfunction in diabetes mellitus （DM） by Jianpi Qinghua prescription （JPQH） based on type 2 diabetes （T2DM） model rats. MethodFifty healthy male Wistar rats of SPF grade were randomly divided into control group （n=10） and experimental group （n=40）. The rats in the control group were fed conventionally， while those in the experimental group were fed on a high-sugar， high-fat diet for six weeks and administered with streptozotocin （STZ） for the induction of the DM model. The model rats were randomly divided into model group， sitagliptin group （1.2 g·L-1）， pioglitazone group （0.8 g·L-1）， and JPQH group （1.3 g·mL-1）， with 10 rats in each group. After six weeks of drug intervention， the changes in body weight， blood glucose， and other related indexes of each group were recorded. Enzyme-linked immunosorbent assay （ELISA） was performed to detect the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） in the peripheral blood and brain. The Morris water maze test was used to evaluate the cognitive function in rats. Hematoxylin-eosin （HE） staining was used to observe the pathological morphology of the hippocampal CA region. The amyloid β-protein 40 （Aβ40） level was detected by immunohistochemistry. The protein expression of t-tau and p-tau in hippocampal neurons of rats was detected by Western blot. ResultCompared with blank group， the body weight of model group was significantly decreased （P<0.05）， blood glucose level was significantly increased （P<0.01）， inflammatory cytokines TNF-α and IL-1β were increased （P<0.05）， learning and spatial ability were significantly decreased （P<0.01）， the arrangement of hippocampal cells was loose and disordered， and the intercellular space was significantly increased. The number of cells decreased significantly， and the expression of Aβ40 increased significantly. and increased t-tau and p-tau protein content in the hippocampus （P<0.01）. Compared with model group， the JPQH group showed reduced blood glucose （P<0.01）， decreased TNF-α and IL-1β levels in the peripheral blood and cerebrospinal fluid （P<0.05）， a downward trend of IL-6 without a statistical difference， improved learning and spatial memory ability （P<0.01）， densely arranged cells in the hippocampal CA1 area， increased cell number， reduced Aβ40 expression， and decreased p-tau protein expression （P<0.05）. ConclusionJPQH can prevent cognitive dysfunction in DM by reducing inflammatory factor levels， decreasing neurotoxicity caused by Aβ40 deposition， and inhibiting hyperphosphorylation of tau protein in DM rats.

Research progress in clinical treatment of Alzheimer's disease and potential drugs from natural products / 药学学报

With the acceleration of the aging process of our country's population, the impact of aging-related diseases - Alzheimer's disease (AD) on society and families has become increasingly prominent. AD is caused by multiple mechanisms, and the pathogenesis has not been fully elucidated. Most of the clinical treatments are single therapy, which mainly focuses on improving symptoms and are difficult to reverse the disease process. Therefore, the development of drugs that can both improve symptoms and reverse the disease process is extremely urgent in clinical. Increasing number of studies has shown that traditional Chinese medicine plays an important role in the prevention and treatment of AD. The natural products have many advantages, such as novel structures, multiple targets and diverse activities, which can be used as an important source of leading compounds for the treatment of AD. The review summarizes the main clinical treatment methods and the research progress of natural ingredients in traditional Chinese medicine, and provides a reference for the follow-up clinical treatment of AD combined with the advantages of traditional Chinese medicine treatment.

Ginsenoside Rg_1 protects PC12 cells against Aβ-induced injury through promotion of mitophagy by PINK1/parkin activation / 中国中药杂志

