加减薯蓣丸对阿尔茨海默病小鼠模型肠道菌群和HMGB1通路相关蛋白的影响

Effects of Modified Shuyuwan(薯蓣丸) on Gut Microbiota and HMGB1 Pathway-Related Proteins in Alzheimer’s Mice

目的:探讨加减薯蓣丸通过高迁移率族蛋白B1(HMGB1)通路相关蛋白和肠道菌群对阿尔茨海默病小鼠的影响。方法:将40只APP/PS1小鼠分成模型对照组、加减薯蓣丸14、56 g/kg组及盐酸多奈哌齐0.001 g/kg组,每组10只,另将10只同月龄野生型C57BL/6J小鼠作为正常对照组。小鼠灌胃给予相应药物或生理盐水,1次/d,连续4 w。药物干预结束后通过新物体识别实验测定小鼠学习记忆能力,尼氏染色观察小鼠海马组织神经元变化,菌落计数法检测小鼠肠道内容物中乳酸杆菌和双歧杆菌数量变化,ELISA法检测小鼠脑组织中的白细胞介素-1β (IL-1β)和肿瘤坏死因子-α (TNF-α)含量,Western blot法检测小鼠海马组织中HMGB1、Toll样受体4(TLR4)和核因子κB p65(NF-κB p65)蛋白的表达。结果:与正常对照组比较,模型对照组小鼠新物体偏爱指数、海马组织尼氏体数量、肠道乳酸杆菌和双歧杆菌数量均明显降低,脑组织中IL-1β和TNF-α含量明显升高,海马组织中HMGB1、TLR4和NF-κB p65蛋白表达明显上调(P<0.05);与模型对照组比较,加减薯蓣丸56 g/kg组小鼠新物体偏爱指数、海马组织尼氏体数量、肠道乳酸杆菌和双歧杆菌数量均明显升高,脑组织中IL-1β和TNF-α含量明显降低,海马组织中HMGB1、TLR4和NF-κB p65蛋白表达明显下调(P<0.05)。结论:加减薯蓣丸能改善阿尔茨海默病小鼠的学习记忆能力,其具体机制可能和调控肠道菌群数量及HMGB1通路相关蛋白有关。

Objective:To investigate the effects of modified Shuyuwan(薯蓣丸)on the high mobility group protein B1(HMGB1)pathway-related proteins and gut microbiota of Alzheimer's mice.Methods:Forty APP/PS1 mice were assigned into a model group,modified Shuyuwan groups(14 g/kg and 56 g/kg),and a donepezil hydrochloride(0.001 g/kg)group,with 10 mice in each group.Another 10 wild-type C57BL/6J mice of the same age were selected as the normal control group.The mice in each group were administrated with corresponding drugs or normal saline by gavage once a day for 4 consecutive weeks.After the drug intervention,the learning and memory ability of mice was examined by novel object recognition task.Nissl staining was used for observation of the changes of neurons in the hippocampus of mice.Colony counting method was used to count Lactobacillus and Bifidobacterium in the intestinal contents of mice.Enzyme-linked immunosorbent assay(ELISA)was employed to determine the levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the brain tissue of mice.Western blotting was employed to determine the protein levels of HMGB1,toll-like receptor 4(TLR4),and nuclear factor-kappa B p65(NF-κB p65)in mouse hippocampus.Results:Compared with the normal control group,the model group showed decreased novel object preference index,number of Nissl bodies in the hippocampus,and number of Lactobacillus and Bifidobacterium(P<0.05),elevated levels of IL-1βand TNF-αin the brain tissue(P<0.05),and up-regulated protein levels of HMGB1,TLR4,and NF-κB p65 in the hippocampus(P<0.05).Compared with the model group,modified Shuyuwan(56 g/kg)increased the novel object preference index,number of Nissl bodies in the hippocampus,and number of Lactobacillus and Bifidobacterium(P<0.05).In addition,it lowered the levels of IL-1βand TNF-αin the brain tissue(P<0.05)and down-regulated the protein levels of HMGB1,TLR4,and NF-κB p65 in the hippocampus(P<0.05).Conclusion:Modified Shuyuwan may improve the learning and memory ability of Alzheimer's mice by regulating the number of gut microbiota and the HMGB1 pathway-related proteins.