石斛合剂对糖尿病肾病小鼠TNF-α及IL-6表达的影响  

Effect of Shihu Compound on Expression of TNF-α and IL-6 in Mice with Diabetic Nephropathy

作　　者：陈勇[1] 张捷平[1] 余文珍[1] 林心君[1] 林晓晖[1] 施红[1] CHEN Yong;ZHANG Jie-ping;YU Wen-zhen;LIN Xin-jun;LIN Xiao-hui;SHI Hong(College of Integration of Traditional Chinese and Western Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China)

机构地区：[1]福建中医药大学中西医结合学院

出　　处：《中国实验方剂学杂志》2018年第18期107-112,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金青年基金项目(81503438);福建省自然科学基金项目(2018J01873);福建省教育厅中青年教师教育科研项目(JAT160230);福建中医药大学科研平台(中西医结合基础综合实验室)校管课题(X2017016-平台)

摘　　要：目的：观察石斛合剂对db/db糖尿病小鼠肾组织肿瘤坏死因子-α（TNF-α）,白细胞介素-6（IL-6）表达水平的影响。方法：选用11~12周龄雄性db/db小鼠,按空腹血糖及体质量随机分成模型组、格列吡嗪治疗组、石斛合剂低剂量组和高剂量组,db/m小鼠为正常组。连续灌胃给药8周。检测小鼠体质量及24 h尿蛋白定量;苏木素-伊红（HE）染色观察肾组织病理形态改变;通过酶联免疫吸附测定（ELISA）检测小鼠血清及肾组织TNF-α,IL-6的含量;分别应用免疫组织化学染色和蛋白免疫印迹法（Western bolt）检测肾组织中TNF-α,IL-6蛋白的表达。结果：石斛合剂能明显降低db/db小鼠的体质量（P〈0.05,P〈0.01）及24 h尿蛋白定量（P〈0.05,P〈0.01）。HE染色结果显示模型组小鼠肾小球体积增大,系膜基质增多,系膜区增宽,肾小管上皮细胞空泡变性;与模型组相比,各治疗组肾小球基底膜变薄,系膜细胞增生情况减轻,细胞外基质减少,肾小管结构基本恢复正常。免疫组织化学,ELISA和Western bolt结果表明,与正常组比较,模型组小鼠TNF-α,IL-6的表达均显著增加（P〈0.01）;与模型组比较,各治疗组TNF-α,IL-6的表达均明显减少（P〈0.05,P〈0.01）,但仍高于正常组。结论：石斛合剂可能通过下调TNF-α及IL-6的表达减轻肾组织内的炎症反应,改善db/db小鼠的肾脏损害,从而延缓DN的进展。Objective： To observe the effect of Shihu compound（ SC） on expression levels of tumor necrosis factor-alpha（ TNF-α） and interleukin-6（ IL-6） in renal tissues of db/db diabetic mice. Method： The11-12 week old male db/db mice,were randomly divided into model group,glipizide treatment group,low and high dose SC groups,and db/m normal control group according to fasting blood glucose and body mass. Intragastric administration was given for 8 weeks. Body weight and 24 hours urinary protein quantitation in mice were detected;the pathological changes of renal tissues were observed by optical microscope. Enzyme-linked immunosorbent assay（ ELISA） was used to detect the levels of TNF-α and IL-6 in serum and kidney tissue of mice;immunohistochemical staining and Western bolt were used to detect the mRNA and protein expression levels of TNF-α and IL-6 in renal tissue. Result： SC can significantly reduce the body weight（ P〈0. 05,P〈0. 01） and24 h urinary protein（ P〈0. 05,P〈0. 01） in db/db mice. Hematoxylin and eosin（ HE） staining results showed that the volume of glomerulus was increased,mesangial matrix was increased,mesangial area was widened and vacuolar degeneration of renal tubular epithelial cells was obvious in model group. As compared with the model group,the glomerular basement membrane was thinner,the proliferation of mesangial cells was decreased,the extracellular matrix was decreased,and the renal tubule structure was basically recovered to a normal condition.The results of immunohistochemistry,ELISA and Western bolt showed that the expression levels of TNF-α and IL-6 in the model group were increased significantly as compared with the normal group（ P〈0. 01）; as compared with the model group,the expression levels of TNF-α and IL-6 in each treatment group were significantly decreased（ P〈0. 05,P〈0. 01）,but still higher than those in the normal group. Conclusion： SC can alleviate the inflammatory reaction in kidney tissues and improve the renal damage in db

关 键 词：石斛合剂 DB/DB小鼠 肿瘤坏死因子-Α 白细胞介素-6 

分 类 号：R22[医药卫生—中医基础理论] R24[医药卫生—中医学]