降糖三黄片对早期糖尿病肾病大鼠肾组织TGF-β1／S mad 1信号通路的影响

摘要

Objective:To observe the effect of Jiangtangsanhuang (JTSH)on growth factor-β1 (TGF-β1 ),Smad1 ,Co1-Ⅳof early dia-betic nephropathy rats,and investigate its protective effect for kidney.Methods:Eighty SD rats were randomly divided into normal control group (n=11)and diabetic model group (n=69).The diabetic models were induced by lower-dosage of streptozocin (STZ)(30mg/kg).The model group was then divided into three sub-groups including early DN group,Irbesartan group (0.027 g·kg-1 ·d-1 )and JTSH group(0.675 g·kg-1 ·d-1 ).The course of treatment lasted for 8 weeks.Rats were tested with blood glucose and weighed before dosing,4 and 8 weeks after dosing.Microalbuminuria was tested after DN model was established and before execution.Weight of all rats were measured before being executed.Immunohistochemical staining and fluorescence quota RT-PCR were used to determine smad1 and Col-Ⅳ.ELASA was used to determine TGF-β1 .Results:All the rats had weight loss after being made into disease models,and JTSH and Irbesartan groups had better effect of alleviating weight/kidney weight index (P<0.01)and 24 hours urine immunohistochemical(P<0.05)compare to early DN group.JTSH Tablet had better effect in reducing blood glucose than Irbesartan and early DN groups(P<0.01).JTSH and Irbesartan groups could significantly reduce TGF-β1(P<0.01)in rats kidney.The expression of smad1 and Col-Ⅳa-mong DN group was significantly higher than normal group by using immunohistochemical method,but there was substantial reduction in JTSH and Irbesartan groups(P<0.01).As dye index and fluorescence quota RT-PCR results,JTSH groups could reduce gene expression of smad1 and Col-Ⅳas well as Irbesartan.Conclusion:JTSH tablet has restrained TGF-β1/smad1 channel to deduce Col-Ⅳin kidney, so as to delay renal glomerulus sclerosis.%目的：观察降糖三黄片对早期糖尿病肾病大鼠肾组织中转化生长因子β1（TGF-β1）、Smad1、Co1-Ⅳ的影响，探讨其对肾脏的保护作用。方法：将80只SD大鼠随机分为正常组11只、造模组69只。腹腔内注射链脲佐菌素（STZ）复制糖尿病肾病大鼠模型。造模成功的大鼠随机进一步分为模型组、厄贝沙坦组（给药量0.027g·kg-1·d-1）、降糖三黄片组（给药量0.675g·kg-1·d-1），给药8周后处死。于治疗前、模后4周、8周各组大鼠测血糖、称体重；造模成功后、实验结束前收集24h尿液测定尿微量白蛋白。实验结束后摘取肾脏，去包膜称重；分别用免疫组化染色法及荧光定量RT-PCR法检测smad1、Ⅳ型胶原（Col-Ⅳ）；用双抗体夹心法（ELASA）检测肾组织TGF-β1。结果：造模后各组大鼠体重明显减轻，降糖三黄片、厄贝沙坦与模型组比较，肾重/体重指数下降（P＜0.01）。降糖三黄片、厄贝沙坦与模型组比较尿微量白蛋白水平下降（P＜0.05）。降糖三黄片与模型组、厄贝沙坦组相比，能较好地降低血糖（P＜0.01）；降糖三黄片、厄贝沙坦均能明显抑制糖尿病肾病大鼠肾组织中TGF-β1的表达（P＜0.01）。免疫组化染色表明厄贝沙坦组和降糖三黄片组比模型组smad1、Col-Ⅳ阳性表达程度显著减弱。免疫组化图像染色指数及荧光定量RT-PCR法结果提示降糖三黄片能降低smad1、Col-Ⅳ水平（P＜0.05），且与厄贝沙坦作用相近。结论：降糖三黄片可通过抑制肾组织TGF-β1/smad1通路的表达，降低Col-Ⅳ的沉积，使肾脏的异常结构得到改善，延缓肾小球硬化。