中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2013年
1期
33-38
,共6页
牛洪琳%李英%刘茂东%迟雁青%张涛%王秀芬
牛洪琳%李英%劉茂東%遲雁青%張濤%王秀芬
우홍림%리영%류무동%지안청%장도%왕수분
糖尿病肾病%系膜细胞%纤维化%整合素连接激酶%贝那普利
糖尿病腎病%繫膜細胞%纖維化%整閤素連接激酶%貝那普利
당뇨병신병%계막세포%섬유화%정합소련접격매%패나보리
Diabetic nephropathies%Mesangial cells%Fibrosis%Intergrin-linked kinase%Benazepril
目的 观察贝那普利对高糖培养下肾小球系膜细胞(GMC)中整合素连接激酶(ILK)及α平滑肌肌动蛋白(α-SMA)表达的影响,探讨贝那普利肾脏保护作用的调节机制.方法 将GMC常规培养分为4组:正常糖对照组(D-葡萄糖5.5 mmol/L,NG组)、甘露醇组(甘露醇20 mmol/L,MG组)、高糖组(D-葡萄糖30 mmol/L,HG组)、高糖+贝那普利干预组(D-葡萄糖30 mmol/L+贝那普利10 μmol/L,ACEI组).于实验开始后3h、6h、12h、24 h、48 h、72 h分别收获4组细胞,应用RT-PCR法、Western印迹法和细胞免疫荧光法检测各组GMC中ILK、α-SMA的表达.结果 高糖环境下GMC中ILK、α-SMA mRNA和蛋白表达水平较正常对照组显著升高(均P＜ 0.05),ILK在48 h达到最高(P＜0.05),α-SMA在72 h达到最高(P＜0.01).贝那普利干预后上述指标较高糖组表达水平下降,但未能恢复至正常糖对照组水平.同时间点甘露醇组ILK mRNA和蛋白水平与正常糖组差异无统计学意义(P＞0.05)；同时间点甘露醇组α-SMA mRNA和蛋白水平显著高于正常糖对照组(均P＜0.05).结论 贝那普利可能通过降低ILK和α-SMA的异常表达来延缓糖尿病肾病的肾脏纤维化进程和高糖介导的系膜细胞表型转化.系膜细胞在糖尿病肾病中的表型转化可能同时存在渗透浓度依赖性.
目的 觀察貝那普利對高糖培養下腎小毬繫膜細胞(GMC)中整閤素連接激酶(ILK)及α平滑肌肌動蛋白(α-SMA)錶達的影響,探討貝那普利腎髒保護作用的調節機製.方法 將GMC常規培養分為4組:正常糖對照組(D-葡萄糖5.5 mmol/L,NG組)、甘露醇組(甘露醇20 mmol/L,MG組)、高糖組(D-葡萄糖30 mmol/L,HG組)、高糖+貝那普利榦預組(D-葡萄糖30 mmol/L+貝那普利10 μmol/L,ACEI組).于實驗開始後3h、6h、12h、24 h、48 h、72 h分彆收穫4組細胞,應用RT-PCR法、Western印跡法和細胞免疫熒光法檢測各組GMC中ILK、α-SMA的錶達.結果 高糖環境下GMC中ILK、α-SMA mRNA和蛋白錶達水平較正常對照組顯著升高(均P＜ 0.05),ILK在48 h達到最高(P＜0.05),α-SMA在72 h達到最高(P＜0.01).貝那普利榦預後上述指標較高糖組錶達水平下降,但未能恢複至正常糖對照組水平.同時間點甘露醇組ILK mRNA和蛋白水平與正常糖組差異無統計學意義(P＞0.05)；同時間點甘露醇組α-SMA mRNA和蛋白水平顯著高于正常糖對照組(均P＜0.05).結論 貝那普利可能通過降低ILK和α-SMA的異常錶達來延緩糖尿病腎病的腎髒纖維化進程和高糖介導的繫膜細胞錶型轉化.繫膜細胞在糖尿病腎病中的錶型轉化可能同時存在滲透濃度依賴性.
목적 관찰패나보리대고당배양하신소구계막세포(GMC)중정합소련접격매(ILK)급α평활기기동단백(α-SMA)표체적영향,탐토패나보리신장보호작용적조절궤제.방법 장GMC상규배양분위4조:정상당대조조(D-포도당5.5 mmol/L,NG조)、감로순조(감로순20 mmol/L,MG조)、고당조(D-포도당30 mmol/L,HG조)、고당+패나보리간예조(D-포도당30 mmol/L+패나보리10 μmol/L,ACEI조).우실험개시후3h、6h、12h、24 h、48 h、72 h분별수획4조세포,응용RT-PCR법、Western인적법화세포면역형광법검측각조GMC중ILK、α-SMA적표체.결과 고당배경하GMC중ILK、α-SMA mRNA화단백표체수평교정상대조조현저승고(균P＜ 0.05),ILK재48 h체도최고(P＜0.05),α-SMA재72 h체도최고(P＜0.01).패나보리간예후상술지표교고당조표체수평하강,단미능회복지정상당대조조수평.동시간점감로순조ILK mRNA화단백수평여정상당조차이무통계학의의(P＞0.05)；동시간점감로순조α-SMA mRNA화단백수평현저고우정상당대조조(균P＜0.05).결론 패나보리가능통과강저ILK화α-SMA적이상표체래연완당뇨병신병적신장섬유화진정화고당개도적계막세포표형전화.계막세포재당뇨병신병중적표형전화가능동시존재삼투농도의뢰성.
Objective To investigate the effect of benazepril on intergrin-linked kinase (ILK) and α-smooth muscle actin (α-SMA) expression in glomerular mesangial cells induced by high-glucose.Methods The mesangial cells from SD rat (HBZY-1) were cultured conventionally and randomly divided into four groups:normal glucose (D-glucose 5.5 mmol/L,group NG),mannitol-treated group (mannitol 20 mmol/L,group MG),high glucose (D-glucose 30 mmol/L,group HG),Benazepril-treated high glucose group (D-glucose 30 mmol/L + Benazepril 10 μmol/L,group ACEI).Cells from NG,MG,HG,ACEI gronps were harvested after 3,6,12,24,48 and 72 hours of treatment respectively.The mRNA expressions of ILK and α-SMA were detected by RT-PCR.The protein levels of ILK and α-SMA were detected by Western blotting and immunofluorescence.Results The expressions of ILK mRNA and protein in HG group were significantly increased compared with those in NG group (all P ＜ 0.05).The increased expressions of ILK and α-SMA in HG group were time-dependent and the expression reached the peak at 48 h (ILK,P ＜ 0.05) or 72 h (α-SMA,P ＜ 0.01).The expressions of ILK and α-SMA in ACEI group were lower than those in HG group (all P ＜ 0.01),but failed to rescue to the same level as those in NG.There was no significant differences of ILK expressions between MG group and NG group at the same time point (P ＞ 0.05).The expressions of α-SMA mRNA and protein in MG were higher than that in NG (P ＜ 0.05),which suggest that high osmotic pressure could cause the increasing of α-SMA.Conclusions Benazepril can decrease the expressions of ILK and α-SMA to inhibit the process of fibrosis in DN and mediate the phenotypic transformation of glomerular mesangial cells.The phenotypic transformation of glomerular mesangial cells in glucose may also depend on high osmotic pressure in DN.
