脾虚模型大鼠肠道菌群分布及时效性研究  

Study on the Distribution and Timeliness of Intestinal Flora in Spleen Deficiency Model Rats

作　　者：郑昊龙 陈丝 宋囡[1,3] 曹慧敏[1,3] 吴瑶 王杰[1] 贾连群[1,3] ZHENG Haolong;CHEN Si;SONG Nan;CAO Huimin;WU Yao;WANG Jie;JIA Lianqun(Ministry of Education Key Laboratory of Theory and Application of Visceral Manifestations of Traditional Chinese Medicine;General Hospital of North Theater Command;Center of Traditional Chinese Medicine Innovation Engineering Technology,Liaoning University of Traditional Chinese Medicine)

机构地区：[1]辽宁中医药大学中医脏象理论及应用教育部重点实验室,辽宁省沈阳市110847 [2]北部战区总医院 [3]辽宁中医药大学中医药创新工程技术中心

出　　处：《中医杂志》2020年第14期1262-1267,共6页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81774022);辽宁省自然基金指导计划(2019-ZD-0966)。

摘　　要：目的应用肠道微生物16S rDNA测序技术探讨脾虚对肠道微生物的影响及时效性。方法将48只大鼠随机分为对照组和脾虚组,每组又分为2、10、16周3个时间点,每组每个时间点8只大鼠,雌雄各半。脾虚组采用饮食不节加力竭游泳法建立脾虚模型。分别于0、2、10、16周时检测雌雄大鼠体重,于2、10、16周时检测大鼠尿D-木糖排泄率、血清淀粉酶活力。采用肠道菌群Beta多样性分析、Alpha多样性分析比较对照组和脾虚组不同时间点肠道微生物丰度和物种组成变化情况。结果与对照组同时间相比,脾虚组2、10、16周时雌、雄性大鼠体重均显著减轻(P<0.05或P<0.01);脾虚组2、10、16周时大鼠尿D-木糖排泄率显著降低,2、10周时血清淀粉酶亦降低(P<0.05或P<0.01)。16S rDNA测序结果显示,肠道菌群Beta多样性分析中线性判别分析(LDA)>2且在该组中丰度显著高的差异物种对照组有8种,脾虚组2周时有14种,10周时有27种,16周有8种。Alpha多样性分析结果:与对照组相比,脾虚组2、10周时大鼠肠道微生物ACE、Observed指数呈上升趋势,16周时有下降趋势,但差异无统计学意义(P>0.05)。脾虚组16周时大鼠ACE、Observed指数显著低于本组2、10周(P<0.05)。与对照组相比,脾虚组2周时大鼠肠道菌群α-变形菌纲水平显著升高(P<0.01)。与脾虚组2周时相比,脾虚组10、16周时α-变形菌纲和放线菌纲水平下降(P<0.05或P<0.01),各时间点疣微菌纲差异无统计学意义(P>0.05)。结论脾虚条件下大鼠体重减轻,代谢功能降低,其发生机制可能与肠道菌群丰度及物种数量变化相关,且脾虚状态可随时间变化逐渐减轻。Objective To investigate the effects and timeliness of spleen deficiency on intestinal microorganisms using 16 S rDNA sequencing technology of intestinal microorganisms.Methods A total of 48 rats were randomly divided into a control group and a spleen-deficiency group,and each group was divided into 3 subgroups of time points 2,10,and 16 weeks.Each group had 8 rats at each time point,half male and half female.The spleen-deficiency group established a model of spleen-deficiency by irregular diet and strenuous swimming.Body weights of male and female rats were measured at 0,2,10,and 16 weeks,and urine D-xylose excretion rate and serum amylase activity were detected at 2,10,and 16 weeks.Intestinal microbiota Beta diversity analysis and Alpha diversity analysis were used to compare changes in intestinal microbial abundance and species composition at different time points in the control group and the spleen deficiency group.Results Compared with the control group at the same time,the weight of female and male rats in the spleen-deficiency group was significantly reduced at 2,10,and 16 weeks(P<0.05 or P<0.01);the urine D-xylose excretion rate of rats in the spleen-deficiency group was decreased significantly at 2,10,and 16 weeks,and serum amylase also decreased at 2 and 10 weeks(P<0.05 or P<0.01).16 S rDNA sequencing results showed that in analysis of intestinal flora Beta diversity,there were 8 species in the control group with linear discriminant analysis(LDA)>2 and with significantly higher abundance in the group.The number of species in the spleen deficiency group was 14 at 2 weeks,27 at 10 weeks and 8 at 16 weeks.Alpha diversity analysis results showed that compared with the control group,the intestinal microbial ACE and Observed index of rats in the spleen-deficiency group showed an upward trend at 2 and 10 weeks,and a downward trend at 16 weeks,but there was no statistically significant difference(P>0.05).The ACE and Observed index of rats in the spleen-deficiency group at 16 weeks were significantly lower than

关 键 词：脾虚 肠道菌群 16S rDNA测序 时效性 

分 类 号：R-332[医药卫生]