Effect Of Muniziqi On The Expression Of PFN1 And CFL1 In Rats With Chronic Stress-induced Premature Ovarian Failure

Objective:This study was performed by replicating a rat model of chronic stress-induced premature ovarian failure and treating it with the drug.To investigate some of the differential proteins in chronic stress-induced ovarian premature aging rats based on iTRAQ combined with LC/MS/MS technique in previous work,and to verify the pathogenesis of chronic premature ovarian failure rats.Provide meaningful experimental data.Methods:A total of 90 SD rats with normal cycles were randomly selected and 10 randomly selected from the normal group(N group).The remaining 80 rats were set as chronic stress-type modules(CS group).Wet and cold,and kept in an environment where the day and night are upside down.Regularly observe the general condition of each group of rats(hair smoothness,exercise,agility,body weight,water intake,food intake,etc.)and the estrous cycle of each rat.Changes,by observing the general situation,rat estrus cycle and qualitative indicators(hormone FSH,E2,LH levels)to confirm the establishment of chronic stress model,confirmed from the chronic stress model to screen out the ovarian premature aging Rats were divided into ovarian premature aging group(POF group),ovarian premature aging drug treatment low dose group(PDL group),ovarian premature aging drug treatment middle dose group(PDM group),ovarian premature aging drug treatment high dose group(PDH group)(1)The levels of β-EP,FSH,E2 and LH in peripheral blood were detected by ELISA;(2)Morphological changes of ovarian tissue:(3)Screening by ELISA Differentially expressed egg The content of white PFN1 and CFL1 in the serum of each group;the expression of differentially expressed proteins PFN1 and CFL1 in the ovary were detected by immunohistochemistry;the expression of PFN1 and CFL1 mRNA in ovary was detected by RT-PCR.the amount.Results(1)General observation:During the modeling period,the rats in the chronic stress group had rough skin,loss of luster,markedly reduced exercise,and did not respond quickly,and fatigue was observed.The increase in weight and water intake of rats was significantly lower than that in normal rats;the increase in food intake;the general condition and estrous cycle of rats during drug treatment The disordered situation is effectively improved.(2)The E2 content in the peripheral serum of the premature ovarian failure group(POF group)and the FSH and LH levels were significantly higher than those in the normal group((P<0.05).The dose of drug treatment can regulate the hormone content in the serum of each group of rats.The serum β-EP content in the premature ovarian failure group and the high-dose group of ovarian premature aging drugs could significantly increase the serum p-EP content(P<0.05).(3)Morphological changes of ovarian tissue:no obvious ovarian follicles,vacuolar-like changes and severe edema in the ovarian premature aging group,and the improvement with the increase of drug dose also increased,among which the high-dose drug treatment group improved most,it is good.(4)Immunohistochemistry results showed that the expression of differential PFN1 protein in ovarian cytoplasm was higher than that in ovarian nucleus,but not in interstitial and non-expression or low expression in luteal,PFN1 protein in ovarian premature aging group(POF group)The expression of rat was significantly lower than that of N group(P<0.05).The expression of CFL1 protein in rat ovarian cytoplasm was increased compared with that in ovary nucleus.The expression of CFL1 protein in the premature ovarian failure group(POF group)was significantly higher than that in the N group(P<0.05),and the drug treatment group could reduce its expression.(5)Real-time quantitative RT-PCR CFL1 mRNA increased in the ovarian premature aging group(POF group)compared with the group(N group)(P<0.05).The high-dose drug treatment group in the group significantly down-regulated the mRNA expression levels of PFN1 and CFL1(P<0.05).Conclusion:Chronic stress can lead to premature ovarian failure;histopathological changes of ovary and hormonal disorders(E2,FSH,LH)may be one of the important mechanisms of its pathogenesis;the drug muscarin can improve the ovary In premature aging lesions,the pharmacological mechanism may be related to the regulation of disordered hormone levels(E2,FSH,LH)and the up-regulation of related proteins PFN1 and CFL1.The remaining 80 rats were set as chronic stress-type modules(CS group).Wet and cold,and kept in an environment