| Perimenopausal syndrome cause by ovarian function decline.The decline of ovarian function is closely related to follicular atresia.Granulocytes are essential for follicular atresia.At home and abroad many studies have showed that hUCMSCs transplantation can improve ovarian function and reduce ovarian granulosa cell apoptosis.But the improving mechanism is not clear.In this study,we selected the postmenopausal rats of natural aging as models,then transplantated hUCMSCs to rats by tail vein.Calibrated the levels of Bcl-2,Bax and Caspase-3 which were detected in granulosa cells.To investigate whether hUCMSCs transplantation can reduce the expression of ovarian granulosa cells in peri-menopausal rats and provide the theoretical basis for the clinical application of hUCMSCs transplantation to delay female ovarian function decline.Objective1.The purpose of this study was to explore the hUCMSCs transplantation can improve the ovarian reserve function of perimenopausal rats by regulating the expression of apoptotic proteins in granulosa cells.2.The purpose of this study was to explore the hUCMSCs the expression of Bcl-2,Bax and Caspase-3 in granulosa cells at different time.MethodForty SPF level SD rats aged 12-14 months were selected,According to the vaginal smear of rat rats,Thirty perinatal rats whose vaginal smear disodered were screened as model.Rrandomly divided these rats into group A and group B(n = 15).Fifteen young female SD rats aged 3-5 months were selected as group C.All rats live and feed in a homogeneous manner.Group A was transplanted through tail vein graft cell buffer(PBS)1ml,every other day,a total of 2 times.Group B was transplanted with 1×106 cells /ml hUCMSCs single cell suspension by tail vein transplantation 1ml,every other day,a total of 2 times.Group C is not subject to any treatment.The expression of Bcl-2,Bax and Caspase-3 were detected by immunohistochemistry and western blot at 2 weeks,3weeks and 4 weeks after transplantation of hUCMSCs.Results1.Perimenopausal SD rats on2 groups are good in spirit,diet,sleep,coat color,activity and other.No bleeding and other immune rejection,no death occurred.2.Peripheral meningoceles ovarian granulosa cell apoptosis-related protein Bcl-2:(1)Western blot results: The expressions of Bcl-2 in group A and group B were significantly lower than that in group C at 2 weeks,3 weeks and 4 weeks after transplantation.2 weeks and 3 weeks after transplantation,the expressions of group A were less than that of group B,but there was no significant difference(P>0.05).But there was significant difference at 4 weeks after transplantation.The expression of group B at 2weeks was significantly lower than that at 4 weeks after transplantation(P <0.05).(2)Immunohistochemical results: The expressions of Bcl-2 in group A and group B were significantly lower than that in group C at 2 weeks,3 weeks and 4 weeks after transplantation.2 weeks and 3 weeks after transplantation,the expressions of group A were less than that of group B,but there was no significant difference(P>0.05).But there was significant difference at 4 weeks after transplantation.The expression of group B at 2weeks was significantly lower than that at 4 weeks after transplantation(P <0.05).3.Apoptosis-related protein of ovarian granulosa cells in peri-menopausal SD rats Bax:(1)Western blot results: The expressions of Bax in group A and group B were significantly higher than that in group C at 2 weeks,3 weeks and 4 weeks after transplantation(P < 0.05).2 weeks and 3 weeks after transplantation,the expression of group A was higher than that of group B,but there was no significant difference(P <0.05).But there were statistically differeces at 4 weeks(P <0.05).4 weeks after transplantationof group B was significantly lower than that in group 2(P <0.05).(2)Results of immunohistochemistry: The expressions of Bax in group A and group B were significantly higher than that in group C at 2 weeks,3 weeks and 4 weeks after transplantation(P < 0.05).2 weeks and 3 weeks after transplantation,the expression of group A was higher than that of group B,but there was no significant difference(P <0.05).But there were statistically differeces at 4 weeks(P <0.05).4 weeks after transplantationof group B was significantly lower than that in group 2(P <0.05).4.Bcl-2 / Bax ratio of ovarian granulosa cells in peri-menopausal SD rats:(1)Western blot results: The expressions of Bcl-2 in group A and group B were significantly lower than that in group C at 2 weeks,3 weeks and 4 weeks after transplantation.2 weeks and 3 weeks after transplantation,the expressions of group A were less than that of group B,but there was no significant difference(P>0.05).But there was significant difference at 4 weeks after transplantation.The expression of group B at 2weeks was significantly lower than that at 4 weeks after transplantation(P <0.05).(2)Results of immunohistochemistry: The expressions of Bcl-2 in group A and group B were significantly lower than that in group C at 2 weeks,3 weeks and 4 weeks after transplantation.2 weeksafter transplantation,the expressions of group A were less than that of group B,but there was no significant difference(P>0.05).But there were significant differences at 3 weeks and 4 weeks after transplantation.The expression of group B at 2 weeks was significantly lower than that at 4 weeks after transplantation(P<0.05).5.The ratio of Caspase-3 in ovarian granulosa cells of perimenopausal SD rats:(1)Western blot results: 2 weeks,3 weeks and 4 weeks after transplantation,theexpressions of group A and group B were significantly higherthan that of group C(P<0.05).The expression of group A was significantly higher than that of group B.There was no significant difference between the two groups at 2 weeksand 3 weeks(P> 0.05).But the results were statistically different at 4 weeks(P <0.05).The expression of 2 weeks in group B was significantly higher than that in 4 weeks after transplantation(P<0.05).(2)Immunohistochemical results: 2 weeks,3 weeks and 4 weeks after transplantation,theexpressions of group A and group B were significantly higherthan that of group C(P <0.05).The expression of group A was significantly higher than that of group B.There was no significant difference between the two groups at 2 weeksand3 weeks(P> 0.05).But the results were statistically different at 4 weeks(P <0.05).The expression of 2 weeks in group B was significantly higher than that in 4 weeks after transplantation(P <0.05).Conclusion1 The expression of Bcl-2,Bax and Caspase-3 in granulosa cells was reduced by transplantation of hUCMSCs.2 The regulation of apoptosis-related proteins in h FPMSCs at 4 weeks after transplantation was more significant at 2 weeks and 3 weeks after transplantation. |
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