氟非尼酮减缓糖尿病肾病模型小鼠肾纤维化  

Fluorofenidone reduces renal fibrosis in diabetic kidney disease mice

作　　者：谢菲菲 陆苗苗[1,2] 府晓[4] 梅文娟 雷群娟 孟婕[5] 胡高云[6] 彭张哲[1] 陶立坚[1] Xie Feifei;Lu Miaomiao;Fu Xiao;Mei Wenjuan;Lei Qunjuan;Meng Jie;Hu Gaoyun;Peng Zhangzhe;Tao Lijian(Division of Nephrology, Xiangya Hospital, Central South University, Changsha 410008, China)

机构地区：[1]中南大学湘雅医院肾内科,长沙410008 [2]辽宁医学院附属第一医院肾内科 [3]南昌医科大学附属第一医院肾内科 [4]中南大学湘雅医院血液内科,长沙410008 [5]中南大学湘雅医院呼吸内科,长沙410008 [6]中南大学药学院药物化学系

出　　处：《中华肾脏病杂志》2018年第4期288-294,共7页Chinese Journal of Nephrology

基　　金：国家自然科学基金（81470255）;国家重点研发计划（合作）（2016YFC1305505）;中华医学会临床医学科研专项资金.施维雅肾脏病青年研究与发展项目（15020090597）

摘　　要：目的观察氟非尼酮（AKF-PD）对糖尿病肾病模型db／db小鼠的疗效，并初步探讨其可能的作用机制。方法（1）14只野生型8周龄小鼠为正常对照组。42只8周龄db／db小鼠采用完全随机的方法分为模型组、AKF．PD（250mg·kg^-1·d^-1）治疗组及氯沙坦（20mg·kg^-1·d^-1）治疗组，每组14只，雌雄各半。从10周龄起治疗组分别给予相应剂量的药物连续灌胃18周，正常组及模型组给予等量药物溶媒（0．5％羧甲基纤维素钠）灌胃。尾静脉采血测血糖，ELISA测定24h尿白蛋白量，PAS染色观察各组小鼠肾脏组织病理改变，免疫组织化学检测Ⅳ型胶原及纤连蛋白（FN）在肾组织的表达。（2）体外培养小鼠。肾小球系膜细胞（MES-13），分为正常糖组（5．5mmol／L葡萄糖）、高渗组（5．5mmol／L葡萄糖＋19．5mmol／L甘露醇）、高糖组（25．0mmol／L葡萄糖）、AKF．PD组（25．0mmol／L葡萄糖＋400mg／LAKF．PD）及氯沙坦组（25．0mmol／L葡萄糖＋2μmol／L氯沙坦）。药物作用72h后，实时定量PCR检测细胞I型胶原、Ⅳ型胶原、转化生长因子B1（TGF-β1）mRNA的表达，ELISA检测细胞培养液上清中的TGF-β1蛋白含量。结果（1）与野生型小鼠比较，db／db小鼠体型肥胖，血糖明显升高，伴有大量白蛋白尿，发生肾小球肥大、系膜区扩张等病理改变，肾小球硬化指数升高（P〈0．01），肾组织Ⅳ型胶原和FN表达均增加（均P〈0.01）；与db／db小鼠比较，AKF—PD及氯沙坦治疗组小鼠24h尿白蛋白量降低（均P〈0．01），肾小球肥大及系膜区扩张减轻，肾小球硬化指数降低（均P〈0．01），肾组织中Ⅳ型胶原及FN的表达减少（均P〈0．01）。（2）与正常糖组比较，高糖组小鼠肾小球系膜细胞Ⅰ型胶原、Ⅳ型胶原mRNA表达及TGF-β1 mRNA和蛋白表达均升高（均P〈0．01）。与高糖组比较，AKF—PD组、氯沙坦组小鼠肾小球系膜�Objective To investigate the effects of fluorofenidone （AKF- PD） on diabetic kidney disease in db/db mice and its possible mechanisms. Methods （1） Fifty- six mice aged 8 weeks （half male and half female）, including 42 db/db mice and 14 wild-type mice were studied. Forty-two db/db mice randomly were divided into model group （mock-treated diabetic db/db mice）, AKF-PD （250 mg·kg^-1·d^-1） treatment group and losartan （20 mg·kg^-1·d^-1） treatment group. Wild-type mice and model mice were treated with vehicle （0.5% sodium carboxymethylcellulose）, while the treatment groups received either AKF- PD or losartan. After 18 weeks, the blood glucose and urinary albumin were measured, the pathological changes of kidney were observed by PAS staining. The protein expressions of type IV collagen and fibronectin （FN） in kidney tissue were detected by immunohistochemistry. （2） Mouse glomerular mesangial cells （MES-13 cells） were divided into six groups： normal glucose group （5.5 mmol/L glucose）, hypertonic group （5.5 mmol/L glucose ＋ 19.5 mmol/L mannitol）, high glucose group （25.0 mmol/L glucose）, AKF-PD group （25.0 mmol/L glucose＋400 mg/L AKF-PD） and losartan group （25.0 mmol/L glucose ＋ 2 μmol/L losartan）. After 72 h treatment, the expressions of type I collagen, type IV collagen and transforming growth factor-β1 （TGF-131） mRNA were detected by real- time PCR, and the content of TGF-β1 protein in the culture supernatant was detected by ELISA. Results （1） Compared with the wild type mice, model mice had increased weight, blood glucose and glomerulosclerosis index （all P 〈 0.01）, accompanied with heavy albuminuria, glomerular hypertrophy, mesangial area expansion and deposition of collagen type IV and FN （all P 〈 0.01）. Compared with model mice, in AKF- PD and losartan groups 24 h urinary albumin and glomerulosclerosis index decreased （all P 〈 0.01）, glomerular hypertrophy and mesangial area expansion alleviated, and the protein exp

关 键 词：糖尿病肾病 纤维化 转化生长因子Β1 吡啶酮类 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]