肝衰竭环境下清热解毒凉血化瘀法对骨髓间充质干细胞影响的体外实验研究中西医结合临床专业论文.docxVIP

学差异。但在细胞培养10天后观察发现，中药干预组细胞生长明显 学差异。但在细胞培养10天后观察发现，中药干预组细胞生长明显 优于肝衰竭组。 2、BMSCs制作细胞爬片，培养10天后进行CKl8免疫荧光检测。 结果显示健康组染色为阴性，肝衰竭组及中药干预组染色为阳性，后 者染色较强，但均明显弱于肝细胞染色。推测BMSCs肝向分化过程中 细胞分化数量或细胞成熟度较低。 3、BMSCs培养10天后通过Western blot法检测相关蛋白表达。 ALB蛋白表达结果显示：健康组、肝衰竭组、中药干预组均有表达， 且依次递增，各组之间有统计学差异，但其表达量较少。AFP蛋白表 达结果显示：健康组、肝衰竭组、中药干预组表达强度依次增强，各 组间差异有统计学意义。 4、BMSCs培养10天后通过Western blot法检测IDO表达，结 果显示：健康组IDO蛋白无明显表达。IDO蛋白在中药干预组中的表 达高于肝衰竭组，且两组间差异具有统计学意义。 研究结论： 通过建立BMSCs移植治疗肝衰竭体外模型，观察发现BMSCs在肝 衰竭环境中增殖活性相对下降，具有肝向分化趋势，及免疫调节活性。 清热解毒凉血化瘀中药在肝衰竭环境中对BMSCs的增殖在短时间内 无明显作用，但可以促进干细胞分化，且明显提高干细胞免疫调节活 性。 关键词： 中药；骨髓间充质干细胞；肝衰竭；免疫调节 万方数据 In In vitro experiment study on the effect of Clearing—heat and Detoxicating Combined with Cooling Blood and Removing Blood Stasis on bone marrow mesenchymal stem cells in the environment of hepatic failure Abstract Objective：Hepatic failure simulation environment was established by the rat serum，and the mesenchymal stem cells were cultured in vitro．To observe the mesenchymal stem cell proliferation，differentiation and immune regulation in liver failure environment，and the effect of Chinese medicine on mesenchymal stem cells in the environment of hepfatic failure．To further explore the detoxification method of cooling blood to remove stasis on liver failure in bone marrow mesench)rmal stem cell and synergistic effect at the cellular level． Method： 1．The screening of 1 rat BMSCs were isolated and cultured and passaged．Extraction，separation，from 3-4 week old SPF SD rats bone marrow culture BMSCs．Using the whole bone marrow culture method and passaged screening method for cell culture and purification．When the cell proliferation at 80～90％were passaged．For the detection and further transplant cells to the third generation．Through flow cytometry identified cells were used for BMSCs． ．4． 万方数据 2．the 2．the experimental serum preparation：30 rats were divided into model group and healthy group，2 1 rats in the model group model of liver failure made by intraperitoneal inj ection of D-GaIN．Aft