Amyloid β-protein(Aβ) deposition in the brain is directly responsible for neuronal mitochondrial damage of Alzheimer's disease(AD) patients. Mitophagy, which removes damaged mitochondria, is a vital mode of neuron protection. Ginsenoside Rg_1(Rg_1), with neuroprotective effect, has displayed promising potential for AD treatment. However, the mechanism underlying the neuroprotective effect of Rg_1 has not been fully elucidated. The present study investigated the effects of ginsenoside Rg_(1 )on the autophagy of PC12 cells injured by Aβ_(25-35) to gain insight into the neuroprotective mechanism of Rg_1. The autophagy inducer rapamycin and the autophagy inhi-bitor chloroquine were used to verify the correlation between the neuroprotective effect of Rg_1 and autophagy. The results showed that Rg_1 enhanced the viability and increased the mitochondrial membrane potential of Aβ-injured PC12 cells, while these changes were blocked by chloroquine. Furthermore, Rg_(1 )treatment increased the LC3Ⅱ/Ⅰ protein ratio, promoted the depletion of p62 protein, up-regulated the protein levels of PINK1 and parkin, and reduced the amount of autophagy adaptor OPTN, which indicated the enhancement of autophagy. After the silencing of PINK1, a key regulatory site of mitophagy, Rg_1 could not increase the expression of PINK1 and parkin or the amount of NDP52, whereas it can still increase the LC3Ⅱ/Ⅰ protein ratio and promote the depletion of OPTN protein which indicated the enhancement of autophagy. Collectively, the results of this study imply that Rg_1 can promote autophagy of PC12 cells injured by Aβ, and may reduce Aβ-induced mitochondrial damage by promoting PINK1-dependent mitophagy, which may be one of the key mechanisms of its neuroprotective effect.

Effects of on behavior and blood-brain barrier in Alzheimer's disease mice / 浙江大学学报·医学版

To investigate the effects of on behavior and blood brain barrier (BBB) in Alzheimer's disease mice. Thirty-eight 4-month-old APP/PS1 double transgenic mice were randomly divided into three groups: model group, low-dose group and high-dose group. Saline, and 12 g·kg·d were given to each group by continuous gavage once a day for respectively. The changes in activities of daily live and fear conditioning memory behavior of mice were examined by nesting behavior test and fear conditioning test, respectively. The β-amyloid protein (Aβ) depositions in cortex and hippocampal CA1 area of mice were detected by thioflavin T staining. The CD34 and activities fibrinogen (Fib) immunofluorescence double staining were used to determine the vascular endothelial integrity and BBB exudation. Compared with model mice, activities of daily live were significantly improved in low-dose and high-dose groups (both <0.01), the fear memory ability was significantly increased in high-dose group (<0.01). The amount of Aβ deposition in cortex and hippocampal CA1 decreased significantly in high-dose group, the area ratio decreased significantly; the area ratio of Aβ deposition in hippocampal CA1 region in low-dose group also decreased (all <0.05). The proportions of CD34 positive area of cortex in low and high dose groups increased, the percentage of fibrinogen positive area decreased (all <0.05). The proportion of CD34 positive area in hippocampal CA1 region in high-dose group was significantly increased, the percentage of fibrinogen positive area decreased significantly (both <0.05). especially high-dose can improve the activities of daily live and fear conditioning memory function of APP/PS1 mice, reduce the deposition of Aβ in brain. The mechanism may be related to the reduction of BBB permeability and the protection of the integrity of BBB.

Protective effect of baicalein on injured PC12 cell induced by Aβ and its mechanism / 国际中医中药杂志

