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(Nfkbp65)、环氧合酶-2(Cox2)、转化生长因子-β1(Tgfb)l)、Ⅳ型胶原(Col-Ⅳ)、α-平滑肌肌动蛋白(As-ma)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 mR-NA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。
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
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 strepto-zotocin(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 nitro-gen(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 re-nal 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 iKαb kinase(Iκkβ),nuclear transcription factor-κb p65(Nfκb p65),cyclooxygenase-2(Cox2),transforming growth factor-β1(Tgfβ1),collagen Ⅳ(ColⅣ),and α-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,thick-ened 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 expressions of Tgfβ1,ColⅣ,and α-sma(P＜0.05),and the mRNA and protein expressions of Iκkβ,Nfκb p65,and Cox2 in kidney tissue(P＜0.05).Compared with the model control group,modified Taohong Siwu Decoction(6.48 and 12.96 g/kg)reduced the 24 hUTP and BUN in DN rats,and down-regulated IL-1β,IL-6 and TNF-α(P＜0.05).They also alleviated the degree of pathological damage,and inhibited the mRNA and protein expressions of related factors in IKKβ/NF-κB/COX-2 signaling pathway as well as fibrosis-related factors(P＜0.05).Conclusion:Modified Taohong Siwu Decoction can decrease inflammatory response and fibrosis progression by inhibiting the IKKβ/NF-κB/COX-2 signaling pathway in DN rats,and relieve renal patho-logical damage,showing a clear renal protective effect.
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