南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (5): 705-711.doi: 10.12122/j.issn.1673-4254.2022.05.11

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TSPAN8参与小鼠非酒精性脂肪性肝病的脂质代谢

张 佳，薛 薇，章述军，朱雅莉，阳 成，高 月，石凌枫，黄文祥   

  1. 重庆医科大学附属第一医院感染科，重庆 400016
  • 出版日期:2022-05-20 发布日期:2022-06-02

TSPAN8 is involved in lipid metabolism in non-alcoholic fatty liver disease in mice

ZHANG Jia, XUE Wei, ZHANG Shujun, ZHU Yali, YANG Cheng, GAO Yue, SHI Lingfeng, HUANG Wenxiang   

  1. Department of Infectious Diseases, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
  • Online:2022-05-20 Published:2022-06-02

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摘要： 目的 探讨四跨膜蛋白8（TSPAN8）在非酒精性脂肪性肝病（NAFLD）发生发展中的变化及其在脂质代谢中的作用。方法 将30只C57BL/6J 雄性小鼠随机分为普通饲料组（ND，n=15）和高脂饲料组（HFD，n=15）。Western blot检测1、3、6月末各组小鼠肝脏组织中TSPAN8的表达水平。HepG2细胞分成6组：完全培养基培养，无其他处理的命名为对照组（CON）；游离脂肪酸（FFA）处理以构建NAFLD细胞模型的命名为FFA组；TSPAN8过表达细胞命名为PCDNA-TSPAN8组，其对照为PCDNA3.1 组；FFA处理后的PCDNA-TSPAN8命名为PCDNA-TSPAN8+FFA组，其对照为PCDNA3.1+FFA组；qRT-PCR及Western blot检测CON组和FFA组细胞中TSPAN8表达水平；油红O染色法和全自动生化仪检测PCDNA-TSPAN8+FFA和PCDNA3.1+FFA组细胞内脂质蓄积情况。qRT-PCR检测与脂质代谢相关基因mRNA的表达情况。结果 Western blot显示，与同喂养时间ND组相比，HFD喂养3、6月的小鼠肝组织中TSPAN8的表达下降（P<0.05）。qRT-PCR与Western blot显示，与CON组比较，TSPAN8在FFA组中的表达下降（P<0.01）。与PCDNA3.1+FFA组相比，PCDNA-TSPAN8+FFA组中细胞内甘油三酯（TG）水平下降（P<0.001）。qRT-PCR示，与PCDNA3.1组比较，脂肪酸转运蛋白 5（FATP5）在PCDNA-TSPAN8组中表达降低（P<0.01），与CON组相比，FATP5在FFA组表达上调（P<0.001）。结论 TSPAN8参与NAFLD的脂质代谢，细胞过表达TSPAN8可能通过降低FATP5的表达来抑制细胞内脂质的沉积，可能具有潜在的临床价值。

关键词: 非酒精性脂肪性肝病；四跨膜蛋白8；脂代谢

Abstract: Objective To investigate the changes of tetraspanin 8 (TSPAN8) expression levels and its role in lipid metabolism during the development of non-alcoholic fatty liver disease (NAFLD). Methods Thirty male C57BL/6J mice were randomly divided into normal diet group and high-fat diet (HFD) group (n=15), and after feeding for 1, 3, and 6 months, the expression levels of TSPAN8 in the liver tissues of the mice were detected with Western blotting. In a HepG2 cell model of NAFLD induced by free fatty acids (FFA), the effect of TSPAN8 overexpression on lipid accumulation was examined using Oil Red O staining and an automated biochemical analyzer, and the mRNA expressions of the key genes involved in lipid metabolism were detected using qRT-PCR. Results Western blotting showed that compared with that in mice with normal feeding, the expression of TSPAN8 was significantly decreased in the liver tissues of mice with HFD feeding for 3 and 6 months (P<0.05). In HepG2 cells, treatment with FFA significantly decreased the expression of TSPAN8 at both the mRNA and protein levels (P< 0.01). TSPAN8 overexpression in FFA-treated cells showed significantly lowered intracellular triglyceride levels (P<0.001) and obviously reduced mRNA expression of fatty acid transport protein 5 (FATP5) (P<0.01). The expression of FATP5 was significantly increased in FFA- treated cells as compared with the control cells (P<0.001). Conclusion TSPAN8 is involved in lipid metabolism in NAFLD, and overexpression of TSPAN8 may inhibit cellular lipid deposition by reducing the expression of FATP5.

Key words: non-alcoholic fatty liver disease; tetraspanin 8; lipid metabolism