目的 建立硫代乙酰胺(TAA)诱导的大鼠急性肝损伤模型,探讨Norrin/Frizzled-4在肝损伤间质重建中的作用.方法 随机将32只SD大鼠分为正常对照组(N组)8只和TAA诱导组(M组)24只.建立TAA诱导的肝损伤大鼠模型.采用ELISA法检测血清CD105和血管内皮细胞生长因子(VEGF)水平,采用Western blot法检测原代肝星状细胞和肝组织 Norrin/Frizzled-4信号通路中细胞外配体蛋白 Norrin 和细胞膜特异性受体蛋白Frizzled-4的表达.结果 24只TAA诱导组大鼠死亡11只(45.8%);病理学检查显示急性肝损伤大鼠模型建立成功;对照组血清CD105水平为(2.18±0.05)ng/mL,显著低于TAA处理1 w组的(2.36±0.07)ng/mL或2 w组的(2.42±0.03)ng/mL,差异均有显著性(P<0.05);对照组血清VEGF为(61.48±0.39)pg/mL,显著低于模型组的(64.52±0.25)pg/mL,差异均有显著性(P<0.01);原代肝星状细胞和正常肝组织不表达Norrin蛋白或Frizzled-4蛋白,或表达量很低,急性肝损伤大鼠肝组织Norrin/Frizzled-4表达显著高于正常对照组,差异均有显著性(P<0.01).结论 急性肝损伤大鼠肝组织Norrin/Frizzled-4蛋白表达上调,可能与急性肝损伤初期维持血管网络形态及肝间质重建有关.%Objective To investigate the roles of Norrin/Frizzled-4 in the interstitial remodeling of acute liver injury. Methods 32 healthy male rats were randomly divided into control(n=8)and model group(n=24). The model of acute liver injury was established by thioacetamide(TAA)injuction. Western blot was used to detect the expression of Norrin/Frizzled-4 protein in liver tissues,and enzyme-linked immunosorbent assay was performed to measure serum CD105 and vascular endothelial growth factor(VEGF)levels. Results There were 11(45.8%) out of 24 rats died in model group during model establishment. Hematoxylin-eosin staining showed that the acute liver injury model was successfully established. Serum CD105 level was(2.18 ±0.05)ng/mL and VEGF was (61.48±0.39)pg/mL in control group,significantly lower than [(2.36±0.07)ng/mL and [(62.09±0.54)pg/mL,P<0.05] in TAA-treated for one week,or [2.42±0.03)ng/mL and(64.52±0.25)pg/mL,P<0.05] in TAA-treated for two week;Western blot analysis showed that the expressions of Norrin/Frizzled-4 proteins in model group were higher than in normal control group or in primary hepatic stellate cells(P<0.01). Conclusion The expressions of Norrin/Frizzled-4 proteins increase in liver tissues of rats with acute liver injury,which might be related to the maintaing of vascular network morphology and interstitial remodeling.
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