Objective:To investigate the protective effect of baicalein on injured PC12 cell induced by Aβ and explore its mechanism.Methods:The method of MTT was used to detect the cell activity of each group and screened the concentration of baicalein. The PC12 cells were randomly divided into the blank group, the Aβ group, the baicalin group and the estradiol group. 24 hours after inoculation, baicalein group was intervened with 1×10 -6 mol/L baicalein solution, and estradiol group was intervened with 1×10 -5 mol/L estradiol solution. Two hours later, except the blank group, the other groups were added with 1.5×10 -4 mol/L Aβ to make the model. MTT assay was used to detect the cell viability of each group after 24 hours of cultivation. Then used oxidation kit to detect the contents of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and lactate dehydrogenase (LDH) in each group. And the level of caspase-3 mRNA was detected by Real-time Quantitative PCR (RT-PCR). Then the Western blot method was used to detect the expressions of p-PI3K, p-AKT and caspase-3. Results:Compared with the Aβ group, the PC12 cell viability [(96.348 ± 0.571)%, (97.183 ± 0.714)% vs. (86.922 ± 0.429)%] in the baicalin group and the estradiol group significantly increased( P<0.01). The activities of SOD [(54.31 ± 1.34) U/mgprot, (57.38 ± 2.25) U/mgprot vs. (36.18 ± 2.24) U/mgprot] and GSH-PX [(4.46 ± 0.23) U/mgprot, (4.72 ± 0.31) U/mgprot vs. (2.05 ± 0.37) U/mgprot] significantly increased, and the level of LDH [(85.43 ± 0.92) nmol/ml, (82.46 ± 0.27) nmol/ml vs. (99.17 ± 0.52) nmol/ml] significantly decreased ( P<0.01). The expression of caspase-3 mRNA (2.24 ± 0.64, 2.33 ± 0.75 vs. 3.46 ± 0.46) and p-PI3K (0.46 ± 0.03, 0.44 ± 0.06 vs. 0.66 ± 0.09), p-AKT (0.43 ± 0.05, 0.41 ± 0.02 vs. 0.58 ± 0.03), caspase-3 (0.61 ± 0.03, 0.56 ± 0.53 vs. 0.92 ± 0.07) protein significantly decreased ( P<0.01). Conclusion:Baicalein could slow down cell apoptosis and oxidative reaction, reduce the damage of Aβ to PC12 cells by inhibiting the expression of PI3K/AKT pathway.

Tolcapone derivative PCDNA inhibits Aβ42 fibrillogenesis and reduces its cytotoxicity / 药学学报

Abnormal aggregation of amyloid-β protein (Aβ) in brain plays a vital role in the occurrence of Alzheimer's disease (AD). Hence, inhibiting Aβ aggregation is one major tactic for therapy of AD. Previous studies have found that tolcapone can inhibit Aβ42 aggregation and reduce the cytotoxicity induced by Aβ42 aggregates, but clinical studies have found that tolcapone has strong liver toxicity. To reduce the liver toxicity of tolcapone, its side chain structure was modified to obtain its derivative phenethyl (E)-2-cyano-3-(3,4 dihydroxy-5-nitrobenzene)-acrylate (PCDNA). Thioflavin T (ThT) and atomic force microscopy (AFM) assays were used to explore the inhibitory effect of PCDNA on Aβ42 fibrillogenesis. The cytotoxicity assays were used to explore the inhibitory effect of PCDNA against the cytotoxicity induced by Aβ42 aggregates. In addition, the depolymerization effect of PCDNA on mature Aβ42 fibrils was also explored. Finally, molecular docking was used to explore the interaction between PCDNA and Aβ42 pentamer. These results lay the foundation for the study of the structural analogues of tolcapone as Aβ inhibitors.

Nucleotide-binding Oligomerization Domain-like Receptor Family Pyrin Domain Containing 3 Inflammasome in Alzheimer's Disease / 中国医学科学院学报

Alzheimer's disease(AD)is a chronic neurodegenerative disease whose cause remains unclear.The β-amyloid plaques in the brain are one of the major pathological features of AD.However,the drugs targeting extracellular β-amyloid plaques have failed to cure the disease.Innate immunity and neuroinflammation play a role in the pathogenesis and progression of AD.As the macrophages existing in the central nervous system,microglia are related with extracellular β-amyloid deposition,intracellular neurofibrillary tangle formation,and neuron injury.Accumulating evidence demonstrates that the activation of nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3(NLRP3)inflammasome in microglia plays a role in AD,suggesting new therapeutic target for AD in this signaling pathway.This article reviewed the studies about the activation and regulation of NLRP3 inflammasome in the pathogenesis and progression of AD as well as the development of AD therapies targeting this pathway,aiming to provide reference for further studies in this field.

