[氧化苦参碱可促进肝纤维化大鼠间充质干细胞治疗：一项实验研究]

Chai Ning-Li, Xu Shi-Ping, Wan Jun, Wu Ben-Yan

Department of Gastroenterology, Division of South Building, Chinese People's Liberation Army General Hospital, Beijing 100853, China.

Zhongguo Zhong Xi Yi Jie He Za Zhi. 2013 Jun;33(6):840-4.

OBJECTIVE

To investigate whether oxymatrine (OM) could promote mesenchymal stem cell (MSC) therapy in CCl4-induced hepatic fibrosis (HF) in rats and to initially explore its mechanisms.

METHODS

Totally 50 male SD rats were randomly divided into five groups,i.e., the normal control group, the model group, the MSC therapy group, the OM therapy group, and the MSC combined OM therapy group, 10 in each group. Except the normal control group, the HF model was duplicated by CCl4 induction. After successful modeling, rats in the MSC therapy group received 5 x10(6) MSCs by intravenous injection via caudal vein, once a week. Rats in the OM therapy group received 50 mg/kg OM by intramuscular injection, three times a week. Rats in MSC combined OM therapy group received 5 x 10(6) MSCs by intravenous injection via caudal vein, once a week and 50 mg/kg OM by intramuscular injection three times a week. Equal volume of normal saline was given to those in the normal control group and the model group. All medication lasted for 8 weeks. Serum levels of ALT and AST were detected 8 weeks later. The hepatic histopathological injury and extracellular matrix deposit were assessed using HE and Masson staining. Expressions of serum interleukin-4 (IL-4) and interleukin-10 (IL-10) were detected using enzyme linked immunosorbent assay (ELISA).

RESULTS

(1) Compared with the normal control group, serum levels of ALT and AST significantly increased in the model group (P < 0.05). Compared with the model group, serum levels of ALT and AST significantly decreased in the OM therapy group, the MSC therapy group, and the MSC combined OM therapy group at the end of 8 weeks of treatment (P < 0.05). But serum levels of ALT and AST were significantly lower in the MSC combined OM therapy group than in the OM therapy group and the MSC therapy group (P < 0.05). (2) Compared with the model group, the hepatic injury was significantly lessened and the area of extracellular matrix deposit was significantly reduced in the OM therapy group, the MSC therapy group, and the MSC combined OM therapy group (P < 0.05). Besides, they wer more significant in the MSC combined OM therapy group (P < 0.05). (3) Compared with the model group, the serum IL-4 level was significantly higher in the MSC therapy group and the MSC combined MO group (P < 0.05). It was higher in the MSC combined MO group (P < 0.05). Although the serum IL-4 level also increased in the OM therapy group, but with no statistical difference (P > 0.05). (4) The serum IL-10 level significantly increased in the OM therapy group, the MSC therapy group, and the MSC combined OM therapy group (P < 0.05), and it was the highest in the MSC combined OM therapy group among the three groups (P < 0.05). (5) Two-photon fluorescence imaging showed no signals of MSCs in liver with or without OM injection.

CONCLUSION

OM could promote mesenchymal stem cell therapy in hepatic fibrosis rats, which might be involved in increasing serum levels of IL-4 and IL-10.

目的

探讨氧化苦参碱（OM）能否促进间充质干细胞（MSC）对大鼠四氯化碳诱导的肝纤维化（HF）的治疗作用，并初步探讨其机制。

方法

将50只雄性SD大鼠随机分为5组，即正常对照组、模型组、MSC治疗组、OM治疗组和MSC联合OM治疗组，每组10只。除正常对照组外，其余组采用四氯化碳诱导建立肝纤维化模型。造模成功后，MSC治疗组大鼠通过尾静脉每周1次静脉注射5×10⁶个MSC；OM治疗组大鼠每周3次肌肉注射50mg/kg OM；MSC联合OM治疗组大鼠通过尾静脉每周1次静脉注射5×10⁶个MSC，同时每周3次肌肉注射50mg/kg OM。正常对照组和模型组大鼠给予等体积生理盐水。所有用药持续8周。8周后检测血清谷丙转氨酶（ALT）和谷草转氨酶（AST）水平。采用苏木精-伊红（HE）和Masson染色评估肝脏组织病理学损伤及细胞外基质沉积情况。采用酶联免疫吸附测定（ELISA）法检测血清白细胞介素-4（IL-4）和白细胞介素-10（IL-10）的表达。

结果

（1）与正常对照组相比，模型组血清ALT和AST水平显著升高（P<0.05）。治疗8周结束时，与模型组相比，OM治疗组、MSC治疗组和MSC联合OM治疗组血清ALT和AST水平显著降低（P<0.05）。但MSC联合OM治疗组血清ALT和AST水平显著低于OM治疗组和MSC治疗组（P<0.05）。（2）与模型组相比，OM治疗组、MSC治疗组和MSC联合OM治疗组肝损伤明显减轻，细胞外基质沉积面积显著减少（P<0.05）。此外，MSC联合OM治疗组上述变化更显著（P<0.05）。（3）与模型组相比，MSC治疗组和MSC联合OM治疗组血清IL-4水平显著升高（P<0.05）。MSC联合OM治疗组升高更明显（P<0.05）。虽然OM治疗组血清IL-4水平也升高，但差异无统计学意义（P>0.05）。（4）OM治疗组、MSC治疗组和MSC联合OM治疗组血清IL-10水平显著升高（P<0.05），且在这三组中MSC联合OM治疗组最高（P<0.05）。（5）双光子荧光成像显示，无论是否注射OM，肝脏中均未检测到MSC信号。