益气固表丸含药血清对香烟烟雾提取物诱导的3T3-L1前脂肪细胞脂质积累及炎症的影响研究  

Effect of serum containing Yiqi Gubiao pills on lipid accumulation and inflammation of 3T3-L1 preadipocytes induced by CSE

作　　者：梁倩倩 陈乙菲 李翔鹏 李风森[3] 李争 LIANG Qianqian;CHEN Yifei;LI Xiangpeng;LI Fengsen;LI Zheng(The Fourth Clinical Medical College,Xinjiang Medical University,Urumqi 830099,China)

机构地区：[1]新疆医科大学第四临床医学院,新疆乌鲁木齐830099 [2]新疆医科大学中医学院,新疆乌鲁木齐830017 [3]新疆医科大学附属中医医院,新疆乌鲁木齐830099

出　　处：《陕西中医》2024年第10期1305-1310,共6页Shaanxi Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(82060803);新疆维吾尔自治区自然科学基金资助项目(2022D01C724);新疆维吾尔自治区科学技术厅青年拔尖人才项目[青年科技创新人才(2023TSYCCX0063)]。

摘　　要：目的:通过体外实验观察益气固表丸含药血清(YQGB)对香烟烟雾提取物(CSE)诱导的3T3-L1前脂肪细胞脂质积累及炎症的影响。方法:诱导3T3-L1前脂肪细胞分化,油红O染色及试剂盒检测分析细胞内脂质积累水平,使用CCK-8检测分化过程中不同浓度CSE、终浓度为CSE 2%及不同浓度的益气固表丸含药血清、过氧化物酶体增殖物激活受体γ(PPARγ)激动剂(罗格列酮,ROS)对细胞增殖能力的影响;采用Western blot和RT-qPCR法测定PPARγ、CCAAT增强子结合蛋白α(C/EBPα)、甾醇调节元件结合蛋白-1c(SREBP-1c)、白细胞介素-6(IL-6)、肿瘤坏死因子α(TNF-α)的蛋白及mRNA的表达量。采用ELISA法检测细胞培养上清IL-6、TNF-α的表达量。结果:油红O染色显示,3T3-L1前体脂肪细胞10 d后已诱导为完全成熟的脂肪细胞;与对照组相比,CSE组油红O染色出现明显的脂滴积累,并且总胆固醇(TC)和甘油三酯(TG)含量明显增加,益气固表丸含药血清可以降低TC、TG的含量,且其抑制作用呈浓度依赖性(P<0.05)。Western blot和RT-qPCR结果显示,益气固表丸含药血清以浓度依赖的方式降低PPARγ、C/EBPα、SREBP-1c、IL-6、TNF-α蛋白及mRNA表达(P<0.05)。ELISA检测显示,益气固表丸含药血清可降低IL-6、TNF-α表达,且其抑制作用呈浓度依赖性(P<0.05)。结论:益气固表丸含药血清可以缓解CSE诱导3T3-L1脂肪细胞的脂质积累,抑制细胞炎症因子分泌,发挥抑炎作用。Objective:To observe the effect of Yiqi Gubiao pills drug serum(YQGB)on 3T3-L1 preadipocyte lipid accumulation and inflammation induced by cigarette smoke extract(CSE)in vitro.Methods:Differentiation of 3T3-L1 preadipocytes was induced,lipid accumulation in the cells was detected by oil red O staining and assay kit.The effects of different concentrations of CSE during differentiation,the final concentration of CSE 2%,the serum containing Yiqi Gubiao pills and the PPARγagonist(Rosiglitazone,ROS)on cell proliferation were detected by CCK-8.Western blot and RT-qPCR were used to detect the activated receptor of oxisome proliferator of protein and mRNA expression levels ofγ(PPARγ),CCAAT enhancer binding proteinα(C/EBPα),sterol regulatory element-binding protein-1c(SREBP-1c),interleukin-6(IL-6),tumor necrosis factorα(TNF-α).The expression of PPARγwas observed by immunofluorescence,and the expression of IL-6 and TNF-αwere detected by ELISA.Results:Oil red O staining showed that 3T3-L1 precursor adipocytes were induced into fully mature adipocytes after 10 days.Compared with the control group,oil red O staining showed obvious accumulation of lipid droplets,and the contents of total cholesterol(TC)and triglyceride(TG)were significantly increased in the CSE group.The contents of TC and TG were decreased in the serum containing Yiqi Gubiao pills,and the inhibitory effect was concentration-dependent(P<0.05).Western blot and RT-qPCR results showed that the expression of PPARγ,C/EBPα,SREBP-1c,IL-6 and TNF-αprotein and mRNA in serum containing Yiqi Gubiao pills decreased in a concentration-dependent manner(P<0.05).ELISA results showed that the expression of IL-6 and TNF-αin serum of Yiqi Gubiao pill was decreased in a dose-dependent manner,and the inhibitory effect was dose-dependent(P<0.05).Conclusion:The serum containing Yiqi Gubiao pills can relieve the accumulation of lipid in 3T3-L1 adipocytes induced by CSE,inhibit the secretion of inflammatory cytokines,and play an anti-inflammatory role.

关 键 词：慢性阻塞性肺疾病 益气固表丸含药血清 脂肪积累 炎症因子 过氧化物酶体增殖物激活受体Γ 总胆固醇 

分 类 号：R563[医药卫生—呼吸系统]