Vitamin C Regulates Bovine Cumulus Cell Proliferation And Apoptosis Through COL1A1

Vitamin C(Vitamin C,VC),also known as L-ascorbic acid,is not only an important substance for maintaining life activities in the body,but also plays an important role in many cellular processes in the body.Studies have shown that vitamin C can not only effectively remove reactive oxygen free radicals(ROS)in cells,but also affect the development of bovine ovary by regulating the physiological state of cumulus cells,especially the level of apoptosis and autophagy,but the specific approach and mechanism are not clear.In mammalian ovaries,the extracellular matrix(ECM)has important effects on follicular development and ovarian function.Each type of ECM collagen is formed by three identical or different peptide chains,forming a triple helix.Among them,the type I collagen has the most typical structure.Type I collagen ?1 chain(COL1A1),as the main component of extracellular matrix,plays an important role in regulating the physiological state of some cancer cells.However,its role in bovine cumulus cells remains unclear.In order to investigate the effect of vitamin C and structural proteins of ECM on cumulus cells,we treated bovine cumulus cells with vitamin C and explored its effects on the physiological state and function of cumulus cells through COL1A1.Therefore,in this study,cumulus cells were first cultured in vitro,the effect of vitamin C on apoptosis was investigated by flow cytometry,and its effect on cell proliferation was explored by CCK-8.In addition,the effect of q RT-PCR on the expression of genes related to extracellular matrix structure.The results show that 100 ?M of vitamin C can effectively inhibit bovine cumulus cell apoptosis,and this concentration of vitamin C can significantly promote cell replication levels.The results of q RT-PCR showed that the addition of vitamin C could increase the expression levels of COL1A1 and COL1A2 genes,which had a significant effect on COL1A1.Subsequently,in order to clarify the specific role of COL1A1,this study inhibited the expression of COL1A1 in cumulus cells by si RNA,and determined the cumulus cell cycle and proliferation levels by flow cytometry and CCK-8 assay,respectively.The mitochondrial membrane potential changes were measured by JC-1.Detection of cell autophagy level by immunofluorescence;detection of cell migration and diffusion ability by cell scratch assay and q RT-PCR;detection of lactic acid and cholesterol concentration by a microplate reader to evaluate glucose utilization and cholesterol synthesis activity in cumulus cells;q RT-PCR and western-blot analysis are used to measure changes in key gene expression.The results showed that after COL1A1 expression was knocked down by si RNA,bovine cumulus cells had an abnormal cell cycle,the number of S-phase cells was significantly reduced,cholesterol synthesis was reduced,cell proliferation ability was decreased,and apoptosis rate was increased.These findings may be due to the inhibition of COL1A1 resulting in high ROS content in cells,decreased mitochondrial membrane potential,increased intracellular autophagy,activation of apoptotic pathways,and decreased lactic acid conversion capacity.It is shown that COL1A1 plays an important role in regulating the physiological and biological functions of bovine cumulus cells.In summary,we found that in addition to affecting cell apoptosis and proliferation,vitamin C can also affect the expression of the structural protein COL1A1 of the extracellular matrix,and thus play a potential role in various important physiological functions of bovine cumulus cells.This study provides new ideas for further understanding the biological functions of vitamin C and the effective use of vitamin C,and provides a theoretical basis for the mechanism of the effect of extracellular matrix on the apoptosis and proliferation of bovine cumulus cells.