1,25(OH)_(2)D_(3)改善胆碱缺乏氨基酸饮食诱导的非酒精性脂肪性肝炎大鼠模型肝脏炎症的机制分析  

Mechanism of 1,25(OH)_(2)D_(3) improving liver inflammation in a rat model of nonalcoholic steatohepatitis induced by choline-deficient L-amino acid-defined diet

作　　者：朱海洋 崔京淑 杨柳 周梦婷 童戬 韩红梅 ZHU Haiyang;CUI Jingshu;YANG Liu;ZHOU Mengting;TONG Jian;HAN Hongmei(Department of Gastroenterology,The Affiliated Hospital of Yanbian University,Yanji,Jilin 133001,China;Department of Pathology,The Affiliated Hospital of Yanbian University,Yanji,Jilin 133001,China)

机构地区：[1]延边大学附属医院消化内科,吉林延吉133001 [2]延边大学附属医院病理科,吉林延吉133001

出　　处：《临床肝胆病杂志》2025年第2期254-262,共9页Journal of Clinical Hepatology

基　　金：国家自然科学基金(81860110)。

摘　　要：目的分析1,25(OH)_(2)D_(3)对非酒精性脂肪性肝炎(NASH)大鼠模型肝脏过氧化物酶体增殖物激活受体γ(PPAR-γ)水平以及肝巨噬细胞表型、肝脏炎症的影响,初步探讨1,25(OH)_(2)D_(3)改善NASH肝脏炎症的相关机制。方法将24只SPF级Wistar大鼠适应性喂养1周后,随机分为4组。正常组(n=6):给予胆碱充足的氨基酸(CSAA)饮食;正常+1,25(OH)_(2)D_(3)组(n=6):给予CSAA饮食+1,25(OH)_(2)D_(3)干预;模型组(n=6):给予胆碱缺乏的氨基酸(CDAA)饮食;模型+1,25(OH)_(2)D_(3)组(n=6):给予CDAA饮食+1,25(OH)_(2)D_(3)干预。1,25(OH)_(2)D_(3)剂量为5μg/kg,经腹腔注射,每周2次,共12周。检测每组大鼠血清AST、ALT水平;观察肝组织病理学严重程度,评估SAF评分。检测肝组织M1型、M2型肝巨噬细胞,分析肝巨噬细胞表型变化;并检测肝组织TNF-α、IL-1β、IL-4、IL-10水平以及PPAR-γ的mRNA水平。组间比较采用双因素方差分析,进一步比较采用LSD-t多重检验,相关性分析采用Pearson相关法。结果与正常组比较,CDAA饮食诱导的NASH模型AST和ALT水平均升高(P值分别为0.019、<0.001),肝组织病理学SAF评分(P<0.001)、M1型肝巨噬细胞表达水平(P<0.001)、M1/M2表型比值(P=0.001)、TNF-α表达水平显著增加(P<0.001),IL-4表达水平降低(P=0.025);1,25(OH)_(2)D_(3)显著降低了CDAA饮食诱导NASH大鼠血清ALT水平(P<0.001)、肝组织病理学SAF评分(P<0.001)、M1型肝巨噬细胞水平(P<0.001)、M1/M2型比值(P=0.001)、IL-1β水平(P<0.001),增加了M2型肝巨噬细胞水平(P=0.017)、IL-10水平(P=0.039)、IL-4水平(P<0.001)、PPAR-γ水平(P=0.016)。CDAA饮食诱导的NASH模型和1,25(OH)_(2)D_(3)在大鼠血清AST和ALT水平(P值分别为0.007、0.008)、肝组织病理学SAF评分(P<0.001)、M1型肝巨噬细胞水平(P<0.001)、M2型肝巨噬细胞水平(P=0.008)、M1/M2表型比值(P=0.005)、TNF-α水平(P<0.001)、IL-10水平(P=0.038)、IL-4水平(P<0.001)、PPAR-γ水平(P=0.009)中有显著交互作用。相关性�Objective To investigate the effect of 1,25(OH)_(2)D_(3) on the level of peroxisome proliferator-activated receptor-γ(PPAR-γ)in the liver,the phenotype of hepatic macrophages,and liver inflammation in a rat model of nonalcoholic steatohepatitis(NASH),as well as the mechanism of 1,25(OH)_(2)D_(3) improving liver inflammation.Methods After 1 week of adaptive feeding,24 specific pathogen-free Wistar rats were randomly divided into normal group[choline-supplemented L-amino acid-defined(CSAA)diet],normal+1,25(OH)_(2)D_(3) group[CSAA diet+1,25(OH)_(2)D_(3)],model group[choline-deficient L-amino acid-defined diet(CDAA)diet],and model+1,25(OH)_(2)D_(3) group[CDAA diet+1,25(OH)_(2)D_(3)],with 6 rats in each group.The dose of 1,25(OH)_(2)D_(3) was 5μg/kg for intraperitoneal injection twice a week for 12 weeks.The serum levels of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)were measured,liver histopathology was observed,and SAF score was assessed.M1 hepatic macrophages and M2 hepatic macrophages were measured to analyze in the change in the phenotype of hepatic macrophages,and ELISA was used to measure the levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-4(IL-4),and interleukin-10(IL-10)in liver tissue,and qPCR was used to measure the mRNA level of PPAR-γ.The two-factor analysis of variance was use for comparison between groups,and the least significant difference t-test was used for further comparison;the Pearson method was used for correlation analysis.Results Compared with the normal group,the model rats with CDAA diet-induced NASH had significant increases in the serum levels of AST and ALT(P=0.019 and P<0.001),the SAF score of liver histopathology(P<0.001),the level of M1 hepatic macrophages(P<0.001),and the ratio of M1 and M2 hepatic macrophages(P<0.001),as well as a significant increase in the level of TNF-α(P<0.001)and a significant reduction in the level of IL-4 in liver tissue(P=0.025).The 1,25(OH)_(2)D_(3) group had significant reductions in the serum levels

关 键 词：非酒精性脂肪性肝炎 骨化三醇 炎症 PPARΓ 大鼠 Wistar 

分 类 号：R57[医药卫生—消化系统]