- Author:
Zhi-Yu DUAN
1
;
Xiao-Rui WANG
1
;
Qing-Shan LI
1
;
Tai-Gang LIANG
1
;
Ting-Ting LIANG
2
;
Qing-Shan LI
3
Author Information
- Publication Type:Journal Article
- Keywords: apoptosis; dibutyltin; HL02; mitochondria; ROS; sinomenine
- From: Chinese Pharmacological Bulletin 2022;38(10):1535-1541
- CountryChina
- Language:Chinese
- Abstract: Aim To investigate the protective effect of sinomenine(SIN)against dibutyltin(DBT)induced injury in HL02 cells and explore the potential mechanism.Methods HL02 cells were cultured and divided into control,model and SIN-treated groups.Cell proliferation was detected by MTT method.Cell morphology was observed.Cell apoptosis was detected by Acridine orange/ethidium bromide(AO/EB)fluorescent staining and Annexin V-FITC/PI double staining.Meanwhile,intracellular reactive oxygen species(ROS)concentration was detected by DCFH-DA staining.Mitochondrial membrane potential(MMP)was tested by JC-1 dye.Moreover,the mRNA expression of apoptosis-related proteins was detected by qRT-PCR,and the protein expression of Bcl-2,Bax,caspase-9,cleaved-caspase-3 were measured via Western blot.Results The pretreatment with SIN increased the cell viability and decreased morphological changes induced by DBT in a dose-dependent manner.Meanwhile,cell apoptotic rates and intracellular ROS decreased,and the loss of MMP was partially restored.Compared to DBT-treated group,SIN treatment could increase the mRNA levels of Bcl-2,Bcl-xL and decrease the mRNA levels of Bax,Bad,cytochrome-c,Apaf-1,caspase-9 and caspase-3.Furthermore,SIN could significantly up-regulated the DBT-induced decrease in Bcl-2/Bax ratio,and down-regulated the DBT-induced over-expressions of caspase-9 and cleaved-caspase-3.Conclusions SIN could protect HL02 cells against DBT-induced cell injury,which is related to the inhibition of ROS-mediated mitochondrial apoptosis.