柴胡桂枝干姜汤对2型糖尿病大鼠肠道菌群及短链脂肪酸的影响研究

Effects of Chaihu Guizhi Ganjiang decoction on gut microbiota and short-chain fatty acids in type 2 diabetes rats

目的:探讨柴胡桂枝干姜汤对2型糖尿病(T2DM)大鼠肠道菌群及短链脂肪酸的影响。方法:将Wistar大鼠随机分为正常组、模型组和柴胡桂枝干姜汤组。正常组8只,予正常饲料喂养,其余22只予高脂高糖饲料喂养6周后,腹腔注射链脲佐菌素(STZ)32 mg/kg进行T2DM造模,最后共有18只大鼠造模成功,将18只T2DM大鼠随机分为模型组和柴胡桂枝干姜汤组,每组9只。正常组和模型组予0.9%氯化钠溶液10 ml/kg进行灌胃,柴胡桂枝干姜汤组予柴胡桂枝干姜汤11.24 g/kg灌胃,每天给药2次,持续给药8周。使用罗氏血糖仪检测空腹血糖(FBG);ELISA法检测糖化血红蛋白(HbA1c);利用Novaseq 6000系统对肠道微生物16S rDNA的V3+V4区进行扩增测序;利用液相色谱-质谱检测粪便短链脂肪酸水平。结果:与正常组比较,模型组的FBG及干预8周后的HbA1c显著升高(P<0.05),干预前后的HbA1c差值显著增大(P<0.05),厚壁菌门、乳酸杆菌属的相对丰度显著下降(P<0.05),拟杆菌门、瘤胃球菌属的相对丰度显著增加(P<0.05),丁酸和异丁酸含量显著上调(P<0.05),而主要产丁酸菌如毛螺杆菌科、普雷沃氏菌科、双歧杆菌科和粪杆菌属的相对丰度无明显增加,差异无统计学意义(P>0.05),拟杆菌科的相对丰度出现下降(P<0.05)。与模型组比较,柴胡桂枝干姜汤组在干预6周后的FBG显著下降(P<0.05),干预前后的HbA1c差值显著减少(P<0.05),厚壁菌门、乳酸杆菌属的相对丰度显著增加(P<0.05),瘤胃球菌属的相对丰度显著下降(P<0.05),丁酸含量显著下降(P<0.05),毛螺杆菌科、普雷沃氏菌科、双歧杆菌科和拟杆菌科的相对丰度差异无统计学意义(P>0.05),粪杆菌属相对丰度显著上升(P<0.05)。结论:柴胡桂枝干姜汤能够改善2型糖尿病大鼠的FBG及HbA1c,其作用可能与增加乳酸杆菌属丰度、减少瘤胃球菌属丰度以及调节短链脂肪酸的代谢吸收有关。

Objective:To investigate the effects of Chaihu Guizhi Ganjiang decoction on gut microbiota and short-chain fatty acids in type 2 diabetes(T2DM)rats.Methods:Wistar rats were randomly divided into a normal group,a model group,and a Chaihu Guizhi Ganjiang decoction group.A normal group of 8 rats were fed with normal feed,while the remaining 22 rats were fed with high-fat and high sugar feed for 6 weeks.After intraperitoneal injection of Streptozotocin(STZ)at a dose of 32 mg/kg,T2DM models were established.Finally,a total of 18 rats were successfully established.The 18 T2DM rats were randomly divided into a model group and a Chaihu Guizhi Ganjiang decoction group,with 9 rats in each group.The normal group and model group were given 10 ml/kg of physiological saline by gavage,while the Chaihu Guizhi Ganjiang decoction group was given 11.24 g/kg of Chaihu Guizhi Ganjiang decoction by gavage twice a day for 8 weeks.Use a Roche blood glucose meter to measure fasting blood glucose(FBG);ELISA method for detecting glycated hemoglobin A1c(HbA1c);the Novaseq 6000 system to sequence the V3+V4 region of 16S rDNA from gut microbiota;liquid chromatography-mass spectrometry(LC-MS)to detect fecal short chain fatty acid levels.Results:Compared with the normal group,the FBG and HbA1c of the model group were significantly increased(P<0.05)after 8 weeks of intervention,and the difference in HbA1c before and after intervention was significantly increased(P<0.05).The relative abundance of Firmicutes and Lactobacillus genera were significantly decreased(P<0.05),while the relative abundance of Bacteroidetes and Ruminococcus genera were significantly increased(P<0.05).The content of butyric acid and isobutyric acid was significantly upregulated(P<0.05),while the main butyric acid producing bacteria such as Lachnospiraceae,Prevotellaceae,Bifidobacteriaceae,and Faecalibaculum did not show a significant increase,with no statistically significant difference(P>0.05),and even the relative abundance of Bacteroideaceae decreased(P<0.05).Compared with the model group,the Chaihu Guizhi Ganjiang decoction group showed a significant decrease in FBG after 6 weeks of intervention(P<0.05),a significant decrease in HbA1c difference before and after intervention(P<0.05),a significant increase in relative abundance of Firmicutes and Lactobacillus genera(P<0.05),a significant decrease in relative abundance of Ruminococcus genus(P<0.05),and a significant decrease in butyric acid content(P<0.05),while the main butyric acid producing bacteria such as Lachnospiraceae,Prevotellaceae,Bifidobacteriaceae and Bacteroideaceae did not significantly decrease,and the difference was not statistically significant(P>0.05),and the relative abundance of Faecalibaculum significantly increased(P<0.05).Conclusion:Chaihu Guizhi Ganjiang decoction can improve FBG and HbA1c in type 2 diabetes rats,which may be related to increasing the abundance of Lactobacillus,reducing the abundance of Ruminococcus and regulating the metabolism and absorption of short chain fatty acids.