吴雄健++郑虹+朱海燕+毛忠懿+张蕾++谢军

[摘要]目的 探討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。

[关键词]骨髓间质干细胞移植；肝星状细胞；凋亡情况；肝纤维化；治疗机制

[中图分类号] R575.5 [文献标识码] A [文章编号] 1674-4721（2017）07（c）-0107-04

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

WU Xiong-jian1 ZHENG Hong2 ZHU Hai-yan1 MAO Zhong-yi1 ZHANG Lei1 XIE Jun1

1.Department of Gastroenterology，the First Affiliated Hospital of Gannan Medical University，Jiangxi Province，Ganzhou 341000，China；2.Gannan Medical University，Jiangxi Province，Ganzhou 341000，China

[Abstract]Objective To investigate the apoptosis of hepatic stellate cells 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 Center，Zhongshan 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 cells 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=10），which was not performed with intervention.The purity identification of bone marrow 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 CCl4，the content of hydroxyproline in the rat′s liver in the experimental group was significantly higher than that before transplantation （P<0.05）.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）.Conclusion The hepatic stellate cell apoptosis induced by bone marrow mesenchymal stem cells may be a major mechanism for the treatment of hepatic fibrosis.

[Key words]Bone marrow mesenchymal stem cell transplantation；Hepatic stellate cells；Apoptosis；Hepatic fibrosis；Treatment mechanism

肝纤维化属于一种病理表现，各种慢性肝病向一定阶段发展后均会表现出来，现阶段，临床还没有特异性的方法治疗肝纤维化及其终末阶段的肝硬化。在肝纤维化发生及缓解中，肝星状细胞激活及凋亡异常占有极为重要的地位。近年来，相关医学研究显示，在肝纤维化或肝硬化的治疗中，骨髓间质干细胞能够将一定的作用发挥出来，但是现阶段临床还未弄清楚其具体机制[1-2]。为了将有效的理论依据进一步提供给骨髓间质肝细胞对肝纤维化的治疗，本研究探讨了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，现报道如下。……