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目的 :探索调肝益气通络方对大鼠骨髓间充质干细胞(MSC)增殖能力的影响。方法 :腹腔注射“CCl4-橄榄油”混悬液制备WISTAR大鼠(35只)肝纤维化模型;其中30只灌胃给予调肝益气通络方(低、中、高剂量)1个月,制备含药血清;另外5只用于MSC细胞的分离与培养;用含药血清诱导MSC向肝细胞的分化连续14 d;在各检测点进行形态学观察通过MTT对细胞增殖情况进行评价,通过Western Blot检测诱导第7天、第14天甲胎蛋白的表达程度。结果 :各浓度含药血清在加入1~14 d后,其细胞数量均比常规培养组增多,并同用药剂量、时间呈正比;诱导14 d后高剂量组甲胎蛋白呈强阳性表达。结论 :调肝益气通络方对大鼠骨髓间充质干细胞的增殖能力具有显著的促进作用,并可促进甲胎蛋白的分泌。
Objective: To explore the effects of Tiaogan Yiqi Tongluo Recipe on the proliferation of rat bone marrow mesenchymal stem cells (MSCs). Methods: Wistar rats (35 rats) were injected intraperitoneally with “CCl4-olive oil” suspension to establish liver fibrosis model. 30 of them were fed with Tiaogan Yiqi Tongluo Recipe (low, middle and high dose) 1 And the other 5 were used for isolation and culture of MSC cells; MSCs were induced to differentiate into hepatocytes for 14 days continuously with drug-containing serum; Morphological observation was performed at each detection point. Cell proliferation was performed by MTT The level of alpha-fetoprotein expression on day 7 and 14 was detected by Western Blot. Results: After 1 to 14 days of treatment, the number of cells in each group increased more than that in routine culture group, and was proportional to the dose and time. After 14 days of induction, the high-dose group showed a strong positive expression of alpha-fetoprotein. Conclusion: Tiaogan Yiqi Tongluo Recipe can significantly promote the proliferation of rat bone marrow mesenchymal stem cells and promote the secretion of alpha-fetoprotein.