脂肪间质干细胞抑制大鼠胰腺星状细胞活性的实验研究  

The effect of adipose derived stem cells on pancreatic stellate cells in vitro

作　　者：俞富祥[1] 黄立栋[1] 唐银河[1] 古妮 张启瑜[1] 

机构地区：[1]温州医科大学附属第一医院肝胆胰外科,浙江省325000

出　　处：《中华普通外科杂志》2015年第4期304-307,共4页Chinese Journal of General Surgery

基　　金：浙江省重中之重外科学组基金资助项目（浙教高科2008-255）;浙江省教育厅科研项目基金资助项目（Y201326678）

摘　　要：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；Westernblot检测PSCs α-肌动蛋白（α-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5×10^6／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。Objective To evaluate a new method for the isolation of rat pancreatic stellate cells （PSCs） and to investigate the influence of adipose derived stem ceils （ADSCs） on PSCs in vitro. Methods Normal rat PSCs was isolated by collagenase and Optiprep density gradient centrifugation. The cocuhure 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 （ct-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 × 10^6 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-131 was secreted by ADSCs than by PSCs（NGF：14.68 ：e0.94 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.

关 键 词：间质干细胞 胰腺 细胞增殖 细胞凋亡 细胞因子类 

分 类 号：R322.4[医药卫生—人体解剖和组织胚胎学]