CAJ | 학술논문

目的：观察沉默Smo基因后对大鼠原代软骨细胞增殖与凋亡的影响。方法用机械-酶消化法获取原代大鼠软骨细胞，经免疫细胞化学Ⅱ型胶原（ ColⅡ）鉴定后，将实验分为对照组、control siRNA组和Smo siRNA 1～3组，以慢病毒为载体将siRNA转入软骨细胞，72 h后，MTT法检测细胞活力；反转录PCR （ RT-PCR）及蛋白印迹（ Western blot）检测Smo的表达量；流式细胞术检测细胞凋亡率。结果各组序列经慢病毒载体成功转染到原代软骨细胞中， Smo siRNA1～3均不同程度的抑制Smo的表达，以Smo siRNA2组最为明显，其mRNA及蛋白的表达分别为0.19±0.03和0.39±0.07；同时，Smo siRNA2组细胞活力最低（77.38％±7.19％）而凋亡率最高（21.43％±2.97％）。结论沉默Smo可抑制原代软骨细胞增殖，并诱导软骨细胞凋亡，Smo具有保护软骨细胞免遭凋亡的作用。
목적：관찰침묵Smo기인후대대서원대연골세포증식여조망적영향。방법용궤계-매소화법획취원대대서연골세포，경면역세포화학Ⅱ형효원（ ColⅡ）감정후，장실험분위대조조、control siRNA조화Smo siRNA 1～3조，이만병독위재체장siRNA전입연골세포，72 h후，MTT법검측세포활력；반전록PCR （ RT-PCR）급단백인적（ Western blot）검측Smo적표체량；류식세포술검측세포조망솔。결과각조서렬경만병독재체성공전염도원대연골세포중， Smo siRNA1～3균불동정도적억제Smo적표체，이Smo siRNA2조최위명현，기mRNA급단백적표체분별위0.19±0.03화0.39±0.07；동시，Smo siRNA2조세포활력최저（77.38％±7.19％）이조망솔최고（21.43％±2.97％）。결론침묵Smo가억제원대연골세포증식，병유도연골세포조망，Smo구유보호연골세포면조조망적작용。
Objective To investigate the effects of silencing Smo gene on proliferation and apoptosis of rat prima-ry chondrocyte in vitro.Methods The primary chondrocyte was obtained by mechanical-enzyme digestion and identified by Immunohistochemical cells ( ColⅡ) .The animals were divided into control group , control siRNA group and Smo siRNA 1 ~3 group.The siRNA was transfected into chondrocytes by lentivirus vector .After 72 h, the cell viability was detected by MTT, Smo expression was detected by RT-PCR and Western blot, and the apoptosis of chondrocyte was assessed by flow cytometry .Results All types of siRNA were transfected into primary chondrocyte by vectors, the Smo siRNA 1 ~3 may inhibit the expression of Smo mRNA and protein in chondrocytes, and Smo siRNA2 had the highest silencing rate ( the expressions of Smo mRNA and protein were 0.19 ±0.03 and 0.39 ±0.07 ) .The cell viability in Smo siRNA2 group was lowest ( 77.38% ±7.19%) , while the apoptosis rate of Smo siRNA2 was highest ( 21.43%±2.97%) .Conclusions Silencing Smo gene in primary chondrocytes may inhibit proliferation and promote apoptosis , Smo may have a protecting role from apop-tosis of the chondrocyte.