The purpose of this study was to investigate the effect of lysophosphatidic acid(LPA)with different concentrations on the expression of hyaluronate synthase 2(HAS2),prostaglandin-endoperoxide synthase 2(PTGS2)and pentraxin 3(PTX3)in Yak cumulus cells(YCCs).The YCCs of healthy adult(3-4 years old)female yaks were taken as the research object.YCCs in logarithmic growth phase were taken,and different concentrations of LPA(blank control,nega-tive control,5,15,30,and 50 μmol·L-1)were applied to the cultured YCCs in vitro.After 12,24,36,and 48 hours of cultivation,CCK-8 was used to detect the cellular viability of YCCs.The relative expression levels of HAS2,PTGS2,and PTX3 mRNA and protein in YCCs were detec-ted by RT-qPCR and Western-blot assays.Immunofluorescence staining was used to detect the distribution of cumulus dilatation factors HAS2,PTGS2,and PTX3 in YCCs.In this experi-ment,each treatment group had 3 replicates.The results showed that,LPA incubation for 12,24,36,and 48 h had a significant promoting effect on the viability of YCCs.When the incubation time was 24 h,LPA had the most significant promoting effect on the viability of YCCs.Com-pared to the control group,when the LPA concentration was 15 μmol·L-1,the viability of YCCs increased most significantly at different incubation times(P＜0.05).Compared with the blank control group,the highest relative expression levels of HAS2,PTGS2,and PTX3 mRNA and protein were observed(P＜0.05)when the LPA concentration was 15 μmol·L-1,and the fluo-rescence intensity of HAS2,PTGS2,and PTX3 in YCCs was significantly enhanced.However,when the LPA concentration exceeded 15 μmol·L-1,the relative expression levels of HAS2,PTGS2,and PTX3 mRNA and protein gradually decreased.The study results suggests that LPA has a promoting effect on the viability of YCCs.When the incubation time is 24 hours and the LPA concentration is 15 μmol·L-1,the viability improvement is the most significant(P＜0.05).LPA can enhance the expression of the cumulus expansion factors HAS2,PTGS2,and PTX3 in YCCs,and its effect is dose-dependent,with the optimal concentration being 15 μmol·L-1.The research results provide a theoretical basis for elucidating the molecular mechanism of LPA pro-moting the expansion of yak cumulus cells,and for further improving the quality of yak oocytes and the success rate of in vitro fertilization(IVF).
维普
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