Effects of Effective Part of Tinospora sinensis on Hippocampus Proteomics of AD Rats Induced by Beta-amyloid Protein and D-galactose / 中国药学杂志

OBJECTIVE: To investigate the effect of the 80% ethanol elution part of Tinospora sinensis macroporous resin extract on the expression of hippocampus proteome in Alzheimer′s disease (AD) model rats induced by D-galactose combined with Aβ2535. METHODS: The AD rats model replicated by D-galactose combined with Aβ2535, The AD rat model was replicated by D-galactose combined with Aβ2535, and randomly divided into sham operation group, model group, Donepezil group (donepezil, 6.0 mg•kg-1), and 80% extraction of Tinospora sinensi group (crude drug 6 g•kg-1). Donepezil group: donepezil 0.1 mL•10 g-1 ig. 80% extraction of Tinospora sinensi group: Tinospora sinensis effective part extraction 0.1 mL•10 g-1 ig. Model group and sham-operation group: physiological saline 0.1 mL•10 g-1 ig. Once a day, continuous administration for 15 d. Separating the hippocampus and extracting the protein, take the system test with nanol-ESI liquid-mass spectrometry, protein discovery software was used for identification, and qualitative analysis different groups of hippocampal proteins by SIEVE software. Take the GO analysis on differential protein with the ANTHER classification system and use IPAD to enrich the pathway. RESULTS: Compared with the model group, the drug-administered group had 66 differential proteins, including tubulin, heat shock proteins, energy metabolism-related proteins, vesicle production/transport related proteins, and brain protection-related proteins, which are closely related to AD. The above differential proteins involve a total of 21 signaling pathways. CONCLUSION: Tinospora sinensis may promote the synthesis and release of neurotransmitters by up-regulating clathrin and vesicle-forming transport and neurotransmitter release, and improve the function of cholinergic function in the brain to achieve the pathological process of AD.

Progress in interventional mechanism of paeoniflorin on Alzheimer’s disease / 中草药

Alzheimer’s disease (AD) is one of the main pathogenies of the cognitive decline in aging population. Paeoniflorin is a water-soluble monoterpene isolated from the roots of Paeonia lactiflora or P. suffruticosa, which are both in the family Ranunculaceae. Paeoniflorin has the effect of improving the cognitive ability, sedative, anti-inflammatory, analgesic, and spasmolytisch activities. It could also regulate hyperphosphorylation of Tau and Aβ proteins; Paeoniflorin can restore the balance of neurotransmitter and neurotransmitter receptors including acetylcholine, nerve growth factor, calcium ion, α7 nicotinic acetylcholine receptor and adenosine receptors, and reduce inflammation, oxidative stress and neuron apoptosis in AD animal and cell models. This paper reviewed relevant research in the past decade about the effect of paeoniflorin on AD, which may provide theoretical basis to use agent from Chinese medicine for AD prevention and treatment.

Effect and its mechanism of Linagliptin on mild cognitive impairment in elderly type 2 diabetes mellitus patients / 中华老年医学杂志