where the day and night are upside down.Regularly observe the general condition of each group of rats(hair smoothness,exercise,agility,body weight,water intake,food intake,etc.)and the estrous cycle of each rat.Changes,by observing the general situation,rat estrus cycle and qualitative indicators(hormone FSH,E2,LH levels)to confirm the establishment of chronic stress model,confirmed from the chronic stress model to screen out the ovarian premature aging Rats were divided into ovarian premature aging group(POF group),ovarian premature aging drug treatment low dose group(PDL group),ovarian premature aging drug treatment middle dose group(PDM group),ovarian premature aging drug treatment high dose group(PDH group)(1)The levels of β-EP,FSH,E2 and LH in peripheral blood were detected by ELISA;(2)Morphological changes of ovarian tissue:(3)Screening by ELISA Differentially expressed egg The content of white PFN1 and CFL1 in the serum of each group;the expression of differentially expressed proteins PFN1 and CFL1 in the ovary were detected by immunohistochemistry;the expression of PFN1 and CFL1 mRNA in ovary was detected by RT-PCR.the amount.General observation:During the modeling period,the rats in the chronic stress group had rough skin,loss of luster,markedly reduced exercise,and did not respond quickly,and fatigue was observed.The increase in weight and water intake of rats was significantly lower than that in normal rats;the increase in food intake was significantly higher than that in normal rats(P<0.05);the general condition and estrous cycle of rats during drug treatment The disordered situation is effectively improved.The E2 content in the peripheral serum of the premature ovarian failure group(POF group)was significantly lower than that in the normal group(P<0.05),and the FSH and LH levels were significantly higher than those in the normal group(P<0.05).The dose of drug treatment can regulate the hormone content in the serum of each group of rats.The serum p-EP content in the premature ovarian failure group was significantly lower than that in the normal group(P<0.05),and the high-dose group of ovarian premature aging drugs could significantly increase the serum p-EP content(P<0.05).(3)Morphological changes of ovarian tissue:no obvious ovarian follicles,vacuolar-like changes and severe edema in the ovarian premature aging group,and the improvement with the increase of drug dose also increased,among which the high-dose drug treatment group improved most,it is good.(4)Immunohistochemistry results showed that the expression of differential PFN1 protein in ovarian cytoplasm was higher than that in ovarian nucleus,but not in interstitial and non-expression or low expression in luteal,PFN1 protein in ovarian premature aging group(POF group)The expression of rat was significantly lower than that of N group(P<0.05).The expression of CFL1 protein in rat ovarian cytoplasm was increased compared with that in ovary nucleus.The expression of CFL1 protein in the premature ovarian failure group(POF group)was significantly higher than that in the N group(P<0.05),and the drug treatment group could reduce its expression.(5)Real-time quantitative RT-PCR CFL1 mRNA increased in the ovarian premature aging group(POF group)compared with the group(N group)(P<0.05).The high-dose drug treatment group in the group significantly down-regulated the mRNA expression levels of PFN1 and CFL1(P<0.05).Conclusions:Chronic stress can lead to premature ovarian failure;histopathological changes of ovary and hormonal disorders(E2,pathogenesis;the drug muscarin can improve the ovary In premature aging lesions,This study was performed by replicating a rat model of chronic stress-induced premature ovarian failure and treating it with the drug.To investigate some of the differential proteins in chronic stress-induced ovarian premature aging rats based on iTRAQ combined with LC/MS/MS technique in previous work,and to verify the pathogenesis of chronic premature ovarian failure rats.Provide meaningful experimental data.METHODS:A total of 90 SD rats with normal cycles were randomly selected and 10 randomly selected from the normal group(N group).The remaining 80 rats were set as chronic stress-type modules(CS group).Wet and cold,and kept in an environment where the day and night are upside down.Regularly observe the general condition of each group of rats(hair smoothness,exercise,agility,body weight,water intake,food intake,etc.)and the estrous cycle of each rat.Changes,by observing the general situation,rat estrus cycle.