人脐带间充质干细胞来源的外泌体通过Nrf2/NQO-1途径改善实验性非酒精性脂肪性肝炎。
Exosomes derived from human umbilical cord mesenchymal stem cells ameliorate experimental non-alcoholic steatohepatitis via Nrf2/NQO-1 pathway.
作者信息
Kang Yaxing, Song Yiran, Luo Yuxin, Song Jia, Li Chenyang, Yang Shuangshuang, Guo Jinbo, Yu Jun, Zhang Xiaolan
机构信息
Department of Gastroenterology, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, China; Department of Endocrinology, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
Department of Gastroenterology, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
出版信息
Free Radic Biol Med. 2022 Nov 1;192:25-36. doi: 10.1016/j.freeradbiomed.2022.08.037. Epub 2022 Sep 10.
BACKGROUND
No approved effective therapy for non-alcoholic steatohepatitis (NASH) is currently available. Exosomes derived from mesenchymal stem cells (MSCs) perform the functions such as inhibiting inflammation, anti-oxidative stress, regulating immunity, but it is not clear whether human umbilical cord mesenchymal stem cells (hUC-MSCs) exosomes protect against NASH through Nrf2/NQO-1 pathway. Therefore, this study was conducted to investigate the effects of hUC-MSCs exosomes on NASH through Nrf2/NQO-1 pathway in vivo and in vitro.
METHODS
C57BL/6J male mice were fed with high fat and high cholesterol diet (HFHC) and methionine choline deficiency diet (MCD). Mice were treated with or without hUC-MSCs exosomes by tail intravenous injection. The liver histology, lipid metabolism and oxidative stress were evaluated. HepG2 and AML12 cells were incubated with palmitic acid (PA) and MCD conditioned medium, respectively. Then the therapeutic effect of hUC-MSCs exosomes in steatotic cells was evaluated. To elucidate the signaling pathways, the Nrf2-specific blocker ML385 was applied to intervene in vitro.
RESULTS
In NASH models, hUC-MSCs exosomes attenuated steatosis in hepatocytes, altered the abnormal expression of lipid-related genes including SREBP-1c, PPAR-α, Fabp5, CPT1α, ACOX and FAS, suppressed the hepatic inflammatory responses by decreasing the expression of F4/80 macrophages, CD11c macrophages as well as the content of TNF-α and IL-6. hUC-MSCs exosomes also inhibited oxidative stress by reducing the level of MDA, CYP2E1 and ROS, increasing the activity of SOD and GSH in hepatocytes. Notably, hUC-MSCs exosomes enhanced the protein ratio of p-Nrf2/Nrf2 and the protein expression of NQO-1. Moreover, in vitro, the therapeutic effects of hUC-MSCs exosomes on lipid deposition and ROS were reversed by ML385. Also, ML385 reduced the protein expression of p-Nrf2 and NQO-1 in vitro.
CONCLUSION
Nrf2/NQO-1 antioxidant signaling pathway may play a key role in the treatment of NASH by hUC-MSCs exosomes.
背景
目前尚无经批准的非酒精性脂肪性肝炎（NASH）有效治疗方法。间充质干细胞（MSCs）来源的外泌体具有抑制炎症、抗氧化应激、调节免疫等功能，但人脐带间充质干细胞（hUC-MSCs）外泌体是否通过Nrf2/NQO-1途径预防NASH尚不清楚。因此，本研究旨在通过体内和体外实验研究hUC-MSCs外泌体通过Nrf2/NQO-1途径对NASH的影响。
方法
将C57BL/6J雄性小鼠喂以高脂肪高胆固醇饮食（HFHC）和蛋氨酸胆碱缺乏饮食（MCD）。通过尾静脉注射对小鼠进行有无hUC-MSCs外泌体的处理。评估肝脏组织学、脂质代谢和氧化应激情况。分别用棕榈酸（PA）和MCD条件培养基培养HepG2和AML12细胞。然后评估hUC-MSCs外泌体对脂肪变性细胞的治疗效果。为阐明信号通路，应用Nrf2特异性阻滞剂ML385进行体外干预。
结果
在NASH模型中，hUC-MSCs外泌体减轻了肝细胞脂肪变性，改变了包括SREBP-1c、PPAR-α、Fabp5、CPT1α、ACOX和FAS在内的脂质相关基因的异常表达，通过降低F4/80巨噬细胞、CD11c巨噬细胞的表达以及TNF-α和IL-6的含量抑制了肝脏炎症反应。hUC-MSCs外泌体还通过降低MDA、CYP2E1和ROS水平，增加肝细胞中SOD和GSH的活性来抑制氧化应激。值得注意的是，hUC-MSCs外泌体提高了p-Nrf2/Nrf2的蛋白比例和NQO-1的蛋白表达。此外，在体外，ML385逆转了hUC-MSCs外泌体对脂质沉积和ROS的治疗效果。ML385还降低了体外p-Nrf2和NQO-1的蛋白表达。
结论
Nrf2/NQO-1抗氧化信号通路可能在hUC-MSCs外泌体治疗NASH中起关键作用。
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