益肾健脾通络方调节Th17/Treg失衡改善DKD小鼠肾损伤的作用机制研究  

Study on the Mechanism of Yishen Jianpi Tongluo Prescription Regulating Th17/Treg Imbalance to Improve Kidney Injury in DKD Mice

作　　者：俞烨晨 王旭[2] 杨帆 邢科 任丽红 徐广飞[1] YU Ye-chen;WANG Xu;YANG Fan;XING Ke;REN Li-hong;XU Guang-fei(Nantong Affiliated Hospital of Nanjing University of Chinese Medicine,Nantong 226001,China;The First Clinical College of Nanjing University of Chinese Medicine,Nanjing 210029,China;General Hospital of PLA Eastern Theater Command,Nanjing 210002,China)

机构地区：[1]南京中医药大学南通附属医院,江苏南通226001 [2]南京中医药大学第一临床医学院,江苏南京210029 [3]中国人民解放军东部战区总医院,江苏南京210002

出　　处：《中药材》2024年第3期713-718,共6页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金项目(81973796);南通市科技局基础科学研究计划-青年科技人才创新专项(JC12022084);南通市中医联盟课题-重点项目(TZYK202203)。

摘　　要：目的:探讨益肾健脾通络方对糖尿病肾脏病(DKD)小鼠辅助性T细胞17(Th17)/调节性T细胞(Treg)平衡及Notch信号通路的影响。方法:SPF级雄性C57BL/6N小鼠60只,随机选取10只为正常对照组,剩余50只小鼠腹腔注射链脲佐菌素构建DKD小鼠模型。将造模成功的小鼠随机分为5组,分别为模型组、厄贝沙坦(25 mg/kg)组及益肾健脾通络方高(生药5.2 g/kg)、中(生药2.6 g/kg)、低(生药1.3 g/kg)剂量组。各组小鼠均每天固定时间灌胃给药,连续12 w,正常对照组和模型组灌胃相应体积生理盐水。生化分析仪检测各组小鼠FBG、HbA1c、TC、LDL、Scr、BUN及尿液中mALB、UTP水平;通过HE、Masson、PAS染色观察小鼠肾组织病理学变化;ELISA检测血清NGAL、Lp-PLA2水平;流式细胞术检测外周血单个核细胞中Th17、Treg细胞的比例;Western Blot检测肾组织中Notch信号通路的Notch1、Jagged1蛋白表达。结果:给药12 w后,与正常对照组比较,模型组小鼠肾组织系膜细胞空泡化,大量肾小管管腔肿胀破裂,基底膜增厚;FBG、HbAIc、TC、LDL、Scr、BUN、NGAL、Lp-PLA2和尿液中mALB、UTP水平及肾组织中Notch1、Jagged1蛋白表达显著升高,Treg比例显著降低,Th17比例显著升高(P<0.05)。与模型组比较,厄贝沙坦组和益肾健脾通络方高剂量组肾组织病理损伤明显改善;FBG、HbAIc、TC、LDL、Scr、BUN、NGAL、Lp-PLA2和尿液中mALB、UTP水平及肾组织中Notch1、Jagged1蛋白表达显著降低,Treg比例显著升高,Th17比例显著降低(P<0.05)。高剂量益肾健脾通络方的作用与厄贝沙坦相当。结论:益肾健脾通络方可能通过抑制Notch信号通路调节Th17/Treg失衡,减轻DKD小鼠肾损伤。Objective:To investigate the effects of Yishen jianpi tongluo prescription on the balance of helper T cell 17(Th17)/regulatory T cell(Treg)and Notch signaling pathway in diabetic kidney disease(DKD)mice.Methods:Sixty male C57BL/6N mice with SPF grade were selected,10 were randomly selected as normal control group,and the remaining 50 mice were injected with streptozotocin intraperitoneally to construct DKD model,then the mice were randomly divided into 5 groups,including model group,irbesartan(25 mg/kg)group and Yishen jianpi tongluo prescription high-dose(crude drug 5.2 g/kg),medium-dose(crude drug 2.6 g/kg)and lowdose(crude drug 1.3 g/kg)groups.The mice in each group were given the drug intragastric at a fixed time every day for 12 weeks,and normal control group and model group were given the corresponding volume of normal saline intragastric administration.The FBG,HbA1c,TC,LDL,Scr,BUN were detected by biochemical analyzer,and the levels of mALB and UTP in urine were determined.The renal histopathological changes were observed by HE,Masson and PAS staining.Serum NGAL and Lp-PLA2 levels were detected by ELISA.The proportion of Th17 and Treg in PBMC was measured by flow cytometry.The expressions of Notch1 and Jagged1 proteins in Notch signaling pathway in kidney tissue was detected by Western Blot.Results:After 12 weeks of administration,compared with normal control group,mesangial cells of kidney tissue in model group were vacuolated,a large number of renal tubules were swollen and ruptured,and basement membrane was thickened.Blood glucose,blood lipids,renal function,urinary microalbumin,urinary protein quantity and Notch1 and Jagged1 protein expressions in renal tissue were significantly increased,while Treg proportion was significantly decreased and Th17 proportion was significantly increased(P<0.05).Compared with model group,the pathological injury of kidney tissue in irbesartan group and Yishen jianpi tongluo prescription high-dose group was significantly improved,blood glucose,blood lipid,kidney function,ur

关 键 词：益肾健脾通络方 糖尿病肾脏病 TH17/TREG NOTCH信号通路 

分 类 号：R285.5[医药卫生—中药学]