粒细胞集落刺激因子对四氯化碳所致小鼠慢性肝损伤的治疗作用

Therapeutic effect of granulocyte colony stimulating factor on CCi4 induced chronic liver injury in mice

目的观察重组人粒细胞集落刺激因子（rhGCsF）对四氯化碳（CCl4）所致小鼠慢性肝损伤的治疗作用。方法清洁级雄性BALB／C小鼠分为治疗组与对照组。每周2次CCl4腹腔注射制备慢性肝损伤模型。造模成功后治疗组给予rhGCsF（200big·kg^-1·d^-1）皮下注射7d，对照组给予同等剂量0．9％氯化钠溶液。测定小鼠体质量、肝重和脾重。采用临床常规方法检测肝功能、肝纤维化指标。对肝纤维化程度进行评分。流式细胞仪计数分析肝组织中CD34＋细胞，免疫组化法测定Thy-1^＋表达。结果治疗组小鼠第8和15天时脾重与肝重比值（15．94％±1.20％和10．52％±0．66％）与对照组（7．14％±1.68％和8．31％±1．71％）比较差异有统计学意义（P值均〈O．05），两组小鼠体质量和肝重差异无统计学意义（P〉0．05）。第15天时治疗组小鼠白蛋白水平快速上升。第30天时治疗组丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、透明质酸（HA）、层粘连蛋白（LN）水平均低于对照组。第30天时两组肝纤维化程度计分差异有统计学意义（治疗组5．49±2．16，对照组8．74±1．86，P〈0．05）。治疗组小鼠肝脏组织中CD34＋细胞和Thy-1^＋阳性细胞数在第8天（9．54±2．24和5．10±1．25）和第15天（8．18±1．93和7．53±1．39）时高于对照组（第8天时5．40±0．99和3．25±0．75；第15天时4．46±0．77和3．35±0．86，P值均〈O．05）。结论rhG-CSF能促进慢性肝损伤的恢复，将为肝纤维化提供一个新的治疗方法。

Objective To observe the therapeutic effects of recombinant human granulocyte colony stimulating factor （rhG-CSF） on CC14 induced chronic liver injury. Methods Male BALB/C mice were randomly allocated into treatment and control groups. The mice model were established by injection with 40 % CC14 twice a week. The mice in treatment group were subcutaneously injected with rhG-CSF（200μg/kg） daily for 7 days, while the control mice were received the same volumes of saline. The mice were sacrificed to get weight, liver mass and spleen mass. The count of CD34＋ cells and Thy-1 ＋ cells were analyzed by flow cytometry and immunohistochemical staining, respectively. Results The ratio of liver/spleen was 15.94±1. 20 and 10. 52±0. 66 on day 8 and 15 in treatment group, respectively, while those were 7.14±1.68 and 8. 31 ± 1. 71 in control group, respectively（all P value〈0. 05）. But there was no significant difference in body weight and liver mass between two groups（P〉0.05） The concentration of album in treatment group was raised rapidly on day 15. The concentrations of alanlne aminotransferase （ALT）, aspartate aminotransferase （AST）, hyaluronic acid （HA） and laminin （LN） on day 30 were significantly lower in treatment group compared to control group （P 〈 0. 05）. There was significant difference in score of liver fibrosis on day 30 between two groups （trealment group： 5.49 ±2.16 ,control： 8.74!1.86, P〈0.05）. The number of CD34＋ cell and Thy-1 ; in treatment group （on day 8： 9.54±2.24 and 5.10±1.25 and on day 15： 8. 18±1.93 and 7.53±1.39, respectively） were higher than those in control group （on day 8：5.40±0.99 and 3.25±0.75;on 15 days： 4.46±0.77 and 3.35±0.86,all P value〈0.05）. Conclusion The rhC-CSF may improve the reparation of chronic liver injury, and may provide a novel metlaod in treatment of liver fibrosis.