CAJ | 학술논문

目的通过研究经shRNA慢病毒转染串珠素的大鼠硬膜外瘢痕组织成纤维细胞的增殖能力,探讨其对硬膜外瘢痕组织形成的影像. 方法 30只Wistar大鼠行椎板切除术,建立模型,于培养4周后提取硬膜外瘢痕组织.通过组织块贴壁法培养硬膜外瘢痕组织中的成纤维细胞并将细胞分为3组:未转染组(空白对照组)、荧光蛋白(GFP)慢病毒转染组(GFP组)和shRNA慢病毒转染组(shRNA组,内含GFP).成纤维细胞转染成功后,分别通过Western-blot、荧光定量PCR及MTT法检测3组的成纤维细胞的RNA、蛋白质表达及细胞增殖的差异. 结果 Western-blot检测显示shRNA组串珠素蛋白明显低于空白对照组和GFP组.荧光定量PCR检测显示shRNA组串珠素RNA表达低于空白对照组和GFP组.MTT检测显示0h各组吸光度(A)值差异均无统计学意义(P＞0.05),24 ～ 96 hshRNA组吸光度值(A)低于空白对照组和GFP组,差异均有统计学意义(P＜0.05). 结论经shRNA慢病毒转染串珠素能抑制大鼠硬膜外瘢痕组织成纤维细胞的增殖.
목적 통과연구경shRNA만병독전염천주소적대서경막외반흔조직성섬유세포적증식능력,탐토기대경막외반흔조직형성적영상. 방법 30지Wistar대서행추판절제술,건립모형,우배양4주후제취경막외반흔조직.통과조직괴첩벽법배양경막외반흔조직중적성섬유세포병장세포분위3조:미전염조(공백대조조)、형광단백(GFP)만병독전염조(GFP조)화shRNA만병독전염조(shRNA조,내함GFP).성섬유세포전염성공후,분별통과Western-blot、형광정량PCR급MTT법검측3조적성섬유세포적RNA、단백질표체급세포증식적차이. 결과 Western-blot검측현시shRNA조천주소단백명현저우공백대조조화GFP조.형광정량PCR검측현시shRNA조천주소RNA표체저우공백대조조화GFP조.MTT검측현시0h각조흡광도(A)치차이균무통계학의의(P＞0.05),24 ～ 96 hshRNA조흡광도치(A)저우공백대조조화GFP조,차이균유통계학의의(P＜0.05). 결론 경shRNA만병독전염천주소능억제대서경막외반흔조직성섬유세포적증식.
Objective To research the proliferation of fibroblasts in epidural scar transfected by perlecan shRNA lentiviral particles.Methods Experimental models were established in 30 Wistar mice which underwent laminectomy.The epidural scar tissue was harvested after culture for 4 days.The fibroblasts in the epidural scar tissue were cultured by tissue explant adherence before they were randomly assigned into no transfection (control group),green fluorescent protein (GFP) lentiviral transfection (GFP group) and shRNA lentiviral transfection (shRNA group).After the fibroblasts were successfully transfected,their expressions of perlecan mRNA and protein and proliferation were detected by Western-blot,RT-PCR and MTT and compared as well among the 3 groups.Results Western-blot and RT-PCR showed the expressions of perlecan mRNA and protein were significantly lower in the shRNA group than in the control and GFP groups (P ＜ 0.05).MTT showed no significant difference among the 3 groups in the optical density at 0 hour (P ＞0.05) but significantly lower optical density in the shRNA group than in the control and GFP groups at 24 to 96 hours (P ＜ 0.05).Conclusion Growth of fibroblasts in epidural scar can be inhibited significantly by transfection with perlecan shRNA lentiviral particles.