中南大学学报（医学版）
中南大學學報（醫學版）
중남대학학보（의학판）
JOURNAL OF CENTRAL SOUTH UNIVERSITY (MEDICAL SCIENCES)
2013年
12期
1233-1242
,共10页
罗俊辉%李瑛%杨阳%李军%孙林%段绍斌%刘虹%刘伏友%刘玉平%奚易云%尤燕华%李华
囉俊輝%李瑛%楊暘%李軍%孫林%段紹斌%劉虹%劉伏友%劉玉平%奚易雲%尤燕華%李華
라준휘%리영%양양%리군%손림%단소빈%류홍%류복우%류옥평%해역운%우연화%리화
曲尼斯特%糖尿病肾病%肾间质纤维化%肥大细胞%干细胞生长因子%干细胞因子受体
麯尼斯特%糖尿病腎病%腎間質纖維化%肥大細胞%榦細胞生長因子%榦細胞因子受體
곡니사특%당뇨병신병%신간질섬유화%비대세포%간세포생장인자%간세포인자수체
tranilast%diabetic kidney disease%tubulointerstitial ifbrosis%mast cells%stem cell factor%c-kit
目的：研究曲尼斯特延缓糖尿病肾病肾间质纤维化的作用及机制。方法：建立糖尿病肾病(DKD)大鼠模型：SD大鼠随机分为正常对照组(n=6)、DKD模型组(n=8)、曲尼斯特低剂量(n=8)和高剂量治疗组(n=8)。采用高糖高脂饲料喂养联合低剂量STZ注射构建大鼠DKD模型。成模后，分别予以曲尼斯特200 mg/(kg·d)(曲尼斯特低剂量组)和400 mg/(kg·d)(曲尼斯特高剂量组)分2次灌胃。于第8周末处死大鼠，收集大鼠24 h尿液测24 h尿白蛋白排泄量，收集血测肾功能及血白蛋白；取部分肾组织置于4%中性甲醛溶液中固定，采用免疫组织化学检测肾组织补体C3a受体(C3aR)，E-钙黏附蛋白(epithelial cadherin，E-Cadherin)，α-SMA，纤维连接蛋白(fibronectin，FN)，I型胶原蛋白(collagen I，Col I)，干细胞生长因子(stem cell factor， SCF)和干细胞因子受体(c-kit)的表达以及分布；Western印迹检测肾组织E-cadherin，α-SMA，FN，Col I，SCF和c-kit蛋白的表达；RT-PCR检测肾组织FN，Col I，SCF，c-kit mRNA的表达。结果：曲尼斯特能抑制肥大细胞在DKD大鼠肾组织的浸润；DKD模型组肾小管上皮细胞E-cadherin的表达较正常对照组减少，并可见α-SMA表达，曲尼斯特可一定程度逆转这一过程；与正常对照组比较，DKD模型组肾小管间质区域Col I和FN的表达增加，曲尼斯特能剂量依赖性地抑制Col I和FN的表达；DKD大鼠肾组织SCF，c-kit蛋白及mRNA表达增加；肾组织SCF，c-kit蛋白表达与肥大细胞浸润程度及肾小管间质FN，Col I蛋白的表达呈显著正相关。曲尼斯特能抑制SCF，c-kit mRNA及蛋白的表达(P<0.05)。结论：肥大细胞参与了DKD大鼠肾间质纤维化的发生发展，曲尼斯特可能通过阻断SCF/c-kit信号通路，抑制肥大细胞的募集而逆转DKD大鼠肾小管上皮细胞的EMT，抑制肾间质纤维化。
目的：研究麯尼斯特延緩糖尿病腎病腎間質纖維化的作用及機製。方法：建立糖尿病腎病(DKD)大鼠模型：SD大鼠隨機分為正常對照組(n=6)、DKD模型組(n=8)、麯尼斯特低劑量(n=8)和高劑量治療組(n=8)。採用高糖高脂飼料餵養聯閤低劑量STZ註射構建大鼠DKD模型。成模後，分彆予以麯尼斯特200 mg/(kg·d)(麯尼斯特低劑量組)和400 mg/(kg·d)(麯尼斯特高劑量組)分2次灌胃。于第8週末處死大鼠，收集大鼠24 h尿液測24 h尿白蛋白排洩量，收集血測腎功能及血白蛋白；取部分腎組織置于4%中性甲醛溶液中固定，採用免疫組織化學檢測腎組織補體C3a受體(C3aR)，E-鈣黏附蛋白(epithelial cadherin，E-Cadherin)，α-SMA，纖維連接蛋白(fibronectin，FN)，I型膠原蛋白(collagen I，Col I)，榦細胞生長因子(stem cell factor， SCF)和榦細胞因子受體(c-kit)的錶達以及分佈；Western印跡檢測腎組織E-cadherin，α-SMA，FN，Col I，SCF和c-kit蛋白的錶達；RT-PCR檢測腎組織FN，Col I，SCF，c-kit mRNA的錶達。結果：麯尼斯特能抑製肥大細胞在DKD大鼠腎組織的浸潤；DKD模型組腎小管上皮細胞E-cadherin的錶達較正常對照組減少，併可見α-SMA錶達，麯尼斯特可一定程度逆轉這一過程；與正常對照組比較，DKD模型組腎小管間質區域Col I和FN的錶達增加，麯尼斯特能劑量依賴性地抑製Col I和FN的錶達；DKD大鼠腎組織SCF，c-kit蛋白及mRNA錶達增加；腎組織SCF，c-kit蛋白錶達與肥大細胞浸潤程度及腎小管間質FN，Col I蛋白的錶達呈顯著正相關。麯尼斯特能抑製SCF，c-kit mRNA及蛋白的錶達(P<0.05)。結論：肥大細胞參與瞭DKD大鼠腎間質纖維化的髮生髮展，麯尼斯特可能通過阻斷SCF/c-kit信號通路，抑製肥大細胞的募集而逆轉DKD大鼠腎小管上皮細胞的EMT，抑製腎間質纖維化。
목적：연구곡니사특연완당뇨병신병신간질섬유화적작용급궤제。방법：건립당뇨병신병(DKD)대서모형：SD대서수궤분위정상대조조(n=6)、DKD모형조(n=8)、곡니사특저제량(n=8)화고제량치료조(n=8)。채용고당고지사료위양연합저제량STZ주사구건대서DKD모형。성모후，분별여이곡니사특200 mg/(kg·d)(곡니사특저제량조)화400 mg/(kg·d)(곡니사특고제량조)분2차관위。우제8주말처사대서，수집대서24 h뇨액측24 h뇨백단백배설량，수집혈측신공능급혈백단백；취부분신조직치우4%중성갑철용액중고정，채용면역조직화학검측신조직보체C3a수체(C3aR)，E-개점부단백(epithelial cadherin，E-Cadherin)，α-SMA，섬유련접단백(fibronectin，FN)，I형효원단백(collagen I，Col I)，간세포생장인자(stem cell factor， SCF)화간세포인자수체(c-kit)적표체이급분포；Western인적검측신조직E-cadherin，α-SMA，FN，Col I，SCF화c-kit단백적표체；RT-PCR검측신조직FN，Col I，SCF，c-kit mRNA적표체。