岩黄连总碱提取物对小鼠免疫性肝损伤的改善作用

Improving effect of total alkaloids extract from Corydalis thalictrifolia Franch. on immune hepatic injury in mice

目的探讨岩黄连总碱提取物(CTFTAE)对小鼠免疫性肝损伤的改善作用及其可能机制。方法小鼠尾静脉iv给予卡介苗和脂多糖制备免疫性肝损伤小鼠模型。注射卡介苗当天ip给予CTFTAE0.26,0.78和2.34mg·kg-1,每天1次,共12d。末次给药后尾静脉iv给予脂多糖(7.5μg),16h后检测小鼠血清谷丙转氨酶(GPT)和谷草转氨酶(GOT)活性;HE染色,光镜下观察肝组织病理变化;ELISA法检测血清干扰素γ(IFN-γ)和白细胞介素4(IL-4)的含量。[3H]TdR掺入法测定CTFTAE体外对小鼠脾淋巴细胞增殖反应的影响。结果模型组小鼠血清GPT和GOT活性较正常对照组明显升高;CTFTAE0.26,0.78和2.34mg·kg-1剂量组血清GPT活性较模型组明显降低,GOT活性无明显变化。模型组可见明显的肝细胞溶解坏死,而CTFTAE0.78和2.34mg·L-1组肝细胞变性、坏死及炎症细胞浸润均明显减轻。模型组小鼠血清IFN-γ和IL-4含量升高,IL-4升高较明显,IFN-γ/IL-4比值降低;CTFTAE0.26,0.78和2.34mg·kg-1组IL-4含量降低,IFN-γ/IL-4比值升高。CTFTAE15～1500mg·L-1体外单独应用可增强小鼠脾脏淋巴细胞增殖反应,亦可增强刀豆蛋白A诱导的脾淋巴细胞增殖反应。结论CTFTAE对免疫性肝损伤具有改善作用,其机制可能与其调节辅助性T细胞(Th)1/Th2平衡有关。

AIM To investigate the improving effect of Corydalis thalictrifolia Franch. total alkaloids extract（CTFTAE） on immune hepatic injury in mice and its possible mechanism. METHODS The mouse model with immune hepatic injury was prepared by injecting bacillus Calmette-Guerin （BCG） and lipopolysaccharities （LPS） through tail veins. On the same day when BCG injected, CTFTAE（0.26, 0.78 and 2.34 mg·kg^-1, respectively） was ip given, once daily, for 12 d. Then LPS （7.5 μg） were injected through tail veins on the last day of CTFTAE treatment. After 16 h, the activi- ties of glutamic-pyruvic transaminase （GPT） and glutamic-oxaloacetic transaminase （ GOT ） were determined in serum, and the pathological changes in liver tissue were observed under light microscope. The contents of interferon-γ （IFN-γ） and interleukin-4 （ IL-4 ） in serum were detected by ELISA. In addition, the effect of CTFTAE on splenocyte proliferation in vitro was observed by [ 3H ] TdR incorporation assay. RESULTS The activities of GPT and GOT in serum were obviously increased in model group than that in normal control group. Compared with model group, GPT activity obviously decreased in CTFTAE 0.26, 0.78 and 2.34 mg·kg^-1 groups, but GOT activity had no significant change. It was observed that some liver cells obviously dissolved and necrotized in model group, and CTFTAE 0.78 and 2.34 mg·kg^-1 alleviated the hepatic cell degeneration, necrosis and inflammatory cell infiltration. The serum contents of INF-γ/ and IL-4 increased in model group compared with normal group, and IL-4 content increased more significantly, and the IFN-γ/IL-4 declined. CTFTAE 0.26, 0.78 and 2.34 mg·kg^-1 reduced the content of IL-4, and elevated the ratio of IFN-γ/IL-4. Furthermore, CTFTAE 15 -1500 mg·L-1 could increase splenocyte proliferation in vitro both used alone and combined with concanavalin A. CONCLUSION CTFTAE can alleviate the immune hepatic injury, which may be related with its regulation of the balance between helper T cells （Th）1 and Th2.