中华普通外科杂志
中華普通外科雜誌
중화보통외과잡지
CHINESE JOURNAL OF GENERAL SURGERY
2015年
4期
304-307
,共4页
俞富祥%黄立栋%唐银河%古妮%张启瑜
俞富祥%黃立棟%唐銀河%古妮%張啟瑜
유부상%황립동%당은하%고니%장계유
间质干细胞%胰腺%细胞增殖%细胞凋亡%细胞因子类
間質榦細胞%胰腺%細胞增殖%細胞凋亡%細胞因子類
간질간세포%이선%세포증식%세포조망%세포인자류
Mesenchymal stem cells%Pancreas%Cell proliferation%Apoptosis%Cytokines
目的 探讨改良大鼠胰腺星状细胞(pancreatic stellate cells,PSCs)的分离提取,同时探讨脂肪间质干细胞(adipose derived stem cells,ADSCs)对活化态PSCs增殖、活化、凋亡的影响.方法 采用不灌注,直接提取胰腺剪碎消化,Optiprep密度梯度离心法分离大鼠PSCs.建立PSCs与ADSCs的双层培养体系,CCK-8比色法检测ADSCs对PSCs增殖的影响;Western blot检测PSCs α-肌动蛋白(αt-SMA)的表达;流式细胞仪检测ADSCs对PSCs凋亡的影响;检测培养液中细胞因子的含量探讨可能的作用机制.结果 PSCs得率稳定在5×106/鼠以上,纯度为90％ ～95％左右,存活率为92％ ～97％.ADSCs明显抑制PSCs的活化与增殖(F=223.27,P＜0.05;F=52.97,P＜0.05),促进PSCs的凋亡(F =43.62,P＜0.05),ADSCs比PSCs分泌更多的NGF(14.68±0.94比8.31±0.86,t=4.67,P＜0.05),而较少分泌TGF-β1(10.65±0.46比70.47±0.99,t=21.72,均P＜0.01).结论 大鼠ADSCs具有分泌细胞因子抑制PSCs活化增殖,促进凋亡的潜能.
目的 探討改良大鼠胰腺星狀細胞(pancreatic stellate cells,PSCs)的分離提取,同時探討脂肪間質榦細胞(adipose derived stem cells,ADSCs)對活化態PSCs增殖、活化、凋亡的影響.方法 採用不灌註,直接提取胰腺剪碎消化,Optiprep密度梯度離心法分離大鼠PSCs.建立PSCs與ADSCs的雙層培養體繫,CCK-8比色法檢測ADSCs對PSCs增殖的影響;Western blot檢測PSCs α-肌動蛋白(αt-SMA)的錶達;流式細胞儀檢測ADSCs對PSCs凋亡的影響;檢測培養液中細胞因子的含量探討可能的作用機製.結果 PSCs得率穩定在5×106/鼠以上,純度為90％ ～95％左右,存活率為92％ ～97％.ADSCs明顯抑製PSCs的活化與增殖(F=223.27,P＜0.05;F=52.97,P＜0.05),促進PSCs的凋亡(F =43.62,P＜0.05),ADSCs比PSCs分泌更多的NGF(14.68±0.94比8.31±0.86,t=4.67,P＜0.05),而較少分泌TGF-β1(10.65±0.46比70.47±0.99,t=21.72,均P＜0.01).結論 大鼠ADSCs具有分泌細胞因子抑製PSCs活化增殖,促進凋亡的潛能.
목적 탐토개량대서이선성상세포(pancreatic stellate cells,PSCs)적분리제취,동시탐토지방간질간세포(adipose derived stem cells,ADSCs)대활화태PSCs증식、활화、조망적영향.방법 채용불관주,직접제취이선전쇄소화,Optiprep밀도제도리심법분리대서PSCs.건립PSCs여ADSCs적쌍층배양체계,CCK-8비색법검측ADSCs대PSCs증식적영향;Western blot검측PSCs α-기동단백(αt-SMA)적표체;류식세포의검측ADSCs대PSCs조망적영향;검측배양액중세포인자적함량탐토가능적작용궤제.결과 PSCs득솔은정재5×106/서이상,순도위90％ ～95％좌우,존활솔위92％ ～97％.ADSCs명현억제PSCs적활화여증식(F=223.27,P＜0.05;F=52.97,P＜0.05),촉진PSCs적조망(F =43.62,P＜0.05),ADSCs비PSCs분비경다적NGF(14.68±0.94비8.31±0.86,t=4.67,P＜0.05),이교소분비TGF-β1(10.65±0.46비70.47±0.99,t=21.72,균P＜0.01).결론 대서ADSCs구유분비세포인자억제PSCs활화증식,촉진조망적잠능.
Objective To evaluate a new method for the isolation of rat pancreatic stellate cells (PSCs) and to investigate the influence of adipose derived stem cells (ADSCs) on PSCs in vitro.Methods Normal rat PSCs was isolated by collagenase and Optiprep density gradient centrifugation.The coculture system of ADSCs and PSCs was set up by transwell insert.The proliferation of PSCs was tested by CCK-8 test kit.Smoothmuscle α-actin (α-SMA) expression of PSCs were tested by Western blot.The apoptosis of HSCs was tested by flow cytometer.The cytokines in the culture solution were tested by ELISA kit.Results The quantity of PSCs was above 5 × 106 cells per rat.The purity and the viability of the cells were about 90-97 percent.After coculture for 72 h,the proliferation and activation of PSCs was inhibited by ADSCs (F =223.27,P ＜ 0.05 ; F =52.97,P ＜ 0.05) and the apoptosis of PSCs was promoted by ADSCs (F =43.62,P ＜ 0.05).more NGF and less TGF-β1 was secreted by ADSCs than by PSCs (NGF:14.68 ± 0.94 vs.8.31 ±0.86,t =4.67,P ＜0.05;TGF-β1:10.65 ±0.46 vs.70.47 ±0.99,t =21.72,P＜0.01).Conclusions ADSCs inhibit the proliferation and activation of PSCs by ADSCs secreting cytokines.
