虫草益肾方对UUO大鼠Notch1及相关microRNA表达影响

Effrct of Chongcao Yishen Decoction on Expression of Notch1 and Related Microrna in UUO Rats

目的:探讨虫草益肾方对单侧输尿管梗阻(Unilateral ureteral occlusion,UUO)模型大鼠Notch1及相关microRNA的调控作用。方法:40只SPF级雄性SD大鼠随机分为假手术组10只及造模组30只,造模组采用UUO法进行模型制备,术后随机分为模型组、虫草益肾方组(虫草组)与厄贝沙坦组,每组10只。分别给予蒸馏水、虫草益肾方水煎液及厄贝沙坦水溶液进行灌胃,连续灌胃14 d收集肾组织。Masson染色观察肾脏病理改变,采用Real Time PCR及Western Blot法分别检测Notch1 mRNA转录与蛋白表达。通过生物信息学分析以Notch1为靶点的microRNA并采用Real Time PCR法进行检测。结果:虫草益肾方可改善UUO大鼠肾脏的病理变化程度;模型组大鼠14 d肾组织中Notch1 mRNA及蛋白表达与假手术组相比明显增加(P<0.05),虫草组、厄贝沙坦组大鼠Notch1mRNA及蛋白表达与模型组相比均减少(P<0.05)。通过生物信息学分析,检索并筛选出以Notch1为靶基因的miR-34a、miR-29a进行Real Time PCR检测,结果显示与假手术组相比,模型组、虫草组及厄贝沙坦组miR-34a、miR-29a表达明显降低(P<0.05);与模型组相比,虫草组与厄贝沙坦组miR-34a、miR-29a表达升高,其中虫草组二者表达均明显升高(P<0.05)。结论:虫草益肾方可改善因输尿管梗阻所致的肾脏结构病理变化,其作用可能是通过上调纤维化肾组织中miR-34a、miR-29a表达而靶向抑制Notch通路来实现的。

Objective:To investigate the regulation of Chongcao Yishen Decoction on Notch1 and related microRNA in unilateral ureteral occlusion(UUO)model rats.Methods:40 SD rats were randomly divided into 10 sham operation groups and 30 model-building groups.The model group was prepared by UUO method,and randomly divided into model group,Chongcao Yishen Decoction group(Chongcao group)and Irbesartan group,with 10 rats in each group.The rats were given intragastric administration of distilled water,Chongcao Yishen Decoction and irbesartan aqueous solution,and kidney tissue were collected on the 14 th day.Masson staining was used to observe the pathological changes of the kidney.Notch1 mRNA transcription and protein expression were detected by Real Time PCR and Western Blot method.MicroRNAs targeting Notch1 were analyzed by bioinformatics and detected by Real Time PCR.Results:Chongcao Yishen Decoction can improve the degree of renal pathological changes in UUO rats.The expression of Notch1 mRNA and protein of the model group rats at 14 days was significantly increased compared with the sham operation group(P<0.05).The mRNA and protein expression of Notch1,in rats of Chongcao group and Irbesartan group at 14 days were decreased compared with the model group(P<0.05).Bioinformatics analysis was used to search and screen for miR-34 a and miR-29 a with Notch1 as target gene for Real Time PCR.The results showed that the expression of miR-34 a and miR-29 a was significantly decreased in the model group compared with the sham operation group.(P<0.05);Compared with the model group,the expression of miR-34 a and miR-29 a in the Chongcao group and the Irbesartan group was increased,and both of the expression in the Chongcao group was significantly increased(P<0.05).Conclusion:Chongcao Yishen Decoction can improve the pathological changes of kidney structure caused by ureteral obstruction,which may be achieved by up-regulating the expression of miR-34 a and miR-29 a in fibrotic kidney tissue and targeting the Notch pathway.