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目的研究肝再生增强因子(ALR)保护急性肝损伤的作用及机制。方法将30只BALB/c小鼠随机分为正常对照组、急性肝损伤组和ALR干预组。急性肝损伤组小鼠按2 m L/kg体质量的剂量予以腹腔注射500 m L/L四氯化碳(CCl4)矿物油溶液1次。ALR干预组于CCl4注射前8 h尾静脉注射ALR质粒,正常对照组注射等量生理盐水注射液。收集小鼠肝组织和血液标本,HE染色观察肝组织病理形态学变化;生化法检测血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)水平;流式细胞术检测肝组织中调节性T细胞(Treg)的数量,实时定量PCR(qRT-PCR)检测肝组织Foxp3、ALR、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)的mRNA水平。结果 ALR干预组小鼠肝组织中ALR mRNA表达水平显著高于急性肝损伤组和正常对照组,急性肝损伤组与正常对照组相比无明显差异;流式细胞术检测结果显示ALR干预组小鼠肝组织中CD25+Foxp3+Treg/CD4+T细胞为(5.90±0.10)%,高于急性肝损伤组的(4.23±0.46)%和正常对照组的(2.93±0.74)%,急性肝损伤组显著高于正常对照组;ALR干预组小鼠肝组织中Foxp3 mRNA表达水平高于急性肝损伤组和正常对照组,急性肝损伤组高于正常对照组,但差异无统计学意义;ALR干预组与急性肝损伤组相比,IL-6 mRNA、TNF-αmRNA表达水平降低,而与正常对照组相比无明显变化,急性肝损伤组与正常对照组相比,IL-6 mRNA、TNF-αmRNA表达水平明显升高。结论 ALR通过上调Treg数量保护小鼠急性肝损伤,可能与Treg下调肝脏IL-6、TNF-α表达有关。
Objective To study the role and mechanism of liver regeneration augmentation factor (ALR) in protecting acute liver injury. Methods Thirty BALB / c mice were randomly divided into normal control group, acute liver injury group and ALR intervention group. Acute liver injury mice were injected intraperitoneally with 500 m L / L CCl4 mineral oil solution at a dose of 2 m L / kg body weight once. The ALR plasmid was injected into the caudal vein 8 h before CCl4 injection in the ALR intervention group, and the normal control group was injected with the same amount of saline injection. The liver tissues and blood samples of mice were collected and the histopathological changes of liver tissues were observed by HE staining. The level of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) The number of regulatory T cells (Tregs) in liver tissues and the mRNA levels of Foxp3, ALR, IL-6 and TNF-α in liver tissues were detected by qRT-PCR. . Results The expression of ALR mRNA in liver of ALR intervention group was significantly higher than that in acute liver injury group and normal control group, but there was no significant difference between acute liver injury group and normal control group. The results of flow cytometry showed that ALR intervention group The percentage of CD25 + Foxp3 + Treg / CD4 + T cells in murine liver tissue was (5.90 ± 0.10)% higher than that in acute liver injury group (4.23 ± 0.46)% and in normal control group Group was significantly higher than the normal control group; liver tissue ALR intervention group Foxp3 mRNA expression levels higher than the acute liver injury group and the normal control group, acute liver injury group than the normal control group, but no significant difference; ALR intervention Compared with the normal control group, the levels of IL-6 mRNA and TNF-αmRNA in the group of acute liver injury were lower than those in the control group αmRNA expression was significantly increased. Conclusion ALR protects mice from acute liver injury by increasing the number of Tregs, which may be related to the down-regulation of hepatic IL-6 and TNF-α expression by Tregs.