| The stress response have cause physical and psychological harm to female animals.Stress reaction will affect the development of oocytes.Mural granulosa cells（MGCs）and Oviductal epithelial cells（OECs）are closely related to the development of oocytes.In mice,binding stress can activate the hypothalamic-pituitary-adrenal（HPA）axis,promote thesecretion of stress-related hormone Corticosterone（CORT）,and induce the apoptosis ofMGCs and OECs.Tissue type plasminogen activator（tPA）is related to cell apoptosis,but the role and mechanism of tPA in Corticosterone-induced apoptosis of MGCs and OECs areunclear.In our vitro experiment,Corticosterone was added to induce apoptosis of MGCs and OECs,and the expression of tPA encoding gene PLAT and the effect of tPA on apoptosis of MGCs and OECs were determined.The expression of LRP-1 and ANX-A2 was knockeddown by RNAi technique in vitro to determine the pathway of the effect of tPA on apoptosis.By studying the effect of tPA on the apoptosis of MGCs and OECs induced by Corticosterone in mice,it is expected to provide theoretical and technical support for the study of themechanism of stress and Corticosterone-induced oocyte development,the application of animal embryo engineering technology for the proliferation and assisted reproduction oflaboratory animals and domestic animals,and the research and development of target drugs related to female reproductive function.The results are as follows:1.Compared with the control group,the proportion of healthy cells decreasedsignificantly and the proportion of apoptosis increased significantly（healthy cells:86.0%vs74.5%,early apoptosis:6.7%vs 12.3%）after the addition of CORT(concentration of 1×10-5M)in vitro cultured granulosa cells（P<0.05）.Compared with the control group,thepercentage of healthy cells decreased and the percentage of apoptotic cells increasedsignificantly（healthy cells:86.2%vs 78.8%,early apoptotic cells:8.2%vs 14.6%）in vitrocultured tubal epithelial cells supplemented with CORT(concentration of 1×10-5 M)（P<0.05）.2.The expression of PLAT of tPA gene in granulocyte cells after CORT treatment was decreased（0.76±0.05）,and the difference was significant（P<0.05）,the expression of PLAT gene of tPA in oviduct epithelial cells after CORT treatment was decreased（0.77±0.07）,and the difference was significant（P<0.05）.3.On the premise of CORT-induced apoptosis of MGCs,CORT and tPA were addedtogether to treat cells.The results showed that 1×10-8 M and 2×10-8 M tPA had no significant effect on MGCs,while the ratio of Bcl-2/Bax was significantly increased after 4×10-8 M tPA and CORT combined treatment compared with the control group（P<0.05）,the proportion of healthy cells was significantly increased,and the proportion of early apoptosis wassignificantly decreased（healthy cells:74.5%vs 81.9%,early apoptosis:12.3%vs 8.7%）（P<0.05）.Under the premise of CORT-induced apoptosis of OECs,the combined addition of CORT and TPA showed no significant effect on OECs,while the combined treatment of 4×10-8M TPA and CORT significantly increased the Bcl-2/Bax ratio compared with the controlgroup（P<0.05）,the proportion of healthy cells was significantly increased,and the proportion of early apoptosis was significantly decreased（healthy cells:78.8%vs 85.1%,earlyapoptosis:14.6%vs 9.2%）（P<0.05）.4.In MGCs,the Bcl-2/Bax ratio in the LRP-1-si RNA interference group wassignificantly decreased compared with the NC-si RNA interference negative control group（P<0.05）,apoptosis.5.In OECs,the Bcl-2/Bax ratio in the LRP-1-si RNA interference group was significantly increased compared with the NC-si RNA interference negative control group（P<0.05）without apoptosis.6.In OECs,the Bcl-2/Bax ratio in the ANX-A2-si RNA interference group wassignificantly decreased（P<0.05）compared with the NC-si RNA interference negative control group,resulting in apoptosis.In conclusion,the addition of CORT can promote the apoptosis of MGCs and OECs.In MGCs,tPA plays an anti-apoptotic role through the LRP-1 pathway.In OECs,tPA exerts an anti-apoptotic effect through the ANX-A2 pathway.tPA plays an anti-apoptotic role in cort-induced apoptosis of mouse oviduct epithelial cells and granulosa cells of ovarian wall.It is of great significance to improve the dysplasia of oocytes caused by psychological stress. |
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