中药复方益肾康调控Klotho/NF-κB信号通路防治糖尿病大鼠肾脏损伤的机制研究  

Study on the Mechanism of Chinese Herbal Compound Yishenkang(复方益肾康)Regulating Klotho/NF-κB Signaling Pathway to Prevent and Treat Renal Injury in Diabetes Rats

作　　者：张颖[1] 黄奡 宫成军[1] 张兰[1] ZHANG Ying;HUANG Ao;GONG Chengjun;ZHANG Lan(Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China)

机构地区：[1]辽宁中医药大学附属医院,辽宁沈阳110032

出　　处：《辽宁中医药大学学报》2023年第5期23-28,共6页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：全国名老中医药专家传承工作室建设项目(2022-304);辽宁中医附属医院“育苗工程”项目(YM202119)。

摘　　要：目的基于Klotho/NF-κB信号通路探讨中药复方益肾康保护肾脏的疗效机制。方法采用单次腹腔注射链脲佐菌素(53 mg/kg)构建糖尿病大鼠模型,将雄性SD大鼠随机分为正常对照组(10只)、模型组(16只)、厄贝沙坦组(15只)、益肾康组(15只)。干预4周后,收集尿液检测尿微量白蛋白/尿肌酐(UACR)。干预8周后,测量大鼠体质量、血糖,留取尿液、血液及肾脏。全自动特定蛋白分析仪检测UACR、尿液N-乙酰-β-D氨基葡萄糖(NAG)、尿液视黄醇结合蛋白(RBP),ELISA法检测大鼠血清炎症因子IL-1β、IL-18的水平,HE和Masson染色观察肾脏病理改变,Western blot检测肾脏组织Klotho蛋白、NF-κB蛋白的表达,免疫组化(IHC)染色观察肾脏Klotho蛋白的表达情况。结果与正常组相比,模型组大鼠体质量增加减慢(P<0.01),血糖明显升高(P<0.01),UACR、尿NAG、尿RBP明显升高(P<0.01),血清IL-1β、IL-18明显升高(P<0.01);HE染色显示,模型组大鼠肾小球体积增大,肾小管上皮细胞出现肿胀、空泡变性;Masson染色显示模型组大鼠肾小球基底膜增厚,系膜增宽,肾小管间质轻度纤维化;IHC染色显示正常组大鼠Klotho蛋白在肾小管表达明显,模型组大鼠Klotho蛋白表达下降(P<0.01);Western blot检测显示模型组大鼠肾脏组织Klotho蛋白表达明显减少(P<0.01),NF-κB蛋白表达明显增多(P<0.01)。与模型组比较,益肾康可明显升高模型大鼠体质量,降低大鼠血糖、UACR、尿NAG、尿RBP以及血清炎性因子IL-1β、IL-18水平(P<0.01),上调肾脏组织Klotho蛋白表达,下调NF-κB蛋白表达,减轻DM大鼠肾脏损伤。结论中药复方益肾康可减轻糖尿病大鼠肾脏损伤,其机制与上调Klotho蛋白表达,抑制NF-κB信号通路表达有关。Objective Based on Klotho/NF-κB signaling pathway to explore the therapeutic mechanism of Yishenkang(益肾康)in protecting diabetes kidney.Methods A diabetic rat model was established by single intraperitoneal injection of streptozotocin(53 mg/kg).Male SD rats were randomly divided into normal control group(n=10),model group(n=16),Irbesartan group(n=15)and Yishenkang group(n=15).After 4 weeks of intervention,urine was collected to detect UACR.After 8 weeks of intervention,the body weight and blood glucose of rats were measured,and urine,blood and kidney were retained.UACR and urine N-acetyl were detected by automatic specific protein analyzer-β-D-glucosamine(NAG)and urinary retinol binding protein(RBP),the level of serum inflammatory factor IL-1β,IL-18 were detected by ELISA,the pathological changes of kidney were observed by HE staining and Masson staining,the expression of Klotho and NF-κB in kidney were detected by Western blot,and the expression of Klotho protein in kidney was observed by immunohistochemistry(IHC).Results Compared with the normal group,the weight gain of model group rats slowed down(P<0.01),blood glucose increased significantly(P<0.01),UACR,urinary NAG,urinary RBP increased significantly(P<0.01),IL-1β,IL-18 increased significantly(P<0.01).HE staining showed that the glomerular volume of the model group increased,and the renal tubular epithelial cells showed swelling and vacuolar degeneration.Masson staining showed that the glomerular basement membrane of the model group thickened,mesangium widened,and renal tubulointerstitial fibrosis was mild.IHC staining showed that the expression of Klotho protein in renal tubules was obvious in normal group and decreased in model group(P<0.01).Western blot showed that the expression of Klotho protein decreased significantly(P<0.01)and the expression of NF-κB protein increased significantly(P<0.01).Compared with the model group,Yishenkang could significantly increase the body weight of model rats,reduce blood glucose,UACR,urinary NAG,urinary RBP(P<

关 键 词：糖尿病肾脏病 益肾康 补肾健脾活血法 KLOTHO NF-κB 

分 类 号：R285.5[医药卫生—中药学]