上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2019, Vol. 39 ›› Issue (8): 812-.doi: 10.3969/j.issn.1674-8115.2019.08.002

• 论著·基础研究 • 上一篇    下一篇

SIRT7通过抑制内质网应激蛋白 GRP78减轻脂多糖或 D-氨基半乳糖 /脂多糖诱导的肝细胞凋亡

阮昕 1，张颖婷 1，韩可琪 1，林龙帅 2，陈晨 1，岳铭 1，王楚翘 1，孙英刚 3，赵庆华 2，贺明 1   

  1. 1. 上海交通大学基础医学院病理生理学系，细胞分化与凋亡教育部重点实验室，上海 200025；2. 上海交通大学附属第一人民医院骨科，上海 201620；3. 上海交通大学医学院附属新华医院心血管内科，上海 200092
  • 出版日期:2019-08-28 发布日期:2019-09-23
  • 通讯作者: 贺明，电子信箱：heming@shsmu.edu.cn。
  • 作者简介:阮昕（1993—），女，硕士生；电子信箱： kunxinhua123@sjtu.edu.cn。
  • 基金资助:
    国家自然科学基金（81470841）；上海市浦江人才计划（16PJ1405400，16PJ0004679）；上海市自然科学基金（19ZR1428400）；上海交通大学医工(理)交叉研究基金（YG2017MS02）

SIRT7 protecting hepatocytes LPS or D-GalN/LPS-induced apoptosisattenuating endoplasmic reticulum stress via inactivation of GRP78

RUAN Xin1, ZHANG Ying-ting1, HAN Ke-qi1, LIN Long-shuai2, CHEN Chen1, YUE Ming1, WANG Chu-qiao1, SUN Ying-gang3, ZHAO Qing-hua2, HE Ming1   

  1. 1. Department of Pathophysiology, Shanghai Jiao Tong University College of Basic Medical Sciences; Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Shanghai 200025, China; 2. Department of Orthopedics, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai 201620, China; 3. Department of Cardiology, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
  • Online:2019-08-28 Published:2019-09-23
  • Supported by:
    National Natural Science Foundation of China，81470841；Shanghai Pujiang Program，16PJ1405400，16PJ0004679；Natural Science Foundation of Shanghai，19ZR1428400；Medicine and Industry Crossing Project Shanghai Jiao Tong University，YG2017MS02

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摘要： 目的 ·探究 SIRT7（sirtuin 7）对脂多糖（ lipopolysaccharide，LPS）或 D-氨基半乳糖（ D-galactosamine，D-GalN）/LPS诱导的急性肝损伤的作用及机制。方法 ·将 13周龄的 C57BL/6J小鼠随机分为生理盐水组（ n5）、LPS组（n7）和 D-GalN/LPS组（n8），分别腹腔注射生理盐水、 LPS和 D-GalN/LPS。24 h后收集生理盐水组和 LPS组的小鼠血清和肝脏， D-GalN/LPS组于注射后 8 h收集血清和肝脏。利用苏木精 -伊红（ H-E）染色比较 3组小鼠肝脏病理学改变，同时观察血清学生化指标的变化；利用 TUNEL染色和 F4/80染色明确小鼠肝脏内细胞凋亡和炎症细胞浸润程度；利用 realtime-PCR检测各组小鼠肝组织中 SIRT7、白介素 -1β（interleukin 1β，IL-1β）、IL-6、肿瘤坏死因子 α（tumor necrosis factor α，TNF-α）等的 mRNA水平；利用 Western blotting检测各组小鼠肝组织中 SIRT7、激活型胱天蛋白酶 3（cleaved-caspase3）和伴侣葡萄糖调节蛋白（the 78 kDa glucose regulated protein，GRP78）等的蛋白水平。体外实验中，在正常小鼠肝细胞 AML-12细胞中过表达 SIRT7，并给予 LPS刺激，利用 Western blotting明确 SIRT7对 LPS引起的内质网应激信号通路分子的调控。结果 ·注射 LPS或 D-GalN/LPS后，小鼠肝脏出现明显的炎症细胞浸润、充血及肝细胞凋亡，血清中谷丙转氨酶（ glutamic-pyruvic transaminase，GPT）和谷草转氨酶（ glutamic-oxalacetic transaminase，GOT）水平明显升高，肝组织中 SIRT7的 mRNA和蛋白水平均明显降低，内质网应激蛋白 GRP78表达明显上调。在 AML-12细胞系中， SIRT7过表达可明显抑制 LPS引起的 GRP78 蛋白上调。结论 · SIRT7通过抑制内质网应激中的 GRP78减轻 LPS或 D-GalN/LPS诱导的肝细胞凋亡。

关键词: 去乙酰化酶（sirtuin）, 脓毒症, SIRT7, 急性肝损伤, 内质网应激, 细胞凋亡

Abstract:

Objective · To investigate the effects of SIRT7 on acute liver injury inducedlipopolysaccharide (LPS) or D-galactosamine (D-GalN)/LPS and its mechanisms. Methods · Thirteen-week-old C57BL/6J mice were randomly divided into normal saline group (n5), LPS group (n7), and D-GalN/ LPS group (n8), which were respectively intraperitoneally injected with normal saline, LPS or D-GalN/LPS. The serum and livers of normal saline group and LPS group mice were collected 24 hours after the injection, and the samples of D-GalN/LPS group were collected 8 hours after the injection. Liver pathological changes were comparedusing H-E staining, and serological indicators of the mice three groups were also compared. Liver apoptosis and inflammatory cells infiltration were determinedTUNEL staining and F4/80 staining. Meanwhile, the mRNA levels of SIRT7 and inflammatory factors, including interleukin-1β (IL-1β), IL-6, and tumor necrosis factor α (TNF-α) in livers were detectedrealtime-PCR. Western blotting was used to detect the protein levels of SIRT7, cleaved-caspase3 and the 78 kDa glucose regulated protein (GRP78) in moliver tissues. AML-12 cell line overexpressing SIRT7 was stimulated with LPS, and Western blotting was used to study the roles of SIRT7 in the endoplasmic reticulum (ER) stress inducedLPS in vitro. Results · LPS orD-GalN/LPSinduced inflammatorycellsinfiltration,hyperemia and hepatocytesapoptosis inlivers.Meanwhile, serum glutamic-pyruvic transaminase (GPT) and glutamic-oxalacetic transaminase (GOT) in the mice treatedLPS or D-GalN/LPS were significantly increased. Moreover, both liver SIRT7 mRNA and protein levels were down-regulated, while GRP78 protein in ER stress pathway was up-regulated. In AML-12 cells, SIRT7 over inhibited LPS-induced up-regulation of GRP78. Conclusion · SIRT7 protects against LPS or D-GalN/LPS-induced hepatocytes apoptosisattenuating ER stress via inactivating GRP78.

Key words: sirtuin, sepsis, SIRT7, acute liver injury, endoplasmic reticulum stress, apoptosis

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