Study on the mechanism of Wuzi-Yanzong-Wan-medicated serum interfering with the mitochondrial permeability transition pore in the GC-2 cell induced by atractyloside.
10.1016/S1875-5364(22)60153-5
- Author:
De-Ling WU
1
,
2
,
3
;
Tong-Sheng WANG
4
;
Hong-Juan LIU
4
;
Wei ZHANG
4
;
Xiao-Hui TONG
4
;
Dai-Yin PENG
5
;
Ling-Yi KONG
6
Author Information
1. State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Bioactive Natural Product Research, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 210009, China
2. School of Pharmacy, Anhui University of Chinese Medicine, Anhui Province Key Laboratory of Chinese Medical Formula, Hefei 230012, China
3. School of Integrated Traditional Chinese and Western Medicine, Anhui University of Chinese Medicine, Hefei 230012, China.
4. School of Pharmacy, Anhui University of Chinese Medicine, Anhui Province Key Laboratory of Chinese Medical Formula, Hefei 230012, China.
5. School of Pharmacy, Anhui University of Chinese Medicine, Anhui Province Key Laboratory of Chinese Medical Formula, Hefei 230012, China. Electronic address: pengdy@ahtcm.edu.cn.
6. State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Bioactive Natural Product Research, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 210009, China. Electronic address: cpu_lykong@126.com.
- Publication Type:Journal Article
- Keywords:
Cyclophilin D;
Mitochondrial membrane potential;
Mitochondrial permeability transition pore;
Voltage-dependent anion channel 1;
Wuzi-Yanzong-Wan
- MeSH:
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine;
Animals;
Atractyloside/pharmacology*;
Cyclophilin D;
Male;
Matrix Metalloproteinases;
Mice;
Mitochondrial Membrane Transport Proteins/metabolism*;
Mitochondrial Permeability Transition Pore;
RNA, Messenger;
Rats
- From:
Chinese Journal of Natural Medicines (English Ed.)
2022;20(4):282-289
- CountryChina
- Language:English
-
Abstract:
Wuzi-Yanzong-Wan (WZYZW) is a classic prescription for male infertility. Our previous investigation has demonstrated that it can inhibit sperm apoptosis via affecting mitochondria, but the underlying mechanisms are unclear. The purpose of the present study was to explore the actions of WZYZW on mitochondrial permeability transition pore (mPTP) in mouse spermatocyte cell line (GC-2 cells) opened by atractyloside (ATR). At first, WZYZW-medicated serum was prepared from rats following oral administration of WZYZW for 7 days. GC-2 cells were divided into control group, model group, positive group, as well as 5%, 10%, 15% WZYZW-medicated serum group. Cyclosporine A (CsA) was used as a positive control. 50 μmol·L-1 ATR was added after drugs incubation. Cell viability was assessed using CCK-8. Apoptosis was detected using flow cytometry and TUNEL method. The opening of mPTP and mitochondrial membrane potential (MMP) were detected by Calcein AM and JC-1 fluorescent probe respectively. The mRNA and protein levels of voltage-dependent anion channel 1 (VDAC1), cyclophilin D (CypD), adenine nucleotide translocator (ANT), cytochrome C (Cyt C), caspase 3, 9 were detected by RT-PCR (real time quantity PCR) and Western blotting respectively. The results demonstrated that mPTP of GC-2 cells was opened after 24 hours of ATR treatment, resulting in decreased MMP and increased apoptosis. Pre-protection with WZYZ-medicated serum and CsA inhibited the opening of mPTP of GC-2 cells induced by ATR associated with increased MMP and decreased apoptosis. Moreover, the results of RT-qPCR and WB suggested that WZYZW-medicated serum could significantly reduce the mRNA and protein levels of VDAC1 and CypD, Caspase-3, 9 and CytC, as well as a increased ratio of Bcl/Bax. However, ANT was not significantly affected. Therefore, these findings indicated that WZYZW inhibited mitochondrial mediated apoptosis by attenuating the opening of mPTP in GC-2 cells. WZYZW-medicated serum inhibited the expressions of VDAC1 and CypD and increased the expression of Bcl-2, which affected the opening of mPTP and exerted protective and anti-apoptotic effects on GC-2 cell induced by ATR.
