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AIM:To investigate the effect of high mobility group box-1 protein (HMGB1) on the expression of nuclear factor-κB ( NF-κB) in BV-2 cells stimulated with amyloid β-protein ( Aβ) 25-35 .METHODS:Cultured BV-2 cells in logarithmic growth phase were divided into 4 groups:normal cell group ( without any treatment ) , model group ( treated with Aβ25-35 at 40 μmol/L) , RNA interference ( RNAi) group ( conducted with HMGB1-siRNA followed by Aβ25-35 stimula-tion) and solvent control group (treated with 0.1% DMSO).After treatment with Aβ25-35 for 24 h, the protein levels of HMGB1 and NF-κB in BV-2 cells were determined by Western blot .RESULTS:Aβ25-35 at 40μmol/L was used to stimu-late BV-2 cells.The GFP fluorescence-tagged HMGB1-siRNA (30 nmol/L) was used to transfect BV-2 cells and its trans-fection efficiency was about 80%~90%.The results of Western blot showed that the protein level of HMGB 1 was signifi-cantly decreased after the interference of siRNA fragment (P<0.05).The protein levels of HMGB1 and nucleic NF-κB p65 were dramatically increased in BV-2 cells stimulated with Aβ25-35(P<0.05).After RNA interference with HMGB1, the expression of HMGB1 and nucleic NF-κB p65 were significantly decreased in BV-2 cells stimulated with Aβ25-35 ( P<0. 05).CONCLUSION:RNA interference with HMGB1 reduces the expression of nucleic NF-κB in BV-2 cells stimulated with Aβ25-35 .
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OBJECTIVE:To study the effects of aripiprazole on PC12 cell injury induced by amyloid β-protein(Aβ25-35)and its mechanism. METHODS:PC12 cells were randomized into normal control group,model group (20 μmol/L Aβ25-35),aripiprazole low-concentration,medium-concentration and high-concentration groups(5,10,20 μmol/L aripiprazole+20 μmol/L Aβ25-35). These groups were cultured with culture medium containing relevant medicine for 48 h,with 6 wells in each group. The viability(optical density value)of PC12 cell was measured by MTT assay,and PC12 cell apoptosis was measured by Hoechst staining. The activi-ties of Caspase-3 and Caspase-9 were determined by spectrophotometry. The protein expression of Bcl-2,Bax and PI3K and the phosphorylation of Akt were assayed by Western blot assay. RESULTS:Compared with normal control group,optical density value of model group was decreased while apoptotic rate was increased;the activities of Caspase-3 and Caspase-9,and the protein expres-sion of Bax were increased;the protein expression of Bcl-2 and PI3K,the phosphorylation of Akt were decreased(P<0.01). Com-pared with model group,optical density value of aripiprazole low-concentration,medium-concentration and high-concentration groups were increased,while apoptotic rate and the activities of Caspase-3 and Caspase-9 were decreased;the protein expression of Bcl-2 and PI3K and the phosphorylation of Akt were enhanced;while the protein expression of Bax were decreased in aripiprazole medium-concentration and high-concentration groups(P<0.05 or P<0.01). CONCLUSIONS:Aripiprazole can suppress cell apop-tosis of PC12 cell induced by Aβ25-35,which is related to activating PI3K/Akt signal pathway.
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Objective To observe the effect of moxibustion on the learning ability and memory of rat models of Alzheimer’s disease (AD) and the expression of amyloidβ protein (Aβ)-42, interleukin (IL)-1β, and IL-2, for unveiling the function and mechanism of moxibustion in treating AD.Method Forty-eight healthy male Sprague-Dawley rats were randomized into a sham-operation group, a model group, a Western medication group, and a moxibustion group, 12 rats in each group. The AD model rats were established by injection of Streptozotocin (STZ) into bilateral ventricles. From the 10th day after the operation, the moxibustion group started to receive moxibustion at Baihui (GV 20), Mingmen (GV 4), Changqiang (GV 1), and Guanyuan (CV 4); meanwhile, the Western medication group received Donepezil hydrochloride via intragastric administration. After 30-day treatment, the learning and memory ability was tested by using water maze, the hippocampal Aβ-42 was examined by immunohistochemical method, and the expression of hippocampal IL-1β and IL-2 was by double-antibody sandwich ELISA.Result Compared to the model group, moxibustion significantly down-regulated the levels of Aβ-42 (P<0.05) and IL-1β protein (P<0.05), up-regulated the level of IL-2 protein (P<0.05) in hippocampus, and markedly improved the learning and memory of AD rats (P<0.05).Conclusion Moxibustion can enhance the immunity and learning ability, which is plausibly related to the down-regulation of IL-1β, up-regulation of IL-2, and improvement of Aβ-42 in hippocampus.
