Department of Korean Medical Science, School of Korean Medicine, Pusan National University, Yangsan, Gyeongsangnam-do 50612, Republic of Korea; Korean Medicine Research Center for Healthy Aging, Pusan National University, Yangsan, Gyeongsangnam-do 50612, Republic of Korea.
Department of Korean Medical Science, School of Korean Medicine, Pusan National University, Yangsan, Gyeongsangnam-do 50612, Republic of Korea.
J Integr Med. 2021 Nov;19(6):537-544. doi: 10.1016/j.joim.2021.09.005. Epub 2021 Sep 14.
Mitophagy is known to contribute towards progression of Parkinson's disease. Korean red ginseng (KRG) is a widely used medicinal herb in East Asia, and recent studies have reported that KRG prevents 1-methyl-4-phenylpyridinium ion (MPP)-induced cell death. This study was undertaken to investigate whether KRG suppresses MPP-induced apoptosis and mitophagy.
SH-SY5Y cells were incubated with KRG for 24 h, and subsequently exposed to MPP. The MPP-induced cell death was confirmed with the 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, and the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay. Changes in the structure and function of mitochondria were confirmed using mitotracker, MitoSOX red mitochondrial superoxide indicator, parkin, and phosphatase and tensin homolog deleted on chromosome ten-induced putative kinase 1 (PINK1) immunofluorescent staining. Western blotting was performed to evaluate the expression of apoptosis-related factors in whole cells, including Bax, Bcl-2 and cleaved caspase-3, and mitophagy-related factors in the mitochondrial fraction, including cytochrome c, parkin, PINK1, translocase of the outer membrane 20 (TOM20), p62 and Beclin 1.
MPP induced cell death by cytochrome c release and caspase-3 activation; however, this effect was suppressed by KRG's regulation of the expressions of Bcl-2 and Bax. Moreover, MPP exposure increased the mitochondrial expressions of parkin, PINK1, Beclin 1 and p62, and decreased TOM20, cytochrome c and Bcl-2 expressions. These MPP-induced changes in the mitochondrial fraction were attenuated by treatment with KRG.
KRG effectively prevents MPP-induced SH-SY5Y cell death by regulating cytochrome c release from mitochondria and PINK1/parkin-mediated mitophagy, through regulation of the Bcl-2 family.
自噬被认为有助于帕金森病的进展。红参是东亚广泛使用的药用植物，最近的研究报道红参可预防 1-甲基-4-苯基吡啶离子（MPP）诱导的细胞死亡。本研究旨在探讨红参是否抑制 MPP 诱导的细胞凋亡和自噬。
将 SH-SY5Y 细胞与红参孵育 24 小时，然后暴露于 MPP 中。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐测定法和末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记法确认 MPP 诱导的细胞死亡。使用线粒体追踪剂、MitoSOX 红色线粒体超氧化物指示剂、parkin 和磷酸酶和张力蛋白同源物缺失的染色体 10 诱导的假定激酶 1（PINK1）免疫荧光染色来确认线粒体结构和功能的变化。进行 Western blot 以评估全细胞中凋亡相关因子（包括 Bax、Bcl-2 和 cleaved caspase-3）和线粒体部分中自噬相关因子（包括细胞色素 c、parkin、PINK1、外膜 20 转位酶（TOM20）、p62 和 Beclin 1）的表达。
MPP 通过细胞色素 c 释放和 caspase-3 激活诱导细胞死亡；然而，红参通过调节 Bcl-2 和 Bax 的表达抑制了这种作用。此外，MPP 暴露增加了线粒体 parkin、PINK1、Beclin 1 和 p62 的表达，降低了 TOM20、细胞色素 c 和 Bcl-2 的表达。这些线粒体部分的 MPP 诱导变化通过红参的治疗而减弱。
红参通过调节 Bcl-2 家族，有效防止 MPP 诱导的 SH-SY5Y 细胞死亡，从而防止线粒体细胞色素 c 释放和 PINK1/parkin 介导的自噬。
