NASH患者外周血乏氧诱导因子1a-反义RNA 1和自噬基因Beclin1变化及其临床意义

目的研究非酒精性脂肪性肝炎(NASH)患者外周血单个核细胞(PBMC)乏氧诱导因子1a-反义RNA 1(HIF1a-AS1)和血清自噬基因beclin1水平变化及其临床意义。方法2019年1月~2021年4月我院诊治的NASH患者118例(早期纤维化、进展期纤维化和肝硬化分别为43例、46例和29例)和同期健康体检者118例,使用全自动生化分析仪检测空腹血糖(FBG)、总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C)水平,采用实时荧光定量PCR法检测PBMC HIF1a-AS1水平,采用ELISA法检测血清beclin1水平。结果NASH患者FBG、血清TC、TG、LDL-C、PBMC HIF1a-AS1和血清beclin1水平分别为(5.9±1.6)mmol/L、(6.2±0.5)mmol/L、(2.4±0.6)mmol/L、(3.7±0.9)mmol/L、(1.9±0.2)和(5.7±1.9)ng/mL,显著高于健康者【分别为(4.8±0.7)mmol/L、(5.3±0.3)mmol/L、(1.3±0.3)mmol/L、(2.3±0.6)mmol/L、(1.0±0.1)和(4.1±1.5)ng/mL,P<0.05】,而血清HDL-C水平为(1.2±0.2)mmol/L,显著低于健康者【(1.4±0.3)mmol/L,P<0.05】;肝硬化患者FBG、血清TC、TG、LDL-C、PBMC HIF1a-AS1和血清beclin1水平分别为(6.8±2.0)mmol/L、(6.8±0.8)mmol/L、(2.8±0.7)mmol/L、(4.4±1.2)mmol/L、(2.5±0.3)和(6.4±2.1)ng/mL,显著高于早期纤维化患者【分别为(5.2±1.1)mmol/L、(5.7±0.4)mmol/L、(1.9±0.5)mmol/L、(3.1±1.0)mmol/L、(1.4±0.1)和(5.1±1.3)ng/mL,P<0.05】;49例有高血压、糖尿病或/和高脂血症合并症的NASH患者FBG、血清TC、TG、LDL-C、PBMC HIF1a-AS1和血清beclin1水平分别为(6.5±1.9)mmol/L、(6.9±0.8)mmol/L、(2.7±0.8)mmol/L、(4.0±1.1)mmol/L、(2.2±0.3)和(6.3±2.0)ng/mL,显著高于69例无合并症者【分别为(5.5±1.3)mmol/L、(5.7±0.4)mmol/L、(2.2±0.5)mmol/L、(3.5±0.7)mmol/L、(1.7±0.2)和(5.3±1.6)ng/mL,P<0.05】,而血清HDL-C水平为(1.1±0.2)mmol/L,显著低于无合并症者【(1.3±0.4)mmol/L,P<0.05】。结论NASH患者PBMC HIF1a-AS1和血清beclin1水平异常升高,监测其水平可能有助于病情评估。

Inhibition of Fas/FasL mRNA expression and TNF-a release in concanavalin A-induced liver injury in mice by bicyclol

AIM: Bicyclol, 4,4'-dimethoxy-5,6,5',6'-dimethylene-dioxy-2-hydroxymethy1-2'-carbonyl biphenyl, is a new anti-hepatitis drug. The aim of the present study was to investigate the protective effect of bicyclol on concanavalin A (Con A)-induced immunological liver injury in mice and its mechanism. METHODS: Liver injury was induced by injection of Con A via tail vein of mice and assessed biochemically and histologically. Serum transaminase and tumor necrosis factor alpha (TNF-a were determined. Liver lesions were observed by light microscope. Expressions of TNF-a, interferon gamma (IFN-y), Fas and Fas ligand (FasL) mRNA in the livers were measured by RT-PCR. RESULTS: Serum transaminase level and liver lesions in Con A-induced mice were markedly reduced by oral administration of 100, 200 mg/kg of bicyclol. TNF-a level inserum was also reduced by bicyclol. Con A injection in ducedup-regulation of TNF-a, IFN-7, Fas and FasL mRNA expression in liver tissues. Bicyclol significantly down-regulated the expression of IFN-y, Fas and FasL mRNA, but only slightly affected TNF-a mRNA expression in liver tissues. CONCLUSION: Bicyclol protects against Con A-induced liver injury mainly through inhibition of Fas/FasL mRNA expression in liver tissues and TNF-a release in mice.

B7-H4对ConA诱导小鼠肝损伤保护作用的实验研究

目的探讨B7-H4对免疫性损伤肝脏的保护作用及其机制。方法将KM小鼠分为4组(每组15只):生理盐水组(A组)、pcDNA3.1-mB7-H4-Fc组(B组)、pcDNA3.1组(C组)和刀豆球蛋白A(Con A)组(D组)。B组、C组和D组动物均通过尾静脉注射Con A 25mg/kg制备肝损伤模型。在Con A注射前1d,分别给予B组每只小鼠注射pcDNA3.1-mB7-H4-Fc 100μg和C组每只小鼠注射pcDNA3.1 100μg。并于注射Con A后12h、24h和48h,各组取5只小鼠,摘眼球取血,分离血清行白介素-4(IL-4)、γ-干扰素(IFN-γ)、谷丙转氨酶(ALT)和谷草转氨酶(AST)浓度的测定;引颈处死小鼠后取肝脏,用于组织病理学检测。结果小鼠血清IL-4、IFN-γ、ALT和AST水平在注射Con A后各时点,B、C和D三组较A组均有升高(P<0.05),但B组较C组和D组减低,B组与C组间差异有统计学意义(P<0.01)。小鼠肝组织切片经HE染色,光学显微镜下可见,B组小鼠肝脏损伤均比C组和D组有不同程度的减轻。结论初步证实了B7-H4对Con A诱导的肝脏免疫性损伤具有保护作用。B7-H4可能是通过抑制IL-4和IFN-γ的产生和/或分泌而发挥对肝脏保护的作用。