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[ ABSTRACT] AIM:To observe the treatment effect and its immune regulation of human amnion epithelial cells ( hAECs) on Alzheimer’ s disease ( AD)-like pathology rat model.METHODS: The hAECs were isolated from amnion with trypsin digestion, and the phenotype of hAECs was analyzed by flow cytometry.SD rats ( n=48) were randomly divid-ed into sham control group, model group, medium group and hAECs group.AD-like pathology rat model was induced by bilateral intraventricular injection of lipopolysaccharide (LPS).hAECs (5 ×105) were injected into the hippocampus of the AD-like pathology rats.At 2 weeks after transplantation, the animals were tested by Morris water maze to observe the function of learning and memory.The pathological change of the brain was observed by HE staining.The expression of am-yloid β-protein 42 (Aβ42) and Tau protein and the level of acetylcholine (ACh) in the injury brain were determined by immunohistochemistry.The survival and differentiation of hAECs in the hippocampus were measured by immunofluorescent technique.The percentages of lymphocyte subsets in the peripheral blood mononuclear cells were analyzed by flow cytome-try.The contents of serum cytokines were detected by cytometric bead array.RESULTS:Compared with model group and medium group, hAECs group showed shortened escape latency ( P<0.01) , increased frequency of going through the plat-form (P<0.05), reduced loss of hippocampal neurons, decreased expression of Tau protein and Aβ42 in the hippocampus (P<0.05), increased ACh level in the hippocampus (P<0.05), decreased percentages of Th1 and Th17 subsets, in-creased percentages of Th2 and Treg cells ( P<0.05) , decreased concentrations of IFN-γand IL-2 in the serum, and in-creased concentration of IL-4 ( P<0.05 ) .CONCLUSION: hAECs improve the cognitive learning and memory function and alleviate pathologic damage of hippocampus through immune regulation in AD-like pathology rats.
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Objective To observe the influence of XueShuanXinMaiNing(XSXMN)in the behavior and structures of cerebral cortex and hippocampus of the rats withβamyloid protein(Aβ)-induced Alzheimer’s disease(AD),and to explore its therapeutic effects on the rat AD.Methods 100 male Wistar rats were selected.According to weight, the rats were randomly divided into sham operation group, model group, positive drug group (donepezil hydrochloride,1.75 mg· kg-1 ),XSXMN 1.1 g· kg-1 group and XSXMN 2.2 g· kg-1 group. The rat AD models were made by injecting Aβinto hippocampus.After oral administration for 15 d,Morris water maze test, dark avoidance task and pathology test were performed.Results In Morris water maze test,compared with model group,the latency and swimming distance to platform of the rats in XSXMN 1.1 g·kg-1 group were decreased on the 2nd,4th and 5th day(P<0.05 or P<0.01);in XSXMN 2.2 g·kg-1 group,the latency to platform of the rats were decreased from the 3nd to 6th day(P<0.05 or P<0.01),the swimming distances to platform of the rats were decreased from the 3rd to 5th day(P<0.05 or P<0.01).On the 7th day,in XSXMN groups,the times of passing platform,time of staying on platform,distance of staying on platform,time of staying in effective area, distance of staying in effective area, time of staying on platform/total time, distance of staying on platform/total distance,time of staying on platform/total time were all increased significantly(P<0.05 or P<0.01)within 90 s. In dark avoidance task,compared with model group,the error latency and the error times of the rats in XSXMN groups had no obvious change on the 2nd day.The pathological results showed that there were degeneration nerve cells and necrosis nerve cells in the rat cerebral cortex in XSXMN groups,while in the rat hippocampus there were less number of nerve cells with obscure cell layer and many degeneration and necrosis cells were found;compared with model group,there was no obvious improvement.Conclusion XSXMN can improve the learning and memory function of the AD rats.