参芪复方对糖尿病血糖波动模型大鼠胰腺组织生物钟相关基因与肠道菌群的影响

Effects of Shenqi Compound (参芪复方) on Biological Clock Related Genes and Intestinal Flora in Pancreatic Tissue of Diabetic Glucose Fluctuation Model Rats

目的探讨参芪复方对糖尿病血糖波动模型大鼠血糖稳态的影响及可能作用机制。方法30只自发性2型糖尿病GK大鼠随机分为模型组、参芪复方组和西格列汀组,每组10只,均予高脂饲料喂养4周建立血糖波动模型;10只Wistar大鼠设为正常组,予普通饲料喂养4周。造模成功后参芪复方组给予参芪复方浸膏1.44g/(kg·d)灌胃,西格列汀组给予西格列汀片悬液16mg/(kg·d)灌胃,正常组、模型组给予生理盐水5ml/(kg·d)灌胃,均连续8周。大鼠尾静脉取血,测1日内8:00、10:00、14:00、16:00、18:005次血糖水平,计算一日血糖平均水平(MBG)、最大血糖波动幅度(LAGE)、平均血糖的标准差(SDBG);实时荧光定量PCR法检测胰腺组织生物钟相关基因Id 2、Usp 2、CRY1、DBP mRNA表达;运用16S rDNA基因测序分析各组大鼠肠道菌群属水平分布情况。结果与正常组比较,其余各组大鼠MBG、SDBG升高,模型组和参芪复方组LAGE亦升高(P<0.01);与模型组比较,参芪复方组和西格列汀组大鼠MBG、SDBG、LAGE均降低(P<0.01),西格列汀组在降低SDBG、LAGE方面优于参芪复方组(P<0.01)。与正常组比较,模型组大鼠Usp 2、CRY1 mRNA表达及丁酸菌属水平均降低(P<0.05或P<0.01);与模型组比较,参芪复方组和西格列汀组大鼠Id 2 mRNA表达降低,Usp2、CRY1 mRNA表达升高,拟杆菌属、丁酸菌属水平亦升高(P<0.05或P<0.01);各组DBP mRNA表达、布劳特氏菌属和罗氏菌属水平差异无统计学意义(P>0.05)。结论参芪复方可能通过调节血糖波动模型大鼠胰腺组织生物钟相关基因的表达及肠道菌群菌属的分布,从而调节血糖稳态。

Objective To investigate the effects of Shenqi Compound(参芪复方)on the blood glucose homeostasis of diabetic blood glucose fluctuation model rats and its possible mechanism.Methods A total of 40 spontaneous type 2 diabetic GK rats were randomly divided into a model group,a Shenqi Compound group and a sitagliptin group,with 10 rats in each group.All rats were fed with high-fat diet for 4 weeks to establish a blood glucose fluctuation model.Ten Wistar rats were set into the normal group and fed with ordinary feed for 4 weeks.After successful modeling,the Shenqi Compound group was administered with Shenqi Compound extract 1.44 g/(kg·d),and the sitagliptin group was administered with 16 mg/(kg·d)of sitagliptin tablet suspension.The normal group,and the model group were given intragastrically with 5 ml/(kg·d)saline for 8 consecutive weeks.Blood was taken from the rat tail vein,and blood glucose levels were measured 5 times during the day at 8:00,10:00,14:00,16:00,and 18:00,and the daily mean blood glucose(MBG)and largest amplitude of glycemic excursions(LAGE)were calculated,standard deviation of mean blood glucose(SDBG)were caculated;real-time fluorescent quantitative PCR was used to detect mRNA expression of biological clock-related genes Id 2,Usp 2,CRY 1,and DBP;16 S rDNA gene sequencing was used to analyze intestinal flora of each group of rats.Results Compared with the normal group,MBG and SDBG in the other groups were increased,and LAGE in the model group and the Shenqi Compound group was also increased(P<0.01).Compared with the model group,MBG,SDBG,and LAGE of the Shenqi Compound group and the sitagliptin group were all reduced(P<0.01).The sitagliptin group was superior to the Shenqi Compound group in reducing SDBG and LAGE(P<0.01).Compared with the normal group,the expression of Usp 2,CRY1 mRNA and butyric acid levels in the model group rats were reduced(P<0.05 or P<0.01).Compared with the model group,the rats in the Shenqi Compound group and the sitagliptin group had Id 2 mRNA expression decreased,Usp 2,CRY1 mRNA expression increased,and level of Bacteroides and Butyrica also increased(P<0.05 or P<0.01);There was no significant difference in the levels of DBP mRNA expression,Broutella,and Rhodium in all groups(P>0.05).Conclusion Shenqi Compound may regulate the blood glucose homeostasis by regulating the expression of biological clock-related genes and the distribution of intestinal flora in the pancreas tissue of rat models of blood glucose fluctuation.