抗纤灵冲剂调控基因与蛋白表达干预肾间质纤维化的机制研究  

Study on Mechanism of Kangxianling Granule(抗纤灵冲剂)Intervening Renal Interstitial Fibrosis from Regulating Gene and Protein Expression

作　　者：张新志[1] 董飞侠 钱玲[3] 吴锋[1] 黄迪[4] 高建东[4] 何立群[1] ZHANG Xinzhi;DONG Feixia;QIAN Ling;WU Feng;HUANG Di;GAO Jiandong;HE Liqun(Department of Nephrology,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200021,China;Wenzhou Hospital of Chinese Medicine Affiliated to Zhejiang Chinese Medical University,Wenzhou 325000,Zhejiang,China;Shanghai East Hospital,Affiliated to Tongji University School of Medicine,Shanghai 200120,China;TCM Institute of Kidney Disease,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院,上海200021 [2]浙江中医药大学附属温州中医院,浙江温州325000 [3]同济大学附属东方医院,上海200120 [4]上海中医药大学中医肾病研究所,上海201203

出　　处：《辽宁中医杂志》2022年第1期173-176,共4页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81874437,30873259);何立群上海市名老中医学术经验研究工作室(SHGZS-2017027);上海市优秀学科带头人培养计划资助(08XD14039);肝肾疾病病证教育部重点实验室;上海市中医临床重点实验室(14DZ2273200);海派中医流派传承研究基地(ZYSNXDCC-YJXYY);上海市进一步加快中医药事业发展三年行动计划建设项目[ZY(2018-2020)-CCCX-2003-08,ZY(2018-2020)-FWTX-7005];上海市浦东新区卫生系统重点学科群建设资助(PWZxq2017-07)

摘　　要：目的从MMP-9调节COL4α3基因及Ⅳ型胶原蛋白表达的角度,研究抗纤灵冲剂干预肾间质纤维化的机制。方法72只SD大鼠,成功制备单侧输尿管梗阻(unilateral ureteral obstruction,UUO)模型后,再随机分为模型组、抗纤灵组、氯沙坦组。另24只大鼠,制备假手术模型。药物干预后处死,留取左侧肾组织。应用免疫组化法测定MMP-9、Ⅳ型胶原(Col-Ⅳ)蛋白在肾间质的阳性表达;采用PCR Array法,检测肾组织COL4α3基因的表达。结果UUO假手术组大鼠MMP-9表达极微;在模型组大鼠其阳性表达显著增加;与模型组比较,抗纤灵组4个时间点MMP-9的阳性表达均明显增加;氯沙坦组各时间点MMP-9阳性表达均较模型组增加,但无统计学差异。14d、28d时MMP-9阳性表达抗纤灵组显著高于氯沙坦组。各时间点模型组COL4α3基因水平分别较假手术组上调10.37倍、10.27倍及14.34倍。各时间点抗纤灵组COL4α3的基因表达水平,分别较模型组下调1.5倍、1.52倍和1.48倍。假手术组大鼠Col-Ⅳ只有极少量表达;模型组大鼠肾小球基底膜及肾间质表达显著增加,与假手术组相比有统计学意义;与模型组相比较,抗纤灵组及氯沙坦组各时间点阳性表达均显著减少;与氯沙坦组比较,7 d,14 d,28 d时间点Col-Ⅳ蛋白阳性表达,抗纤灵组均显著减低;21d阳性表达也明显低于氯沙坦组。结论抗纤灵冲剂干预肾间质纤维化的可能机制是通过MMP-9下调COL4α3基因及Col-Ⅳ蛋白在肾间质的表达。Objective To study the mechanism of Kangxianling Granule(抗纤灵冲剂)from MMP-9 regulating COL4α3 gene and collagen typeⅣprotein expression in renal interstitial fibrosis.Methods Seventy-two of 96 SPF Sprague-Dawley rats were ligated to prepare unilateral ureteral obstruction(UUO)rat models.After successful modeling,they were divided into model group,Kangxianling Granule(KXL)group and Losartan group.Twenty-four S-D rats rats were ligated to prepare sham-operated rat models.After intervention,we killed the rats in groups and collected the renal tissue.After that,COL4α3 gene level was detected by PCR Array method,protein expressions of MMP-9 andⅣcollagen in renal interstitium were determined by immunohistochemistry.Results The expression of MMP-9 in the rats of UUO sham group was very small.The positive expression was increased significantly in the model group.Compared with that of the model group,it was significantly increased in the KXL group at four time points.The positive expression of MMP-9 in the losartan group was increased at all time points compared with that of the model group,but there was no statistical difference.At 14 d and 28 d,the positive expression of MMP-9 in the KXL group was significantly higher than that in the losartan group.The levels of COL4-α3 gene in the model group at each time point were increased by 10.37 times,10.27 times and 14.34 times compared with those in the sham operation group,respectively more than 2 times,showing very significant differences.The expression of COL4α3 gene in KXL group was down-regulated with significant differences at each time point by 1.5 times,1.52 times and 1.48 times,respectively more than 1.5 times.Only a small amount of Col-Ⅳwas expressed in the sham group.The positive expression of glomerular basement membrane and interstitium in the model group was significantly increased by comparing with that in sham-operated group.Compared with that of the model group,the positive expressions at each time point of KXL group and Losartan group were s

关 键 词：抗纤灵冲剂 基质金属蛋白酶-9 COL4α3基因 Ⅳ型胶原 定制基因芯片 肾间质纤维化 

分 类 号：R285.5[医药卫生—中药学]