결과：곡니사특능억제비대세포재DKD대서신조직적침윤；DKD모형조신소관상피세포E-cadherin적표체교정상대조조감소，병가견α-SMA표체，곡니사특가일정정도역전저일과정；여정상대조조비교，DKD모형조신소관간질구역Col I화FN적표체증가，곡니사특능제량의뢰성지억제Col I화FN적표체；DKD대서신조직SCF，c-kit단백급mRNA표체증가；신조직SCF，c-kit단백표체여비대세포침윤정도급신소관간질FN，Col I단백적표체정현저정상관。곡니사특능억제SCF，c-kit mRNA급단백적표체(P<0.05)。결론：비대세포삼여료DKD대서신간질섬유화적발생발전，곡니사특가능통과조단SCF/c-kit신호통로，억제비대세포적모집이역전DKD대서신소관상피세포적EMT，억제신간질섬유화。
Objective:To determine the role and mechanism of tranilast preventing the progression of tubulointerstilial ifbrosis in diabetic kidney disease (DKD). <br> Methods:Sprague-Dawley rats were randomly divided into a control group (n=6), DKD model group (n=8), low dose tranilast group [200 mg/(kg.d), n=8], and high dose tranilast group [400 mg/(kg.d), n=8]. Tranilast was administered daily after the model was built. Rats were sacrificed at day 56, 24 hour urine was collected to measure 24-hour urine albumin excretion, and blood was collected to determine the renal function and serum albumin. Then the kidneys were harvested and subjected to studies. The expression of C3aR, E-cadherin,α-SMA, fibronectin(FN), collagen I (Col I), stem cell factor (SCF) and c-kit were detected by immunohistochemical staining respectively. The expression of E-cadherin,α-SMA, FN, Col I, SCF and c-kit protein was analyzed by Western blot, and the expression of FN, Col I, SCF and c-kit mRNA was examined by RT-PCR. Results:Tranilast can inhibit the inifltration of mast cells in the kidneys of DKD rats. The expression ofα-SMA in the kidneys of DKD rats inereased signiifcantly (P<0.05), while the expression of E-cadherin decreased (P<0.05). Tranilast increased the expression of E-cadherin and decreased the expression ofα-SMA in the prophase of DKD dose dependently. The expressions of FN and Col I were increased in the tubulointerstitial ifelds in DKD model rats (P<0.05). After the tranilast treatment, these changes were relieved to a certein degree (P<0.05). The expression of SCF and c-kit in the tubular and interstitial tissue was slight. The increased expressions of SCF and c-kit protein and mRNA in DKD model rats were downregulated by tranilat (P<0.05). The expressions of SCF and c-kit were positively correlated with the infiltration degree of mast cells and the expressions of FN, Col I. <br> Conclusion:Mast cells participate in and aggravate the renal tubulointerstitial fibrosis in DKD rats. Tranilast can reverse the EMT of renal tubular cells and inhibit the tubulointersitial fibrosis of DKD by blocking the inifltration of mast cells induced by SCF/c-kit pathway.
