骨髓间质干细胞移植后肝星状细胞凋亡情况及其治疗肝纤维化的机制探讨

Exploration of the apoptosis of hepatic stellate cells after bone marrow mesenchymal stem cell transplantation and its mechanism in the treatment of hepatic fibrosis

目的探讨骨髓间质干细胞移植后肝星状细胞凋亡情况及其治疗肝纤维化的机制。方法购买中山大学动物实验中心提供的30只清洁级SD大鼠,实验研究选取20只,将提取骨髓间质干细胞的10只作为实验组,建立肝纤维化大鼠模型,骨髓干细胞动员干预大鼠肝纤维化及肝星状细胞凋亡,检测肝纤维化及星状细胞凋亡指标;将未提取骨髓间质干细胞的10只作为对照组,不对其进行干预。然后对骨髓间质干细胞的纯度鉴定结果进行分析,并对两组大鼠的肝脏羟脯氨酸含量、肝星状细胞激活及凋亡情况进行统计分析。结果 CD34、CD44、CD45、CD95阴性细胞分别为99.75%、99.51%、99.80%、95.72%。实验组大鼠CCl4皮下注射诱导8周后的肝脏羟脯氨酸含量显著高于骨髓间质干细胞移植前(P<0.05);实验组大鼠骨髓间质干细胞移植1周后的肝脏羟脯氨酸含量显著低于骨髓间质干细胞移植3 d后(P<0.05),对照组大鼠骨髓间质干细胞移植1周后的肝脏羟脯氨酸含量显著高于骨髓间质干细胞移植3 d后(P<0.05);实验组大鼠骨髓间质干细胞移植1周后的肝脏羟脯氨酸含量显著低于对照组(P<0.05)。实验组大鼠骨髓间质干细胞移植1周后的肝星状阳性细胞数显著少于骨髓间质干细胞移植3 d后(P<0.05);实验组大鼠骨髓间质干细胞移植1周后的肝星状阳性细胞数显著少于对照组(P<0.05)。实验组大鼠骨髓间质干细胞移植3 d后的肝星状凋亡细胞数显著多于骨髓间质干细胞移植前及对照组(P<0.05)。结论骨髓间质干细胞移植后诱导肝星状细胞凋亡可能是其治疗肝纤维化的一个主要机制。

Objective To investigate the apoptosis of hepatic stellate ceils after bone marrow mesenchymal stem cell transplantation and its mechanism in the treatment of hepatic fibrosis.Methods 30 clean SD rats were purchased from Animal Experimental EenteruZhongshan University,and 20 of them were prepared for experimental research.Only ten rats whose bone marrow mesenchym stem cells were extracted and selected as the experimental group for the establishment of rat model with hepatic fibrosis.The hepatic fibrosis and apoptosis of hepatic stellate ceils were intervened by bone marrow stem cell mobilization to test these indices.Another group without extraction of bone marrow mesenchymal stem cells was classified into the control group （n=lO）,which was not performed with intervention.The purity identification of bone mar- row mesenchymal stem cells was analyzed,and the hydroxyproline content as well as hepatic stellate cell activation and apoptosis Were statistically analyzed.Results The percentage of CD34,CD44,CD45 and CD95 in negative cells were 99.75%,99.51%,99.80% and 95.72%,respectively.After 8-week induction via subcutaneous injection of CC14,the content of hydroxyproline in the rat＇s liver in the experimental group was significantly higher than that before transplantation （P〈O.O5）.In the experimental group,the content of hydroxyproline in the liver after one-week bone marrow mesenchymal stem cell transplantation was much lower compared with that 3-day after transplantation （P〈0.05）.In the control group,the content of hydroxyproline in the liver after one.week bone marrow mesenchymal stem cell transplantation was greatly higher in comparison with that 3-day after transplantation （P〈0.05）.The content of hydroxyproline in the liver after one-week bone marrow mesenchymal stem cell transplantation in the experimental group was remarkably lower than that in the control group （P〈0.05）.The amount of hepatic stellate positive cells in the experimental group one-week after bone marrow mesenchymal stem cell transplantation was greatly less than that in the control group （P〈0.05）.The amount of hepatic stellate apoptotic cells 3-day after bone marrow mesenchymal stem cell transplantation in the experimental group was remarkably more than that before transplantation and in the control group （P〈0.05）.Conelusion The hepatic stellate cell apoptosis induced by bone marrow mesenehymal stem cells may be a major mechanism for the treatment of hepatic fibrosis.