摘要
目的:研究丹参对非酒精性脂肪肝(NAFLD)大鼠Th17细胞及相关细胞因子的影响,为临床上丹参治疗NAFLD提供实验依据。方法:SPF级别SD大鼠32只,随机分为模型组、空白对照组、葛根对照组、丹参组,每组8只。模型组给予高脂饲料喂养4周以建立NAFLD模型;丹参治疗组及葛根对照组大鼠给予6.25g·kg-1·d-1的浓缩液每日1次灌胃;除了空白对照组外,其余各组大鼠均给予高脂饲料喂饲4周。实验完全结束(第4周),各组大鼠禁食12~14 h,禁水2 h后,采血,收集肝脏组织,检测各组大鼠血脂(TC、TG、LDL-C、HDL-C)水平及肝功能(AST、ALT)、肝脏指数,观察肝组织病理学改变。流式细胞仪检测外周血Th17、Treg细胞含量。通过ELISA法检测血清IL-6、IL-17、TNF-α水平;RT-PCR法检测肝脏组织RORγt基因表达。结果:连续4周喂养高脂饮食后,模型组大鼠肝脏发生脂肪变性,有大量炎症细胞浸润。与模型组相比,丹参治疗组、葛根对照组大鼠TC、TG、LDL-C、ALT、AST水平及肝脏指数明显低于模型组(P<0.05),HDL-C水平明显高于模型组(P<0.05)成功复制NAFLD大鼠模型。丹参治疗组大鼠外周血Th17细胞含量、IL-6、IL-17水平、大鼠RORγt基因表达量显著低于模型组及葛根对照组(P<0.05);TNF-α水平低于模型组及葛根对照组;Treg细胞含量高于模型组(P<0.05)及葛根对照组(P>0.05);Treg/Th17显著高于模型组及葛根对照组(P<0.05)。结论:丹参通过降低血清中IL-6、IL-17、TNF-α水平,抑制RORγt基因表达,降低外周血Th17细胞含量,升高Treg细胞含量,调整Th17/Treg平衡,从而抑制NAFLD发生发展。
Objective:To study the effects of Salvia mil on Th17 cells and related cytokines in non-alcoholic fatty liver disease(NAFLD)rats.Methods:SD rats were randomly divided into control group,model group,Pueraria control group and Salvia mil treated group(8 rats in each group).Rats in Salvia mil treated group and Pueraria control group were treated with Salvia mil or Pueraria at the dose of 6.25g·kg-1·d-1 by gavage once a day for 4 weeks.Except for control group,rats in other groups were fed with high-fat diet for 4 weeks.The levels of serum lipids(TC,TG,LDL-C,HDL-C),liver function indexes(AST,ALT)and liver index were measured,and the pathological changes of liver tissues were observed.Th17 and Treg cells in peripheral blood were detected by flow cytometry.Serum levels of IL-6,IL-17 and TNF-alpha were detected by ELISA.The expression level of RORγt gene in liver tissue was detected by RT-PCR.Results:NAFLD rat model was successfully established by feeding high-fat diet for 4 weeks.The liver of model group had steatosis and a large number of inflammatory cells infiltrated.Compared with model group,the levels of TC,TG,LDL-C,ALT,AST and liver index in Salvia mil treated group and Pueraria control group were significantly lower than those of model group(P<0.05),and the level of HDL-C was significantly higher than that of model group(P<0.05).The content of Th17 cells in peripheral blood of rats in Salvia mil treated group was 1.58%±0.18%,significantly lower than that of model group 3.50%±0.16%and Pueraria control group 1.69%±0.20%(P<0.05),Treg cell content was 4.58%±0.18%,higher than that of model group 3.71%±0.22%(P<0.05)and Pueraria control group 4.52%±0.19%(P>0.05),Treg/Th17 was 2.91±0.25,significantly higher than that of model group 1.06±0.24 and Pueraria control group 2.67±0.26(P<0.05).The levels of IL-6,IL-17 and TNF-αin Salvia mil treated group were lower than those of model group and Pueraria control group,and the levels of IL-6 and IL-17 were decreased significantly(P<0.05).The expression level of RORγt gene in Salvia mil treated group was 0.72±0.09,significantly lower than that of model group 2.16±0.14 and Pueraria control group 1.32±0.08(P<0.05).Conclusion:Salvia mil can decrease the levels of IL-6,IL-17 and TNF-α,inhibit the expression of RORγt gene,decrease the content of Th17 cells,increase the content of Treg cells,adjust the balance of Th17/Treg,then inhibit the development of NAFLD.
作者
崔桂玉
白剑
苗兰英
林大勇
王学良
刘喜成
CUI Gui-yu;BAI Jian;MIAO Lan-ying;LIN Da-yong;WANG Xue-liang;LIU Xi-cheng(Medical School of Inner Mongolia University for Nationalities,Tongliao 028000;Liao Ning University of Traditional Chinese Medicine,Shenyang 110847;Department of Anesthesiology,Shenzhen People’s Hospital,Shenzhen 518020,China)
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2019年第6期517-522,共6页
Chinese Journal of Applied Physiology
基金
辽宁省自然科学基金(201101142)
辽宁省社会发展攻关计划(2012225018)。
