Th1/Th2类细胞因子的变化对急性免疫性肝损伤的影响研究

目的探讨Th1/Th2类细胞因子的变化对ConA诱导的急性免疫性肝损伤的机制,以及脾脏对急性免疫性肝损伤的影响作用。方法将Balb/c小鼠随机分为两组：正常对照组,肝损伤组。正常对照组尾静脉注射等量生理盐水,肝损伤组尾静脉注射12.5mg/Kg ConA一次。各组分别于ConA注射后8h,24h,72h取材,进行下列研究：①HE染色观察各组小鼠肝脏病理学改变。②经眼球取血,收集血清测ALT和AST。③收集各组小鼠血清及新鲜肝、脾组织（各100mg）,获取肝、脾组织裂解液。用多参数细胞因子检测技术即FlowCytomix技术,通过流氏细胞仪对荧光素PE信号强度的检测,实现对各组小鼠血清、肝组织、脾组织内多种Th1/Th2类因子的定性定量分析。结果①HE染色：正常对照组肝组织结构正常。肝损伤组8h时表现为急性肝损伤表现,24h时可见大片坏死灶,72h时肝损伤缓解。②血清ALT和AST检测：正常对照组3个时间点内无明显升高,肝损伤组3个时间段内ALT和AST均高于正常对照组,有显著性差异。③Th1/Th2细胞因子检测结果：肝脏：肝损伤组8h时Th1和Th2类细胞因子均明显升高,与正常对照组比较有显著性差异,24h后开始下降,降至正常水平或正常水平以下,呈明显下降趋势。血清：肝损伤组Th1,Th2类细胞因子8h均升高,24h后逐步降低。脾脏：肝损伤组Th1,Th2类细胞因子8h时均升高,与正常对照组比较,有显著性差异,24h时明显降低。结论①ConA诱导的急性免疫性肝损伤主要是由Th1类细胞、巨噬细胞和Th2类细胞分泌的炎性因子所造成。②脾脏通过Th1/Th2类细胞因子的分泌对急性免疫性肝损伤起到免疫调控作用。

The effect of Th1/Th2 cytokines on the mice with autoimmune acute liver injury

Objective To study the effect of Th1/Th2 cytokines on the mice with autoimmune acute liver injury.Methods The adult and healthy Balb/c mice were divided into two groups: normal control group and liver injury group.Normal control group was received single injection of same dose of saline,while liver injury group respectively by caudal vein injection of 12.5mg/Kg ConA.The serum,liver and spleen tissues were collected at 8h,24h and 72h after ConA injection and the following steps were tested: ① The pathological analysis of liver tissues in every groups were studied by Hematoxylin-eosin staining,② The level of ALT and AST in serum were detected.③ All the serum and the supernatant of the lysis of fresh liver(100 mg) and spleen(100 mg) tissues were used to detected the concentration and variation of multiple Th1/Th2 cytokines by Flow Cytomix technique.Flowcytometre was used to test the intensity of fluorencence.Results ①HE staining: The structure of mice liver in normal control group were normal.Liver injury can be found in the mice of liver injury group at 8h after ConA injection.At 24h,there were more edema liver cells and enlarged necrotic foci,but the congestion of sinusoids were reduced at 72h.②ALT and AST results: there are no obvious changes in ALT and AST level in the mice serum of normal control group at three time points.The concentration of ALT and AST in Liver injury group were higher than that of normal control Group.③The detection of Th1/Th2 cytokines: Liver: There were significant differences in the concentration of Th1 and Th2 between liver injury Group and normal control group at 8h.At 24h,the concentration of Th1 and Th2 was decreased to normal level or below the normal level,and appeared a downward trend.Serum: The concentration of Th1 and Th2 type cytokines in liver injury group were higher at 8h,compared with the normal control group and decreased gradually after 24h.Spleen: All the Th1 and Th2 type cytokines in liver injury group was increased at 8h compared with the normal control group,but significantly decreased after 24h.Conclusion 1.The cytokines secreted by Th1 lymphocytes,macrophage and Th2 lymphocytes were involved in acute immunological liver injury.2.The spleen can play an immunoregulated role to acute immunological liver injury by the secretion of Th1/Th2 cytokines.