摘要
目的探索维生素D (VD)缺乏对酒精性肝损伤小鼠的影响及其相关作用机制。方法 40只6w龄ICR雄鼠随机分为标准组和VD缺乏组。标准组喂养标准固体饲料,VD缺乏组喂养VD缺乏固体饲料。喂养4w后再次分组,将标准组随机分成对照组(Ctrl)和酒精组(EtOH),VD缺乏组随机分成VD缺乏组(VDD)和VD缺乏+酒精组(VDD+EtOH)。Ctrl组和EtOH组喂养分别喂养Liber-DeCarli对照液体饲料和Liber-DeCarli酒精液体饲料,VDD组和EtOH+VDD组分别喂养VD缺乏对照液体饲料和VD缺乏酒精液体饲料。喂养10d后,EtOH组和EtOH+VDD组小鼠一次性给予5g/kg酒精灌胃。于给药12h后,取血清和肝组织,称量肝脏重量,计算肝脏系数;采用放射免疫法检测血清25(OH)D水平;检测血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)和天冬氨酸氨基转移酶(aspartate aminotransferase,AST)水平;检测肝脏甘油三酯含量(triglycerides,TG);肝脏HE染色观察组织病理学改变;反转录酶-聚合酶链锁反应(reverse transcription-polymerase chain reaction,RT-PCR)检测肝脏肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6 (interleukin-6,IL-6)、转化生长因子-β(transforming growth factor-β,TGFβ)和环氧合酶-2(cyclooxygenase-2,COX2)及单核细胞趋化因子-1 (monocyte chemotactic protein 1,MCP-1)和角蛋白趋化因子(keratinocyte chemoattractant,KC)的mRNA表达;Western blot检测肝脏核因子(nuclear factor-kappa B,NF-κB)p65和NF-κB p50蛋白表达。结果与EtOH相比,VDD+EtOH组小鼠肝脏指数、ALT及AST水平均明显升高;炎性因子TNF-α、IL-6、TGFβ和COX2及细胞趋化因子MCP1和KC的mRNA水平均显著上升,同时NF-κB p65和NF-κB p50蛋白表达水平均显著增加。结论 VD缺乏加重酒精性肝损伤,其作用机制可能与激活NF-κB信号通路,促进炎症反应有关。[营养学报,2020,42(5):465-471]
Objective To evaluate the effects of vitamin D deficiency on alcohol-induced liver injury in mice and explore the underlying mechanism.Methods Forty six-weeks-old male ICR mice were randomized into control group and vitamin D deficiency(VDD) group,and fed either a standard control diet or vitamin D deficient diet for 4 weeks.Then,mice in the normal diets group(n=20) were further divided into control group(Ctrl) and alcohol group(EtOH);and fed either a control liquid diet or alcohol liquid diet for 10 days.Meanwhile,mice in the vitamin D deficiency group(n=20) were also divided into vitamin D deficiency group(VDD) and vitamin D deficiency plus alcohol group(VDD+EtOH);and fed either vitamin D depleted control liquid diet,or vitamin D depleted alcohol liquid diet for 10 days,.The mice in EtOH group and VDD+EtOH group were administered with a single gavage of alcohol(5 g/kg body weight) after 10 days of diet intervention.All mice were sacrificed at 12 hours after alcohol challenge.Serum and liver specimens were collected immediately.Serum alanine aminotransferase(ALT),aspartate aminotransferase(AST) and hepatic triglycerides content were determined.Hepatic histopathological damage was analyzed with HE staining.Hepatic tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),transforming growth factor-β(TGF-β) cyclooxygenase-2(COX2),monocyte chemotactic protein1(MCP-1) keratinocyte and chemoattractant(KC) mRNAs were measured by reverse transcription-polymerase chain reaction(RT-PCR).The proteins of nuclear factor-kappaB(NF-κB) p65 and p50 were measured by Western blot.Results Compared with EtOH group,the liver index and serum ALT and AST levels were significantly increased in VDD+EtOH group.The mRNA levels of hepatic TNF-α,IL-6,TGF-β,COX2,MCP-1 and KC were significantly increased in VDD+EtOH group compared with the EtOH group.Moreover,the protein levels of NF-κB p65 and p50 were significantly higher in VDD+EtOH group than those in EtOH group.Conclusion Vitamin D deficiency aggravates alcohol-induced liver injury,and
作者
胡纯秋
徐德祥
王国秀
李李
王素芳
胡传来
HU Chun-qiu;XU De-xiang;WANG Guo-xiu;LI Li;WANG Su-fang;HU Chuan-lai(Department of Nutrition and Hygiene.School of Public Health,Anhui Medical University,Hefei 230032;Department of Toxicology,School of Public Health,Anhui Medical University,Hefei 230032,China)
出处
《营养学报》
CAS
CSCD
北大核心
2020年第5期465-471,共7页
Acta Nutrimenta Sinica
基金
国家自然科学基金(No.81402676)
安徽省自然科学基金(No.1508085QH187)
安徽省博士后研究人员科研活动经费资助项目(No.2016B141)。
关键词
维生素D缺乏
酒精性肝病
炎症
vitamin D deficiency
alcohol liver disease
inflammation
