维生素D调控Kupffer细胞极化在非酒精性脂肪性肝病防治中的作用  

Effect of vitamin D on non-alcoholic fatty liver disease in mice and its mechanism

作　　者：罗雯静 董显文 赵巧素 郭雯莹 柳惠未 叶桦[1] LUO Wenjing;DONG Xianwen;ZHAO Qiaosu;GUO Wenying;LIU Huiwei;YE Hua(Department of Gastroenterology,the Affiliated Lihuili Hospital,Ningbo University,Ningbo 315000,China)

机构地区：[1]宁波大学附属李惠利医院消化内科,315000

出　　处：《浙江医学》2023年第9期916-921,I0004,共7页Zhejiang Medical Journal

基　　金：宁波市自然科学基金资助项目(202003N4234);宁波市临床医学研究中心项目(2019A21003)。

摘　　要：目的探讨维生素D在非酒精性脂肪性肝病(NAFLD)防治中的作用及可能机制。方法30只C57BL/6小鼠按照随机数字表法分为对照组(正常饮食)、高脂饮食组(高脂饮食)和维生素D补充组[高脂饮食同时补充活性维生素D-1,25(OH)2D3],每组10只。喂饲相应食物16周后每组取5只小鼠处死后取肝组织,另5只小鼠行胶原酶原位灌注获取Kupffer细胞。采用HE染色观察肝组织病理学表现;采用real-time PCR法、Western blot法分别检测肝组织脂质合成和分解代谢相关基因、蛋白表达水平;采用real-time PCR法检测Kupffer细胞M1/M2极化基因表达水平。结果与高脂饮食组相比,维生素D补充组小鼠肝组织脂肪变性减轻,肝组织脂质合成基因固醇调节元件结合蛋白1C(SREBP1C)、脂肪酸合酶(FASN)和脂质分解基因脂酰辅酶A氧化酶l(ACOX1)、肉碱棕榈酰转移酶1A(CPT1A)mRNA表达水平均明显降低,肝组织脂质合成蛋白SREBP1C、FASN和脂质分解蛋白ACOX1、CPT1A表达水平均明显降低,Kupffer细胞M1极化基因诱生型一氧化氮合酶2(iNOS2)、TNF-α和IL-6及M2极化基因IL-10 mRNA表达水平均明显降低,差异均有统计学意义(均P<0.05)。结论补充维生素D可改善NAFLD小鼠肝脏脂肪变性,其机制可能与抑制Kupffer细胞M1型极化进而影响肝脏脂质代谢水平有关。Objective To investigate the effect and mechanism of vitamin D on non-alcoholic fatty liver disease(NAFLD).Methods Thirty wild-type C57BL/6 mice were randomly assigned into three groups with 10 animals in each group.Mice in control group were fed with normal diet,mice in HF diet group were fed with high-fat diet and mice in HF+VD group were given HF diet and 1,25(OH)2D3.After 16 weeks of feeding mice were sacrificed,liver tissues were harvested from 5 mice in each group for pathological examination with HE staining,and in situ perfusion of collagenase was performed in another 5 mice of each group to isolate Kupffer cells.The mRNA and protein expression of lipometabolic enzymes of liver tissues were detected by real-time PCR and Western blot,respectively.The M1/M2 phenotype markers of Kupffer cells were detected by real-time PCR.Results Vitamin D supplementation alleviated HF diet-induced hepatic steatosis in NAFLD.The mRNA expression of hepatic lipid-synthesis genes SREBP1C and FASN as well as lipid-decomposition genes ACOX1 and CPT1A were significantly decreased in HF+VD group compared to HF group.The protein expression of SREBP1C,FASN,ACOX1 and CPT1A were significantly decreased in HF+VD group compared to HF group.Expression of M1 gene markers,including iNOS2,TNF-α and IL-6,and M2 gene marker IL-10 of Kupffer cells in HF+VD group were significantly decreased compared to those from HF group(all P<0.05).Conclusion Vitamin D supplementation can alleviate HF diet-induced hepatic steatosis in NAFLD mice,which is associated with the reversing pro-inflammatory M1 polarization of Kupffer cells and altering lipid metabolism in liver.

关 键 词：维生素D 非酒精性脂肪性肝病 KUPFFER细胞 极化 

分 类 号：R575.5[医药卫生—消化系统]