Effect of Ranae Oviductus Protein Hydrolysate on Ethanol-induced L-02 Cell Injury
10.13422/j.cnki.syfjx.20211404
- VernacularTitle:哈蟆油蛋白酶解物对乙醇诱导的L-02细胞损伤的作用
- Author:
Yi ZHANG
1
;
Ran ZHENG
1
;
Qi YU
1
;
Li-li JIAO
1
;
Bo LI
1
;
Da LIU
1
;
Yi-ping LI
1
Author Information
1. Changchun University of Chinese Medicine，Changchun 130117，China
- Publication Type:Research Article
- Keywords:
Ranae Oviductus protein hydrolysate （ROPH）;
alcoholic liver cell injury;
oxidative stress;
mitochondrial apoptosis;
inflammatory response
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(15):43-50
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the protective effect and mechanism of Ranae Oviductus protein hydrolysate （ROPH） on the expression of pathway-related proteins in ethanol-induced L-02 cell injury. Method:The ROPH was prepared by compound enzymatic hydrolysis. L-02 cell injury model was induced with 400 mmol·L-1 ethanol. Cell viability was detected by cell counting kit-8 （CCK-8） assay. Cell cycle and apoptosis were examined by flow cytometry. JC-1/Hochest staining was employed for qualitative investigation. The expression of related proteins in apoptosis， mitogen-activated protein kinase （MAPK） signaling pathway， and pyroptosis in L-02 cells was detected by Western blot. Result:The results of the CCK-8 assay showed that 400 mmol·L-1 ethanol could induce L-02 cell injury within 12 hours. Compared with the blank group， the model group showed decreased viability of L-02 cells （P<0.01）， elevated percentage of the cell cycle in the G0/G1 phase （P<0.01）， increased total cell apoptosis rate （P<0.01）， reduced mitochondrial membrane potential （P<0.01）， up-regulated expression of apoptosis-related proteins ［B-cell lymphoma-2 （Bcl-2）-associated X protein （Bax）， Cytochrome C （Cyt C）， and cysteine-dependent aspartate specific protease-3 （Caspase-3）］ （P<0.05， P<0.01） and MAPK signaling pathway-related proteins ［C-Jun amino-terminal kinase （JNK） and p38 MAPK］ （P<0.05， P<0.01）， and potentiated expression of pyrolysis-related proteins Caspase-1 and interleukin-1β （IL-1β） （P<0.05）. Compared with the model group， the ROPH treatment group exhibited improved cell cycle arrest （P<0.05， P<0.01）， diminished total cell apoptosis rate （P<0.01）， elevated mitochondrial membrane potential in a dose-dependent manner， down-regulated expression of Bax， Cyt C， and Caspase-3 proteins （P<0.05， P<0.01）， up-regulated expression of Bcl-2 protein （P<0.05， P<0.01）， and a downward trend in expression of proteins related to MAPK signaling pathway and pyrolysis （P<0.05， P<0.01）. Conclusion:ROPH could inhibit oxidative stress-triggered liver injury in ethanol-induced cells by improving mitochondrial membrane potential， reducing the expression of proteins in the mitochondria-mediated apoptosis pathway， and inhibiting the expression of proteins related to the MAPK signaling pathway and pyrolysis pathway to reduce the mitochondrial dysfunction and inflammatory response in ethanol-induced L-02 liver cells and inhibit oxidative stress， thereby exerting a therapeutic role in alcoholic liver injury.
