The Protection Effects Of Cyanidin-3-O-Glucoside On Progesterone Biosynthesis Against Pb In R2C Rat Leydig Cells

Objective:The protection effect of Cyanidin-3-O-glucoside?C3G? on progesterone biosynthesis against lead in R2 C rat leydig cells was studied.Method:Cells were treated with Pb²⁺ at various concentrations for 24 h, and progesterone concentration in the medium was analyzed using radioimmunoassay?RIA? to select the Pb concentration for the following experiments. Next, cell were treated with Pb?at the selected concentration? with or without C3 G, and cell viability was measured using CCK8 kit, cell morphology was observed using microscope, progesterone concentration in the medium was analyzed using RIA, Reactive oxygen species?ROS? level was detected using chemiluminescence?CL?, the ratio of the number of cells with low mitochondrial membrane potential?MMP? to the number of total cells was determined using flow cytometry?FCM?, and the expression of proteins related to progesterone biosynthesis or oxidation resistance were analyzed using Western Blot.Results:100 ?M Pb was used to induce the decrease of progesterone biosynthesis, and 12.5, 25, 50, 100 ?M C3 G were used to alleviate Pb-induced damages. After being treated with Pb and /or C3 G for 48 h, cell vability in Pb treated groups significantly decreased compared with control and that of Pb and 12.5, 25 or 50 ?M C3 G treated groups significantly increased compared with Pb treated groups. After being treated with Pb and /or C3 G 24h, there is no significant difference in cell viability among all the groups, but the progesterone concentration in Pb treated groups decreased significantly compared with control, 12.5, 25 or 50 ?M C3 G increased the progesterone concentration. The expression of proteins St AR, 3?-HSD, CYP11A1, PKA and p-ERK1/2 / REK1/2 ratio decreased significantly in Pb treated groups?24h? compared with control, and that of Pb and 25 ?M C3 G treated group significantly increased compared with Pb treated groups?24h?. The expression of proteins in SOD2 and CAT significantly decreased in Pb treated group?24h? compared with control, and that of 25 and 12.5?M C3 G alleviated the decrease of SOD2 and CAT, respectively?24h?. The MMP damage ratio of cells increased significantly after being treated with Pb for 24 h, and MMP in all the C3 G + Pb treated groups were found to be significantly lower than that of Pb treated groups. Pb treatment for 0-160 min significantly increased the ROS generation, and ROS level decreased significantly in all Pb+C3G treated groups compared with Pb treated groups. The ROS level of 12.5 ?M C3 G + Pb treated groups turned out to be the closest to that of control.Conclusion:Pb decreased the progesterone biosynthesis in R2 C rat leydig cells by inhibiting the PKA-ERK1/2 pathway, increasing the oxidative stress and down-regulating the expression of St AR?CYP11A1 and 3?-HSD. C3 G alleviated the Pb-induced decrease of progesterone biosynthesis by activating the PKA-ERK1/2 pathway, decreasing the oxidative stress and up-regulating the expression of St AR?CYP11A1 and 3?-HSD...