基于IKKβ/NF-κB/COX-2信号通路探讨桃红四物汤加味对糖尿病肾病大鼠炎性因子及肾脏纤维化的干预作用  

Intervention Effect of Modified Taohong Siwu Decoction(桃红四物汤)on Inflammatory Factors and Renal Fibrosis in Rats with Diabetic Nephropathy Based on IKKβ/NF-κB/COX-2 Signaling Pathway

作　　者：白璐 陈永哲 王倩[1,2] 李敏 沈士朋 郭帅 杨帆[3] 刘茂东 BAI Lu;CHEN Yongzhe;WANG Qian;LI Min;SHEN Shipeng;GUO Shuai;YANG Fan;LIU Maodong(Department of Nephrology,Hebei Medical University Third Hospital,Shijiazhuang 050051;Hebei Provincial Key Laboratory of Diabetic Kidney Disease,Shijiazhuang 050051;Hebei University of Chinese Medicine,Shijiazhuang 050091)

机构地区：[1]河北医科大学第三医院肾内科,石家庄050051 [2]河北省糖尿病肾脏疾病重点实验室,石家庄050051 [3]河北中医药大学,石家庄050091

出　　处：《中药药理与临床》2024年第6期11-17,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：河北省中医药管理局科研计划项目(编号:2019131、2022408);河北省自然科学基金项目(编号:H2022423320)。

摘　　要：目的:探讨桃红四物汤加味对糖尿病肾病大鼠炎性因子及肾脏IKKβ/NF-κB/COX-2信号通路表达及纤维化的影响。方法:10只大鼠随机选为正常对照组,其余大鼠采用高脂饲料联合链脲佐菌素35 mg/kg腹腔注射建立糖尿病肾病模型,模型制备成功后分为模型对照组、厄贝沙坦0.0135 g/kg组、桃红四物汤加味3.24、6.48、12.96 g/kg组。各组大鼠灌胃给予相应药物干预16 w,于16 w末检测大鼠空腹血糖(FBG)、24 h尿蛋白定量(24 h UTP)、肌酐(Scr)、尿素氮(BUN)含量;ELISA法检测血浆中炎性因子白介素-1β(IL-1β)、IL-6、TNF-α含量;HE、Masson染色观察肾脏病理学改变;Real-time PCR法检测肾组织IκαB激酶β(Ikkb)、核转录因子-κB p65(Nfkb p65)、环氧合酶-2(Cox2)、转化生长因子-β1(Tgfb1)、IV型胶原(Col-IV)、α-平滑肌肌动蛋白(Asma)mRNA的表达;免疫组化法(IHC)检测肾组织TGF-β1、COL-IV、α-SMA、NF-κB p65蛋白的表达;Western blotting法检测肾组织IKKβ、NF-κB p65、COX-2蛋白的表达。结果:与正常对照组比较,模型对照组大鼠FBG、24 h UTP、BUN含量明显升高(P<0.05),光镜观察发现肾小球肥大,系膜基质增加,基底膜增厚,肾小囊囊腔变窄,肾小管部分空泡变性,血浆中IL-1β、IL-6、TNF-α含量明显增加(P<0.05),肾组织纤维化相关蛋白Tgfb1、Col-IV、Asma mRNA及蛋白表达均明显上调(P<0.05),肾组织Ikkb、Nfkb p65、Cox2 mRNA及蛋白表达明显上调(P<0.05);与模型对照组比较,桃红四物汤加味6.48、12.96 g/kg组DN大鼠24 h UTP、BUN含量降低,IL-1β、IL-6、TNF-α含量降低(P<0.05),病理损伤程度减轻,同时IKKβ/NF-κB/COX-2信号通路及纤维化相关因子mRNA和蛋白表达下调(P<0.05)。结论:桃红四物汤加味可能通过抑制DN大鼠肾脏IKKβ/NF-κB/COX-2信号通路表达,减轻炎症反应及纤维化进展,减轻肾脏病理损伤,具有明确的肾保护作用。Objective:To investigate the effect of modified Taohong Siwu Decoction(桃红四物汤)on inflammatory factors and renal fibrosis in rats with diabetic nephropathy(DN)based on IKKβ/NF-κB/COX-2 signaling pathway.Methods:Ten rats were randomly selected as normal control group,and the remaining rats were established as DN models by feeding high-fat diet combined with intraperitoneal injection of streptozotocin(35 mg/kg).After successful preparation,the modeled rats were divided into model control group,irbesartan group(0.0135 g/kg),and modified Taohong Siwu Decoction(3.24,6.48 and 12.96 g/kg).Rats in each group were given corresponding medicine by gavage for 16 weeks,and fasting blood glucose(FBG),24-hour urine protein quantification(24 hUTP),serum creatinine(Scr),and blood urea nitrogen(BUN)were detected at the end of 16 weeks.The plasma expressions of inflammatory factors interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)were measured by enzyme-linked immunosorbent assay(ELISA).The pathological changes in renal tissues were observed by HE staining and MASSON staining.Real-time polymerase chain reaction(Real-time PCR)was used to measure the mRNA expressions of iκαb kinase(Iκkβ),nuclear transcription factor-κb p65(Nfκb p65),cyclooxygenase-2(Cox2),transforming growth factor-β1(Tgfβ1),collagen IV(ColIV),andα-smooth muscle actin(α-sma).Immunohistochemistry(IHC)was employed to detect the protein expressions of TGF-β1,COL-IV,α-SMA and,NF-κB p65 in kidney tissue,and Western blotting was performed to determine the protein expressions of IKKβ,NF-κB p65 and COX-2 in kidney tissue.Results:Compared with the normal control group,the model control group had elevated FBG,24hUTP and BUN(P<0.05).Light microscopy revealed glomerular hypertrophy,increased mesangial matrix,thickened basement membrane,narrowing of renal capsule and partial vacuolar degeneration of renal tubules in the model control group,and there was an increase in serum IL-1β,IL-6 and TNF-α(P<0.05),the mRNA and protein

关 键 词：桃红四物汤加味 糖尿病肾病 炎性因子 纤维化 IκB激酶β/核因子κB/环氧酶2信号通路 

分 类 号：R285.5[医药卫生—中药学]