基于脑-肠-微生物轴探讨益脾通腑法对帕金森病小鼠的干预机制  

Discussion on the intervention mechanism of Yipi Tongfu method on Parkinson’s disease mice based on brain-intestine-microorganism axis

作　　者：刘畅[1] 李常慧 王姝元 区益洲 缪晓路[2] LIU Chang;LI Changhui;WANG Shuyuan;OU Yizhou;LIAO Xiaolu(Guangzhou University of Chinese Medicine,Guangzhou 510006,Guangdong,China;Dongguan Hospital,Guangzhou University of Chinese Medicine,Dongguan 523005,Guangdong,China)

机构地区：[1]广州中医药大学,广东广州510006 [2]广州中医药大学东莞医院,广东东莞523005

出　　处：《现代中西医结合杂志》2022年第10期1320-1327,共8页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：广东省中医药局科研项目(20191347)。

摘　　要：目的基于脑-肠-微生物轴探讨益脾通腑法(术虎合剂)干预帕金森病小鼠的机制。方法将60只C57BL/6雄性小鼠随机分为正常组、模型组、白术组、虎杖组及术虎合剂组,每组12只。除正常组外,其余组每日给予1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)30 mg/kg腹腔注射进行帕金森病造模,连续7 d。造模成功后,白术组给予27.5 mg/mL白术汤剂(生药白术10 g)灌胃,虎杖组给予10 mg/mL虎杖汤剂(生药虎杖10 g)灌胃,术虎合剂组给予65 mg/mL术虎合剂(生药白术20 g和虎杖10 g)灌胃,空白组及模型组给予等量蒸馏水灌胃,均连续灌胃2周。末次灌胃后对各组小鼠进行行为能力检测,记录前爪步距、掉头时间、下杆时间;收集小鼠脑组织,采用HE和Nissl染色法进行病理观察;收集小鼠粪便,提取粪菌总DNA后进行16 s rRNA高通量测序并分析其结果。结果各组小鼠前爪步距比较差异均无统计学意义(P均>0.05);模型组小鼠掉头时间、下杆时间均明显长于正常组(P均<0.05),白术组、虎杖组、术虎合剂组小鼠掉头时间、下杆时间均明显短于模型组(P均<0.05),且术虎合剂组小鼠掉头时间明显短于虎杖组(P<0.05)。HE染色显示模型组小鼠神经元数目减少,各药物组小鼠神经元数量增加,其中术虎合剂组神经细胞数量最多;Nissl染色显示模型组小鼠黑质致密带神经元数目明显减少,黑质中尼氏小体的数量明显少于正常组(P<0.05);白术组和虎杖组小鼠神经细胞形态部分恢复,术虎合剂组小鼠神经细胞形态恢复正常,尼氏小体数量均明显多于模型组(P均<0.05)。粪菌测序:门分类水平,术虎合剂组厚壁菌门和变形菌门比例较模型组低,疣微菌门比例较模型组高;科分类水平,术虎合剂组毛螺菌科、疣微菌科比例较模型组高;属分类水平,术虎合剂组阿克曼菌属、Allobaculum比例较模型组高;术虎合剂组Shannon指数和Chao指数均明显高于模型组(P均<0.05),�Objective It is to explore the mechanism of Yipi Tongfu method(Zhuhu mixture)in the treatment of mice with Parkinson’s disease.Methods Sixty C57BL/6 male mice were randomly divided into normal group,model group,Atractylodes macrocephala Koidz group,Polygonum cuspidatum group,and Zhuhu mixture group(Atractylodes macrocephala Koidz and Polygonum cuspidatum mixture group),12 animals in each group.Except for the normal group,1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)was injected intraperitoneally at a dose of 30 mg/kg daily to establish models of Parkinson’s disease in the other groups,continuously treated for 7 days.After the models were successfully established,the Atractylodes macrocephala Koidz group was given 27.5 mg/mL Atractylodes macrocephala Koidz decoction(crude drug Atractylodes macrocephala Koidz 10 g)by gavage,the Polygonum cuspidatum group was given 10 mg/mL Polygonum cuspidatum decoction(crude drug Polygonum cuspidatum 10 g)by gavage,and the Zhuhu mixture group was given 65 mg/mL Zhuhu mixture(crude drug Atractylodes macrocephala Koidz 20 g and Polygonum cuspidatum 10 g)by gavage,the blank group and model group were given the same volume of distilled water by gavage,all the groups were treated for 2 consecutive weeks.After the last gavage,the behavioral ability of the mice in each group was tested,and the forepaw step distance,U-turn time and downswing time were recorded;the mouse brain tissue was collected for pathological observation by HE and Nissl staining methods;the mouse feces were collected to extract the total DNA of fecal bacteria for 16s rRNA high-throughput sequencing and the results were analyzed.Results There was no significant difference in the step distance of the front paw of the mice among each group(all P>0.05);the U-turn time and downswing time of the mice in the model group were significantly longer than those in the normal group(all P<0.05).The U-turn time and downswing time of the mice in the Atractylodes macrocephala Koidz group,Polygonum cuspidatum group and Zhuhu m

关 键 词：帕金森病 益脾通腑法 白术 虎杖 肠道菌群 

分 类 号：R-332[医药卫生]