中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
5期
1116-1118
,共3页
谢华%农鲁明%高共鸣%周栋%黄永静
謝華%農魯明%高共鳴%週棟%黃永靜
사화%농로명%고공명%주동%황영정
G蛋白耦联受体激酶结合蛋白1%周期性应力%髓核细胞
G蛋白耦聯受體激酶結閤蛋白1%週期性應力%髓覈細胞
G단백우련수체격매결합단백1%주기성응력%수핵세포
G protein coupled receptor kinase interacting protein 1%Periodic mechanical stress%Nucleus pulposus cell
目的 观察周期性应力下G蛋白耦联受体激酶结合蛋白1(Git1)对大鼠髓核细胞细胞外基质表达的影响.方法 首先将细胞玻片分为对照组和加压组,通过Western blot检测两组Git1蛋白的表达,荧光实时定量聚合酶链反应(FQ-PCR)检测两组细胞外基质的表达变化.然后使用小干扰RNA(siRNA)阻断Git1蛋白,分为对照组、Git1 siRNA组、siRNA阴性对照组,在加压环境下比较各组细胞外基质的表达变化.结果 加压组细胞的Git1蛋白表达(1.372±0.205)较对照组(0.478 ±0.186)明显增高,加压组细胞外基质的基因表达较对照组明显升高,差异有统计学意义(P＜0.05).siRNA处理细胞后,Git1 siRNA组Git1蛋白表达(0.587±0.142)较对照组(1.041±0.103)明显降低.加压环境下,Git1 siRNA组细胞外基质的基因表达较对照组和siRNA阴性对照组明显下降,差异有统计学意义(P＜0.05).结论 周期性压力能促进髓核细胞细胞外基质的表达,Git1在压力传导中起信号传递作用.
目的 觀察週期性應力下G蛋白耦聯受體激酶結閤蛋白1(Git1)對大鼠髓覈細胞細胞外基質錶達的影響.方法 首先將細胞玻片分為對照組和加壓組,通過Western blot檢測兩組Git1蛋白的錶達,熒光實時定量聚閤酶鏈反應(FQ-PCR)檢測兩組細胞外基質的錶達變化.然後使用小榦擾RNA(siRNA)阻斷Git1蛋白,分為對照組、Git1 siRNA組、siRNA陰性對照組,在加壓環境下比較各組細胞外基質的錶達變化.結果 加壓組細胞的Git1蛋白錶達(1.372±0.205)較對照組(0.478 ±0.186)明顯增高,加壓組細胞外基質的基因錶達較對照組明顯升高,差異有統計學意義(P＜0.05).siRNA處理細胞後,Git1 siRNA組Git1蛋白錶達(0.587±0.142)較對照組(1.041±0.103)明顯降低.加壓環境下,Git1 siRNA組細胞外基質的基因錶達較對照組和siRNA陰性對照組明顯下降,差異有統計學意義(P＜0.05).結論 週期性壓力能促進髓覈細胞細胞外基質的錶達,Git1在壓力傳導中起信號傳遞作用.
목적 관찰주기성응력하G단백우련수체격매결합단백1(Git1)대대서수핵세포세포외기질표체적영향.방법 수선장세포파편분위대조조화가압조,통과Western blot검측량조Git1단백적표체,형광실시정량취합매련반응(FQ-PCR)검측량조세포외기질적표체변화.연후사용소간우RNA(siRNA)조단Git1단백,분위대조조、Git1 siRNA조、siRNA음성대조조,재가압배경하비교각조세포외기질적표체변화.결과 가압조세포적Git1단백표체(1.372±0.205)교대조조(0.478 ±0.186)명현증고,가압조세포외기질적기인표체교대조조명현승고,차이유통계학의의(P＜0.05).siRNA처리세포후,Git1 siRNA조Git1단백표체(0.587±0.142)교대조조(1.041±0.103)명현강저.가압배경하,Git1 siRNA조세포외기질적기인표체교대조조화siRNA음성대조조명현하강,차이유통계학의의(P＜0.05).결론 주기성압력능촉진수핵세포세포외기질적표체,Git1재압력전도중기신호전체작용.
Objective To evaluate the regulation effect of the G protein coupled receptor kinase interacting protein 1 (Git1) on extracellular matrix of nucleus pulplsus cells under periodic mechanical stress.Methods There were two parts in the experiment.In the first part,some cell slides were randomized into two groups:the stress group and the control group.The Git1 protein expression was detected by Western blotting and the variation of extracellular matrix was evaluated by real-time fluorescent quantitative polymerase chain reaction (FQ-PCR).In the second part,some cell slides were randomized into three groups:control group,Git1 small interfering RNA (siRNA) group and siRNA negative control group,in which the changes of extracellular matrix were tested by FQ-PCR in the stress situation.Results Expression of Git1 protein in the stress group (1.372 ±0.205) was significantly higher than that in the control group (0.478 ± 0.186).The level of extracellular matrix in the stress group was much higher in the control group.Git1 protein expression was dramatically decreased in Git1 siRNA group (0.587 ± 0.142) compared to control group (1.041 ± 0.103).In stress situation,the gene expression of extracellular matrix was significantly decreased in Git1 siRNA group compared with control group and siRNA negative control group.Conclusion Periodic mechanical stress can promote the gene expression of extracellularmatrix of rat nucleus pulposus cells.Git1 protein plays important role in the mechanical transductions.
