基于转录组测序技术探讨活血降糖饮干预糖尿病肾病的机制  

Mechanism of Huoxue Jiangtang Drink in the Intervention of Diabetic Nephropathy Based on Transcriptome Sequencing Technology

作　　者：刘雪梅[1] 刘德亮[1] 曾霖 李惠林[1] 赵恒侠[1] 张学文[3] LIU Xuemmei;LIU Deliang;ZENG Lin;LI Huilin;ZHAO Hengxia;ZHANG Xuewen(Shenzhen Traditional Chinese Medicine Hospital,Shenzhen 518033,China;The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine,Shenzhen 518033,China;Institute of National Master of Traditional Chinese Medicine,Shaanxi University of Chinese Medicine,Xianyang 712000,China)

机构地区：[1]广东省深圳市中医院,深圳5180331 [2]广州中医药大学第四临床医学院,深圳518033 [3]陕西中医药大学国医大师研究所,咸阳712000

出　　处：《世界中医药》2021年第14期2120-2128,共9页World Chinese Medicine

基　　金：国家自然科学基金资助项目(81774225)——活血降糖饮通过Scrib/YAP对胰岛β细胞脂性凋亡的影响及机制;深圳市科技计划项目(JCYJ20160428180924481);深圳市医疗卫生“三名工程”项目(SZSM201512043)。

摘　　要：目的:探讨活血降糖饮治疗糖尿病肾病的分子机制。方法:通过高脂饲料喂养加尾静脉注射链脉佐菌素(STZ)等建立糖尿病肾病大鼠模型,并将模型大鼠随机分为糖尿病肾病(DN)组、模型+双胍+中药组(DN+Met+ZY)、模型+双胍+卡托普利组(DN+Met+CP),给予相应药物进行灌胃治疗,并另设正常对照组,每组10只,干预16周后,观察各组大鼠空腹血糖(FBG)、血脂、血清尿素氮(BUN)、血肌酐(Scr)、24 h尿总蛋白(24 h UTP)及尿微量清蛋白(24 h mALB)的变化。观察肾组织病理改变,并对各组肾组织进行RNA-seq测序。结果:与模型组比较,中药组和卡托普利组可以显著下调糖尿病肾病大鼠FBG、TC、TG、LDL-C、Scr、BUN、24 h UTP、24 h mALB等水平,而HDL-C无明显变化;与卡托普利组之间比较,中药组FBG、TC、TG、LDL-C明显下降,HDL-c、Scr、BUN、24 h UTP、24 h mALB均无明显差别。与模型组比较,中药组和卡托普利组均可以显著改善糖尿病肾病大鼠的肾脏病理学变化。转录组测序结果显示模型组对比正常组与中药组对比模型组转录本中基因总量接近,其中表达差异最明显的双向调控基因共筛选出10个;京都基因与基因组百科全书(KEGG)前20条显著富集通路分析显示模型组对比正常组与中药组对比模型组共同作用的信号通路有代谢相关通路及cAMP信号通路。结论:活血降糖饮联合二甲双胍能改善糖尿病肾病大鼠的糖脂代谢紊乱,减少尿白蛋白的排泄,改善肾功能和肾脏病理改变,延缓糖尿病肾病病情进展,其作用机制可能是通过Cftr、Pdk4、Angptl4、Hmgcs2等多靶点介导多通路实现的。Objective:To explore the molecular mechanism of Huoxue Jiangtang Decoction in the treatment of diabetic nephropathy.Methods:The diabetic nephropathy model was established by feeding high fat diet and injecting streptozotocin(STZ)through tail vein.The model rats were randomly divided into a diabetic nephropathy(DN)group,a model+biguanide+traditional Chinese medicine(DN+Met+ZY)model+biguanide+captopril(DN+Met+CP)group.The corresponding drugs were given by gavage,and another normal control group was set up,with 10 rats in each group.The changes of fasting blood glucose(FBG),blood lipid,serum urea nitrogen(BUN),serum creatinine(SCR),24 h urinary protein(24 h UTP)and urinary microalbumin(24h mAlb)were observed.The pathological changes of renal tissue were observed,and RNA-seq sequencing was performed in each group.Results:Compared with the model group,the levels of FBG,TC,TG,LDL-C,SCR,bun,24 h UTP and 24 h mAlb were significantly decreased in the Chinese medicine group and captopril group,but HDL-C had no significant change;compared with captopril group,FBG,TC,TG and LDL-C in Chinese medicine group decreased significantly,while there were no significant difference in HDL-C,SCR,bun,24 h UTP and 24 h mAlb.Compared with the model group,the Chinese medicine group and captopril group can significantly improve the renal pathological changes of diabetic nephropathy rats.The results of transcriptome sequencing showed that the total number of genes in the transcripts of model group vs normal group and traditional Chinese medicine group vs model group was close,and 10 bidirectional regulatory genes with the most obvious expression difference were screened out;the analysis of the first 20 KEGG rich pathways showed that the metabolic pathway and cAMP signaling pathway were involved in the model group vs the normal group and the Chinese medicine group vs the model group.Conclusion:Huoxue Jiangtang Drink combined with metformin can improve the disorder of glucose and lipid metabolism,reduce the excretion of urinary albumin,improve

关 键 词：活血降糖饮 糖尿病肾病 转录组测序技术 二甲双胍 糖尿病 链脲佐菌素 卡托普利 

分 类 号：R255.4[医药卫生—中医内科学] R285.5[医药卫生—中医学]