Objective To investigate the effect and its underlying mechanism of Linagliptin on mild cognitive impairment (MCI) in elderly type 2 diabetes mellitus (T2DM) patients.Methods Montreal Cognitive Assessment(MoCA)scale was used to prospectively screen T2DM patients for MCI in our hospital from December 2016 to June 2017,and a total of 98 elderly T2DM patients with MCI were recruited.They were randomly divided into the linagliptin group(Linagliptin + metformin,n=50)and the non-linagliptin group(gliclazide + metformin,n =48).Serum fasting plasma glucose (FPG),glycosylated hemoglobin(HbAlc),blood lipids and amyloid β-protein 1-42 (Aβ1-42) levels were determined,and MoCA score and homeostasis model assessment of insulin resistance(HOMA-IR)were calculated,and were compared between the two groups before and after 24 weeks of treatment.Results In the linagliptin group,serum FPG,HbA1c,HOMA-IR,Aβ1-42 levels were significantly decreased and MoCA score was increased after 24 weeks of treatment as compared with pre-treatment [(7.29± 1.00) mmol/L vs.(9.16 ± 1.60) mmol/L,(7.19 ± 0.99) ％ vs.(9.36 ± 1.07) ％,(3.05 ± 1.12) vs.(4.05±1.30),(0.463±0.093)g/L vs.(0.528±0.110)g/L,(24.48± 1.18) vs.(23.22± 1.37),all P＜0.05].In the non-linagliptin group as control,FPG and HbA1c levels were decreased after 24 weeks of treatment as compared with pre-treatment[(7.23±1.09)mmol/L vs.(9.20± 1.75) mmol/L,(7.23±1.03)％ vs.(9.69± 1.18)％,both P ＜ 0.05],while there was no significant difference in HOMA IR,Aβ1-42 level and MoCA score[(3.95 ± 1.00) vs.(4.19± 1.13),(0.517± 0.113)g/L vs.(0.526±0.119)g/L,(23.21±1.18) vs.(23.00±1.32),all P＞0.05].It is worth to pay close attention to the key discovery of this paper that HOMA-IR and Aβ1-42 levels were significantly lower and MoCA score was significantly higher in the linagliptin group than in the non-linagliptin group after 24 weeks of treatment(all P＜0.05).Conclusions Linagliptin as one of DPP-4 enzyme inhibitors can improve the cognitive function in elderly patients with T2DM,which might be relevant to reducing serum Aβ level and improving HOMA-IR.DPP-4 enzyme inhibitor may be a good option for treatment of mild cognitive dysfunction in T2DM patients in the future.

Effects of Chinese traditional medicine-selected recipe Q0409 on ability of learning and memory in SAM-P/8 mice / 中国病理生理杂志

AIM:To investigate the effects of Chinese traditional medicine-selected recipe Q0409 on the ability of learning and memory in SAM-P/8 mice. METHODS:Total 91 mice (4-month-old SAM-P/8 mice, SAM-R/1 mice and Kunming mice) were used in the study, in which the male and female animals were labeled separately. According to the performance of Morris water maze test, the mice were divided into 5 groups randomly. The mice were fed with different drugs or distilled water for 60 d (from 4 months to 6 months). The mice were fed with the drugs from 61 d to 65 d, and 1 h later each time, the Morris water maze test was carried out. After this Morris test were finished at 65 d, the mice were killed immediately and their hippocampal tissues were isolated. Half of the hippocampal tissues were added with precooled normal saline and made into 10% (g/mL) homogenate for detecting the protein content and acetyl cholinesterase (AChE) activity. The other half was fixed with 4% paraformaldehyde and embedded with paraffin for immunohistochemical staining of amyloid β-protein (Aβ). RESULTS:Compared with model group, the results of navigation training and spatial probe training in Morris water maze test were significantly improved (P<0.05), and the activity of AChE in the hippocampal ho-mogenate was significantly decreased (P<0.05) in Q0409 treatment group. No difference in Q0409 group was observed compared with control group and positive drug (huperzine A) group. Immunohistochemical staining showed no typical "se-nile plaques" in the male mice of Q0409 group, while there was shallower and smaller brown staining in the hippocampus of the female mice of Q0409 group. The positive area of Aβ deposition decreased in the CA1 area of hippocampal tissues in Q0409 group. These results were similar to those in positive drug group. CONCLUSION:Q0409 improves the ability of learning and memory in SAM-P/8 mice, which is related to the inhibition of AChE activity and the reduction of Aβ protein deposition in the hippocampus. The effects is similar to those of huperzine A.

