摘要
目的观察左归丸及其拆方的粗提取物对骨髓干细胞转化肝细胞的影响,为进一步探索左归丸的有效成分奠定实验基础。方法利用透过性支持物(聚酯TranswellR透明嵌套)建立骨髓干细胞-肝组织细胞共培养体系,采用水煮醇沉方法提取制备左归丸及其拆方粗提取物加入共培养体系。骨髓干细胞与肝细胞共培养7、14、21、28、35d时分别取透过性支持物中的骨髓干细胞爬片用PAS法检测糖原,免疫细胞化学方法(ICC法)检测肝细胞标志物(AFP、CK18、Alb)。结果左归丸及其拆方粗提取物浓度高于1.56 mg/ml时,影响骨髓干细胞的存活,而浓度等于1.56 mg/ml时,共培养条件下骨髓干细胞的糖原、AFP、Alb、CK18的表达量,平行对照各组差异无统计学意义(P>0.05)。结论骨髓干细胞对培养条件比较苛刻,一般的中药粗提制剂不适于骨髓干细胞-肝细胞共培养体系的实验。
Objective To observe the influence of Zuogui Wan extract on bone marrow stem cell transforming liver cell,and establish experimental basis to further study effective constituent of Zuogui Wan.Methods TranswellR insert was used to establish bone marrow stem cell-liver cell co-culture system.Zuogui Wan extract was prepared by water boiled-alcohol sink method,and was added to co-culture system.Bone marrow stem cell in TranswellR insert was detected after co-culture 7,14,21,28 and 35 days.Glycogen was detected by PAS method,liver cell markers（AFP,Alb and CK18） were detected by immunocytochemistry method.Results Zuogui Wan extract influenced bone marrow stem cell survive when its concentration higher than 1.56mg/ml.The expression of glycogen,AFP,Alb and CK18 in bone marrow stem cell had no significant difference contrasted to control groups,when Zuogui Wan extract＇s concentration was 1.56mg/ml in co-culture system.Conclusion Bone marrow stem cell was sensitive to culture condition.Traditional Chinese medicine extract prepared by water boiled-alcohol sink method was unfit for experiment of bone marrow stem cell-liver cell co-culture system.
出处
《临床肝胆病杂志》
CAS
2012年第3期192-195,200,共5页
Journal of Clinical Hepatology
基金
国家自然科学基金(90709041
30672590
30271562
30371787
81102531)
关键词
左归丸
骨髓祖代细胞
肝细胞
ZUOGUI PILL
myeloid progenitor cells
hepatocytes
