摘要
目的基于转化生长因子-β1(TGF-β1)/Smads通路探讨女贞子酒蒸前后改善糖尿病肾病(DN)大鼠肾纤维化的作用与机制。方法随机选取8只SD雄性大鼠作为空白组,其余大鼠高脂饲料喂养联合腹腔注射1%链脲佐菌素(65 mg/kg)建立DN模型。DN模型大鼠随机分为模型组、二甲双胍阳性药组,女贞子低、高剂量组,酒女贞子低、高剂量组,每组8只。各给药组大鼠灌胃相应药物,空白组和模型组大鼠灌胃等体积蒸馏水,1次/d,连续8周。血糖仪检测空腹血糖,考马斯亮蓝法检测24 h尿蛋白量,ELISA法测定血清中肌酐(Scr)、尿素氮(BUN)水平,Masson染色法观察大鼠肾组织病理变化,qRT-PCR法测定肾组织TGF-β1、Smad 3、Smad 7 mRNA表达水平,Western blotting法测定肾组织TGF-β1、Smad 3、磷酸化Smad 3(p-Smad 3)、Smad 7、磷酸化Smad 7(p-Smad 7)蛋白表达水平。结果与空白组相比,模型组大鼠血糖、24 h尿蛋白及血清中Scr、BUN水平显著升高(P<0.01);肾组织可见肾小球周围有胶原纤维增生,中度纤维化,系膜基质重度增生,基底膜增厚,肾小球萎缩;肾组织TGF-β1、Smad 3 mRNA表达及TGF-β1、Smad 3和p-Smad 3蛋白表达明显上调(P<0.01),肾组织Smad 7 mRNA表达及Smad 7、p-Smad 7蛋白表达明显下调(P<0.01);与模型组相比,女贞子酒蒸前后大鼠肾纤维化及肾损伤均有不同程度改善,血糖、24 h尿蛋白及血清BUN、SCr水平显著降低(P<0.05或P<0.01),肾组织TGF-β1、Smad 3 mRNA表达及TGF-β1、Smad 3、p-Smad 3蛋白表达水平明显下调(P<0.05或P<0.01),肾组织Smad 7 mRNA表达及Smad 7和p-Smad 7蛋白表达水平显著升高(P<0.05或P<0.01),且酒女贞子的改善效果优于女贞子。结论女贞子酒蒸后抗DN大鼠肾纤维化作用增强,可能是通过调控TGF-β1/Smads通路增强对DN大鼠肾纤维化的治疗作用。
Objective To study the effect and mechanism of Ligustri Lucidi Fructus before after steaming with wine on renal fibrosis in rats with diabetic nephropathy based on transforming growth factor-β1(TGF-β1)/Smads pathway.Methods Eight male SD rats were randomly selected as control group,and the rest rats were fed with high-fat diet and intraperitoneal injection of 1%streptozotocin(65 mg/kg)to establish a diabetic nephropathy(DN)model.DN model rats were randomly divided into model group,metformin group,low and high dose of Ligustri Lucidi Fructus group,low and high dose of wine Ligustri Lucidi Fructus group,with 8 rats in each group.The corresponding drugs were given to the rats in each administration group,rats in control group and model group were given the same volume of distilled water,once a day for 8 weeks.Fasting blood glucose was detected by blood glucose meter,24 h urine protein was detected by coomassie brilliant blue method,serum creatinine(Scr)and blood urea nitrogen(BUN)levels were determined by ELISA.Masson staining was used to observe the pathological situation of renal tissue.qRT-PCR was used to determine the mRNA expression of TGF-β1,Smad 3,and Smad 7 in renal tissue.Western blotting was used to determine the expression of TGF-β1,Smad 3,phosphorylated Smad 3(p-Smad 3),Smad 7 and phosphorylated Smad 7(p-Smad 7)in renal tissue.Results Compared with the control group,the blood glucose,24 h urine protein,Scr and BUN levels in the model group were significantly increased(P<0.01).Renal tissue showed collagen fiber proliferation and moderate fibrosis around the glomeruli,severe mesangial matrix proliferation,basement membrane thickening,and glomerular atrophy.TGF-β1 and Smad 3 mRNA levels and the protein expressions of TGF-β1,Smad 3 and p-Smad 3 in renal tissue were significantly increased(P<0.01),Smad 7 mRNA level and the protein expressions of Smad 7 and p-Smad 7 in renal tissue were significantly reduced(P<0.01).Compared with model group,the renal fibrosis and renal injury of rats were improved to va
作者
庄梓鸣
李钦青
蔡梅超
赵梦
孔庆悦
张学兰
栾茹乔
ZHUANG Zi-ming;LI Qin-qing;CAI Mei-chao;ZHAO Meng;KONG Qing-yue;ZHANG Xue-lan;LUAN Ru-qiao(College of Pharmacy,Shandong University of Traditional Chinese Medicine,Ji'nan Shandong 250355,China;School of Chinese Medicine and Food Engineering,Shanxi University of Chinese Medicine,Jinzhong Shanxi 030619,China)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2024年第6期1394-1398,共5页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金面上项目(81973486,82173974)
国家中医药管理局中药炮制技术传承地项目(国中医药科技中药[2022]59号)
国家重点研发计划课题(2018YFC1707002)。