Changes in histopathology and ultrastructure of corneal epithelium in TgAPPswePS1 transgenic mice / 眼科新进展

Objective To investigate the histopathological and ultrastructural changes in the corneal epithelium in TgAPPswePS1 transgenic mice.Methods TgAPPswePS1 transgenic mice were randomly divided into experimental group (A and B sub-group) and control group.There were 15 APPswe/PSEN1dE9 transgenic mice (15-18 months old) with Alzheimer's disease (AD) in the experimental A group,and 15 APPswe/PSEN1 dE9 transgenic AD mice (8 months old) mice in the experimental B group,as well as 10 wild-type mice (8 months old) in the control group.Then,the histopathological and ultrastructural changes and the expression of amyloid β-protein (Aβ) in the corneal epithelium of the mice were detected,and finally,the apoptosis of corneal epithelial cells were observed by TUNEL assay.Results The thickness of corneal epithelium in the control group,A and B sub-group of the experimental group was (23.567 ± 2.123) μm,(15.456 ± 1.439) μm and (20.104 ± 1.763) μm,respectively.Meanwhile,murine corneal epithelial cells presented the histopathological changes of disorderly arrangement,decreased layers of cells and irregular morphology in the experimental group compared with the control group.Under transmission electron microscope,the microvilli on the surface of corneal epithelium was flat and significantly decreased in the A and B sub-group when compared with the control group.Moreover,Aβ positive expression in the experimental group was significantly upregulated compared with the control group;and the apoptotic number of the corneal epithelium in the B sub-group [(5.631 ± 2.471) cells] was smaller than that in the A sub-group [(16.329 ±3.542) cells],with the significant difference(P ＜ 0.05).Conclusion There are changes in the histopathology and ultrastructure of the corneal epithelium and the expression of Aβ in TgAPPswePS1 transgenic mice compared with wild-type mice,which are associated with the age of mice.

Rho-associated coiled coil protein kinase 1 involved in amyloid β-protein-induced damage in primary neurons of rats / 上海交通大学学报（医学版）

Objective • To investigate the role of Rho-associated coiled coil protein kinase 1 (ROCK1) in amyloid β-protein (Aβ) induced damage of rat hippocampal neurons. Methods • The rat primary neurons were treated with Aβ40 oligopeptides to establish a neurotoxicity model. Western blotting was used to detect the protein expression of ROCK1. Its activity was detected by the kit. Confocal laser scanning was used to observe the calcium signal in neurons, and apoptosis of neurons was detected by TUNEL assay. Y-27632, an inhibitor of ROCK1, was added into the culture medium in order to observe its effect on Aβ40. Results • Aβ40 (10 μmol/L) could significantly induce calcium overload, increase ROCK1 expression and activity, and promote apoptosis in primary neurons. Furthermore, ROCK1 inhibitor could decrease all the effect induced by Aβ40. Conclusion • ROCK1 is involved in both Aβ- induced neuronal calcium overload and neurotoxicity, and ROCK1 inhibitor can antagonize the toxic effects of Aβ.

Clearance effect of IL-33 on amyloid β-protein in brain tissue of model rats with Alzheimer's disease and its mechanism / 吉林大学学报(医学版)

