中华眼底病杂志
中華眼底病雜誌
중화안저병잡지
CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
2015年
1期
52-56
,共5页
马海智%张少冲%黄雄高%魏雁涛%蒋欣桐
馬海智%張少遲%黃雄高%魏雁濤%蔣訢桐
마해지%장소충%황웅고%위안도%장흔동
视网膜色素上皮%细胞转分化%转化生长因子β%rac1 GTP结合蛋白质
視網膜色素上皮%細胞轉分化%轉化生長因子β%rac1 GTP結閤蛋白質
시망막색소상피%세포전분화%전화생장인자β%rac1 GTP결합단백질
Retinal pigment epithelium%Cell transdifferentiation%Transforming growth factor beta%rac1 GTP-binding protein
目的 观察转化生长因子β1(TGF-β1)诱导视网膜色素上皮(RPE)细胞间质样转分化(EMT)过程中Rac1在其细胞学行为改变中的作用.方法 人RPE-19细胞分为空白组、TGF-β1组、TGF-β1+NSC23766组、NSC23766组.所有实验于加入TGF-β1前2h给予NSC23766以封闭Rac1受体.免疫荧光、蛋白免疫印迹法检测细胞中α-平滑肌肌动蛋白(α-SMA)表达;细胞划痕、侵袭及凝胶收缩实验,观察NSC23766对细胞迁徙、侵袭、凝胶收缩作用.结果 TGF-β1组细胞中α-SMA荧光表达较空白组、TGF-β1+ NSC23766增强,差异有统计学意义(F=825.314,P=0.003);细胞划痕实验结果显示,TGF-β1组细胞间距小于TGF-β1+ NSC23766组,差异有统计学意义(P24h=0.04,P48h =0.001);与空白组细胞间距比较,差异无统计学差异(P=0.278).细胞侵袭实验结果显示,TGF-β1组穿过纤维膜的细胞数与空白组、TGF-β1+ NSC23766组、NSC23766组穿过纤维膜的细胞数比较,差异无统计学意义(F=0.371,P=0.055).凝胶收缩实验结果显示,TGF-β1组能明显增加细胞的促凝胶收缩能力,组间差异有统计学意义(F=40.473,P=0.014),TGF-β1组相对TGF-β1+ NSC23766组,差异有统计学意义(P=0.022).结论 Rac1在TGF-β1诱导RPE细胞凝胶收缩改变中发挥作用;NSC23766可通过调控Rac1活化抑制RPE细胞学行为的改变.
目的 觀察轉化生長因子β1(TGF-β1)誘導視網膜色素上皮(RPE)細胞間質樣轉分化(EMT)過程中Rac1在其細胞學行為改變中的作用.方法 人RPE-19細胞分為空白組、TGF-β1組、TGF-β1+NSC23766組、NSC23766組.所有實驗于加入TGF-β1前2h給予NSC23766以封閉Rac1受體.免疫熒光、蛋白免疫印跡法檢測細胞中α-平滑肌肌動蛋白(α-SMA)錶達;細胞劃痕、侵襲及凝膠收縮實驗,觀察NSC23766對細胞遷徙、侵襲、凝膠收縮作用.結果 TGF-β1組細胞中α-SMA熒光錶達較空白組、TGF-β1+ NSC23766增彊,差異有統計學意義(F=825.314,P=0.003);細胞劃痕實驗結果顯示,TGF-β1組細胞間距小于TGF-β1+ NSC23766組,差異有統計學意義(P24h=0.04,P48h =0.001);與空白組細胞間距比較,差異無統計學差異(P=0.278).細胞侵襲實驗結果顯示,TGF-β1組穿過纖維膜的細胞數與空白組、TGF-β1+ NSC23766組、NSC23766組穿過纖維膜的細胞數比較,差異無統計學意義(F=0.371,P=0.055).凝膠收縮實驗結果顯示,TGF-β1組能明顯增加細胞的促凝膠收縮能力,組間差異有統計學意義(F=40.473,P=0.014),TGF-β1組相對TGF-β1+ NSC23766組,差異有統計學意義(P=0.022).結論 Rac1在TGF-β1誘導RPE細胞凝膠收縮改變中髮揮作用;NSC23766可通過調控Rac1活化抑製RPE細胞學行為的改變.
목적 관찰전화생장인자β1(TGF-β1)유도시망막색소상피(RPE)세포간질양전분화(EMT)과정중Rac1재기세포학행위개변중적작용.방법 인RPE-19세포분위공백조、TGF-β1조、TGF-β1+NSC23766조、NSC23766조.소유실험우가입TGF-β1전2h급여NSC23766이봉폐Rac1수체.면역형광、단백면역인적법검측세포중α-평활기기동단백(α-SMA)표체;세포화흔、침습급응효수축실험,관찰NSC23766대세포천사、침습、응효수축작용.결과 TGF-β1조세포중α-SMA형광표체교공백조、TGF-β1+ NSC23766증강,차이유통계학의의(F=825.314,P=0.003);세포화흔실험결과현시,TGF-β1조세포간거소우TGF-β1+ NSC23766조,차이유통계학의의(P24h=0.04,P48h =0.001);여공백조세포간거비교,차이무통계학차이(P=0.278).세포침습실험결과현시,TGF-β1조천과섬유막적세포수여공백조、TGF-β1+ NSC23766조、NSC23766조천과섬유막적세포수비교,차이무통계학의의(F=0.371,P=0.055).응효수축실험결과현시,TGF-β1조능명현증가세포적촉응효수축능력,조간차이유통계학의의(F=40.473,P=0.014),TGF-β1조상대TGF-β1+ NSC23766조,차이유통계학의의(P=0.022).결론 Rac1재TGF-β1유도RPE세포응효수축개변중발휘작용;NSC23766가통과조공Rac1활화억제RPE세포학행위적개변.
Objective To study the role of Racl in the epithelial-mesenchymal transition (EMT) process of retinal pigment epithelial cells (RPE) induced by transforming growth factor β (TGF-β).Methods Human ARPE-19 cells were divided into 4 groups including control group,TGF-β group,TGF-β + NSC23766 group,NSC23766 group.NSC23766 was added to medium 2 hours before TGF-β treatment to block the Rac1 receptors.α-smooth muscle actin (α-SMA) expression was measured by immunofluorescence and Western blot.Cell scratch assay,invasion assay and gel contraction experiments were used to measure cell migration,invasion,cell contraction.Results The expression of α-SMA was higher in TGF-β group,compared with the control group,TGF-β + NSC23766 group (F=825.314,P＜0.05).Cell scratch assay showed that the cellular gap was less in GF-β group,compared with the control group,TGF-β + NSC23766 group,NSC23766 group (F=177.351,P＜0.05).Cell invasion assay showed that,the number of cells pass through the fiber membrane was the same in TGF-β group and other 3 groups (F=0.371,P=0.055).Gel contraction assay showed that TGF-β can promote the cellular contraction,compare to the control group,TGF-β + NSC23766 group,NSC23766 group,the difference was statistically significant (F=40.473,P＜0.05).Conclusion Rac1 play a role in TGF-β-induced behavioral changes of RPE cells; NSC23766 inhibit RPE cellular behavior change by regulating Rac1 activation.
