Effect of CTRP13 regulates high glucose-induced autophagy dysfunction of primary rat liver sinusoidal endothelial cells through the AMPK/mTOR pathway
10.3969/j.issn.1006-6187.2023.12.010
- VernacularTitle:C1q/肿瘤坏死因子相关蛋白13通过单磷酸腺苷激活的蛋白激酶/哺乳动物雷帕霉素靶点复合物通路对高糖诱导的大鼠原代肝窦内皮细胞自噬功能影响的研究
- Author:
Jing YU
1
;
Qi ZHANG
;
Jing LIU
;
Zibing QIAN
;
Limin TIAN
;
Peiyun ZENG
;
Ruixia YANG
;
Jie YANG
;
Rui CUI
;
Zhengping CHANG
Author Information
1. 730000 兰州,甘肃省人民医院老年医学科
- Keywords:
Rat liver sinusoidal endothelial cells;
C1q/tumor necrosis factor-related protein 13;
AMP-activated protein kinase;
Mammalian target of rapamycin complex;
Autophagy
- From:
Chinese Journal of Diabetes
2023;31(12):929-937
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of C1q/tumor necrosis factor-related protein 13 protein(CTRP13)on the autophagy function of primary rat liver sinusoidal endothelial cells(rLSECs)induced by high glucose through AMP-activated protein kinase/mammalian target of rapamycin complex(AMPK/mTOR)pathway.Methods After isolation,identification and culture,original rat liver sinusoid endothelial cells(rrLSECs)were divided into normal control(NC)group,high glucose(HG)group,HG +LV-CTRP13 group,HG+ lentiviral empty vector(LV-Con)group(HG+LV-Con).CTRP13 lentivirus over expression vector(LV-CTRP13)and lentivirus empty vector(LV-Con)were constructed and transfected into rrLSECs.According to the intervention methods of AMPK inhibitor Compound C,mTOR inhibitor Torin1 and autophagy inhibitor 3MA,the transfected cell were divided into normal control(NC)group,high glucose(HG)group,HG+LV-CTRP13 group,HG+lentiviral empty vector(LV-Con)group(HG+ LV-Con).qRT-PCR and western blot were used to detect the mRNA and protein expression levels of CTRP13,autophagy related protein Beclin1,human microtubule-associated protein light chain 3II(LC3II),human plasma membrane membrane vesicle association proteins(PLVAP)and p-AMPK and p-MTOR in rat rLSECs of each group.Results Compared with NC group,the number of autophagosome was decreased in HG and HG+LV-CTRP13 group(P＜0.05).Compared with HG group,the number of autophagosome bodies was increased in HG +LV-CTRP13 group(P＜0.05).The CTRP13 mRNA and protein expression was higher in NC and HG + LV-CTRP13 groups than in HG and HG + LV-Con groups(P＜0.05).In HG+LC-CTRP13 group,Beclin1,LC3II,p-AMPK,and AMPK mRNA,Beclin1,LC3II/LC3I protein expression were higher than HG and HG + LV-Con group(P＜0.05),PLVAP,p-mTOR,mTOR mRNA,and PLVAP protein expression were lower than HG and HG+LV-Con group(P＜0.05).Comparison with HG + LV-CTRP13,p-mTOR protein expression in HG+LV-CTRP13+Compound C group increased(P＜0.05),while expressions of CTRP13,Beclin1 and LC3II/LC3I protein decreased(P＜0.05);the protein expressions of p-AMPK,Beclin1 and LC3II/LC3I were increased in HG+LV+ CTRP13+Torin1 group(P＜0.05),while CTRP13 and p-mTOR protein expression was decreased(P＜0.05);protein expressions of p-AMPK,p-mTOR and LC3II/LC3I were higher in HG+LV-CTRP13 + 3MA group(P＜0.05),while LC3II/LC3I protein expression was lower(P＜0.05).Conclusion CTRP13 overexpression activates AMPK/mTOR-autophagy signaling pathway,which may play a protective role in the function of rLSECs anddelay liver sinusoid capillarization.