Objective: To explore the influence of interleukin-33 (IL-33) on the abilities of spatial learning and memory and the levels of amyloid β-protein (Aβ) and helper T cell 2 (Th2) in the model rats with Alzheimer's disease (AD), and to clarify the effect of IL-33 in the scavenging of Aβ in brain tissue of the model rats with AD and its mechanism. Methods; A total of 70 SD male rats were randomly divided into normal control group (intragastrical administration of double steaming water and subcutaneous injection of D-galactose), model group, sham operation group and extremely low dose, low dose, middle dose, high dose (0.01, 0.10, 1.00, and 10. 00 mg · L-1) of IL-33 groups (intragastrical administration of AlCl3 and subcutaneous injection of D-galactose); 10 rats in each group. The agents were administered once daily for 60 d. After the last administration, step down test and Morris water maze experiment were used to detect the latency, the mistake times, the escape latencies, the resident time in target quadrant, and the swimming distance. After the behavioral testing, IL-33 was injected intracerebroventricularly for each dose of IL-33 group, and saline was injected intracerebroventricularly in the rats in sham operation group. ELISA method was used to detect the levels of Aβ and Th2 in brain tissue of the rats. Results: The results of step down test showed that the latency of the rats in model group was obviously shortened (t=8. 154, P<0. 01) and the mistake times were significantly increased compared with normal control group (t= 4. 579, P<0. 01). The results of Morris water maze test showed that the escape latency was obviously prolonged (t= 27. 810, P <0. 01), the resident time in target quadrant and the distance of swimming of the rats in model group were significantly reduced compared with normal control group (t= 3. 767, P<0. 01; t= 1. 973, P< 0. 05). The ELISA results showed that the level of Aβ in brain tissue of the rats in model group was significantly increased (t= 3. 222, P<0. 05) compared with normal control group, and the levels of Aβ in the brain tissue of the rats in low, middle and high doses of IL-33 groups were significantly reduced compared with model group (P< 0. 05). The level of Th2 in brain tissue of the rats in model group was significantly decreased compared with normal control group (t=4. 646, P<0. 01), and the levels of Th2 in brain tissue of the rats in low, middle and high doses of IL-33 groups were significantly increased compared with model group (P<0. 05). Conclusion: IL-33 has a scavenging effect on Aβ in brain tissue of the AD model rats, and its mechanism may be related to the increase of Th2 level in brain tissue of the rats.

Protective effect of total alkaloids of Corydalis Ochotensis on rats with Alzheimer's disease induced by β-amyloid protein / 吉林大学学报(医学版)

Objective:To explore the influence of total alkaloids of Corydalis Ochotensis(TAOCO) on the behavior and pathomorphology of brain tissue of the rats with Alzheimer's disease(AD) induced by β-amyloid protein 25-35(Aβ25-35),and to clarify its therapeutic effects on the AD rats.Methods:The Wistar rats were divided into mormal control group(treated with 0.5 mL·100 g-1 distilled water) (n=9),model group(treated with 0.5 mL·100 g-1 distilled water)(n=9),positive drug group(treated with 1.75 mg·kg-1 donepezil hydrochloride)(n=9),and low,middle and high doses (treated with 2.0,4.0 and 8.0 mg·kg-1) of TAOCO groups(n=8,n=9,n=9).The rat AD models were made by injecting Aβ25-35 into hippocampus.On the 14th day after operation,the rats were administered for 7 d.Morris water maze test was used to detect the spatial learning and memory ability of the rats;dark avoidance task was used to observe the passive avoidance ability of the rats;the pathomorphology of the cerebral cortex and hippocampus of the rats were detected.Results:The Morris water maze test results showed that compared with model group,the latency to platform of the rats in low dose of TAOCO group was decreased on the 4th and 5th days(P0.05).Compared with model group,there was no obvious improvement of the cerebral cortex and hippocampus injury of the rats in low and middle doses of TAOCO groups.In high dose of TAOCO group,the cerebral cortex and hippocampus injury of the rats were significantly improved.Conclusion:TAOCO can improve the learning and memory function of the AD rats and reduce the pathological injury of brain tissue of AD rats.

Effects of Mitochondrial Unfolded-Protein Response on Aggregation Toxicity of Amyloid-β Protein / 中山大学学报(医学科学版)

