染料木素磺酸钠对慢性肝损伤的保护及T淋巴细胞亚群的调节作用

The protective effects of genistein sodium sulfonate on chronic hepatic in mice by regulating the T lymphocyte subsets

目的:观察染料木素磺酸钠(genistein sodium sulfonate,GSS)对小鼠慢性肝损伤的保护作用及其对外周血T淋巴细胞亚群的调节作用。方法:取健康雄性昆明种小鼠48只,随机分成5组,分别为正常组、模型组、0.1 mg/kg GSS组、0.3 mg/kg GSS组、阳性药物对照组(2.5 mg/kg联苯双酯)。采用腹腔注射10%CCl4,体积为0.1 mL/10 g,持续6周,制备小鼠慢性肝损伤动物模型。各组分别灌胃给予不同剂量的GSS、阳性药物或生理盐水,连续6周。测定小鼠血清天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)活性,并计算AST/ALT比值;HE染色,观察肝组织形态;流式细胞术检测CD4+及CD8+T淋巴细胞亚型,并计算CD4+/CD8+T淋巴细胞比值。结果:(1)模型组小鼠肝组织可见大量肝细胞呈气球样变、脂肪变性、胞质凝聚、或肝细胞变性坏死;GSS实验组肝组织未见明显的脂肪变性或炎性病灶。(2)模型组小鼠血清AST及ALT含量升高,GSS实验组小鼠血清AST、ALT含量明显低于模型组。(3)模型组小鼠CD3+细胞比例升高,CD8+T细胞比例降低,CD4和CD8双阴性T淋巴细胞比例升高,CD4+/CD8+T淋巴细胞比值升高;GSS治疗后,CD3+细胞比例降低,CD8+T淋巴细胞比例升高,CD4+/CD8+T淋巴细胞比值降低,CD4和CD8双阴性T淋巴细胞比例降低。结论:GSS对CCl4诱导小鼠慢性肝损伤有保护作用,其机制可能通过调节T淋巴细胞亚群,影响免疫功能实现的。

Objective ：To explore the protective effects of genistein sodium sulfonate （GSS） on chronic hepatic injury in- duced by carbon tetrachloride（ CC14 ） in mice and investigate the mechanism about regulating the T lymphocyte subsets. Methods ：48 healthy Kunming male mice were randomly divided in 5 groups, which included control group, model group, bifendate（DDB） positive control group（2.5 mg/kg） , low and high dose GSS groups（0.1 mg/kg, 0.3 mg/kg）. The chronic hepatic injury mice were made by intraperitoneal administrated 10 % CC14 plant oil solution twice a week, and sustained for 6 weeks. At the same time, the mice were treated with normal saline, DDB（2.5 mg/kg） and GSS（0.1 mg/kg, 0.3 mg/kg） respectively by ig one time a day and continued for six weeks. After the last administration, the mice blood and liver were taken. Flow cytometry was used to detect T lymphocyte subsets, enzymes analysis technique was used to observe the liver function, HemateinEosin stain was used to explore the changes in liver morphology. Results ： In chronic hepatic injury mice, liver cells show ballooning change, fatty degeneration, cytoplasm condensation, degenera- tion and necrosis, the activities of ALT and AST were increased obviously, CD3 ＋ cell populationwere elevated, CD8 ＋ T lymphocyte subsets ratio was reduced, CD4 and CD8 double negative T lymphocyte cell ratio and CD4 ＋/CD8＋ ratio wereenhanced. After treated with GSS these changes were reversed. Conehtsion：GSS plays a protective role in chronic hepatic injury mice induced by CC14, and its mechanism probably result of GSS modulating the immunologic function by regula- ting the T lymphocyte subsets.