[Objective] To investigate the effects of mitochondrial unfolded-protein response (UPRmt) on the aggregation toxicity of Aβ protein in Alzheimer's disease (AD).[Methods] By cloning the mitochondrial outer membrane tomm-22,inner membrane E04A4.5 and atfs-1 genes of Caenorhabditis elegans (C.elegans) and constructing the L4440 interference vectors,HT115 competent cells were transformed to prepare tomm-22,E04A4.5 and atfs-1 RNAi bacteria.The effects of tomm-22 and E04A4.5 RNAi on the process of paralysis were investigated through transgenic AD disease models CL4176 and CL2006.The life span of wild type N2 C.elegans was observed after RNAi of tomm-22 and E04A4.5.The regulatory role of ATFS-1 signaling by atfs-1 RNAi in inhibition of Aβ protein aggregation was detected.The dynamic changes of UPRmt in transgenic SJ4100 nematode and the autophagy level in transgenic DA2123 nematodes were analyzed by tomm-22 and E04A4.5 RNAi.[Results] We successfully established the UPRmt model by cloning mitochondrial tomm-22 and E04A4.5 of C.elegans and further constructing RNAi bacteria,and showed that they can suppress aggregation toxicity of Amyloid-β (Aβ) protein in AD model CL4176,and slow down paralysis process.The life span of wild type N2 was significantly shortened after feeding with the tomm-22 and E04A4.5 RNAi bacteria.At the same time,the progressive paralysis AD model CL2006 shows a delayed paralysis in the early stage of life cycle but get acceleration in the late.These results illustrate that the UPRmt can alleviate the mitochondrial stress and improve the function of mitochondria at least in the short term.The atfs-1 RNAi confirmed that delayed paralysis process of AD model CL4176 is not directly related to the ATFS-1 signal.However,tomm-22 and E04A4.5 RNAi can gradually increase the UPRmt response and induce the expression level of autophagy-related molecules LGG-1,suggesting that tomm-22 and E04A4.5 RNAi may play a role in delaying the AD disease process by enhancing the activity of autophagy in C.elegans.[Conclusions] The study found that the UPRmt can inhibit the accumulation of A β protein by coordinating the signal transduction between mitochondria and nucleus,and can help to restore mitochondria and even intracellular protein homeostasis for protecting the normal physiological function of cells,and also provides new targets for prevention and treatment of neurodegenerative diseases such as AD.

Effect of Early Moxibustion Intervention on the Expression of Amyloid β-protein1-40 in 5XFAD Transgenic Mice / 上海针灸杂志

Objective To explore effect of early moxibustion intervention on cerebral Aβ1-40 in a mouse model of Alzheimer disease (AD) and the mechanism of action of moxibusion in preventing and treating AD.Method Gene phenotype in transgenic AD passage mice was identified using PCR. One and a half-month-old female Tg6799 transgenic mice were randomly allocated, including nine mice to a model group and eight mice to a treatment group. Nine C57BL/6J wild type female mice of the same age and background constituted a normal control group. Wheat-grain-sized moxa cone moxibustion on bilateral points Xinshu(BL15) and Shenshu(BL23) was given to the treatment group. After the completion of treatment, Aβ1-40 expression in mouse frontal cortex and hippocampal region was determined using the immunohistochemical method.Result Aβ1-40 expression in mouse frontal cortex and hippocampal region decreased significantly in the treatment group compared with the model group (P<0.01,P<0.05). Conclusion Early moxibustion intervention can decrease cerebral Aβ1-40 expression and delay AD pathological process in a mouse model of AD.

Effect of Early Moxibustion Intervention on the Expression of Amyloid β-protein1-40 in 5XFAD Transgenic Mice / 上海针灸杂志

Objective To explore effect of early moxibustion intervention on cerebral Aβ1-40 in a mouse model of Alzheimer disease (AD) and the mechanism of action of moxibusion in preventing and treating AD.Method Gene phenotype in transgenic AD passage mice was identified using PCR. One and a half-month-old female Tg6799 transgenic mice were randomly allocated, including nine mice to a model group and eight mice to a treatment group. Nine C57BL/6J wild type female mice of the same age and background constituted a normal control group. Wheat-grain-sized moxa cone moxibustion on bilateral points Xinshu(BL15) and Shenshu(BL23) was given to the treatment group. After the completion of treatment, Aβ1-40 expression in mouse frontal cortex and hippocampal region was determined using the immunohistochemical method.Result Aβ1-40 expression in mouse frontal cortex and hippocampal region decreased significantly in the treatment group compared with the model group (P<0.01,P<0.05). Conclusion Early moxibustion intervention can decrease cerebral Aβ1-40 expression and delay AD pathological process in a mouse model of AD.