Effect And Mechanism Of Kallistatin Gene On Thin Endometrium

OBJECTIVE: To investigate the role of KLST gene in thin endometrium,and to initially clarify the mechanism of KLST gene therapy for thin endometrium,so as to provide thinking and targeting for the therapy of thin endometrium.METHODS: Part I: Forty female SD rats were mean division into normal control Group,Sham Group,Model Group,Virus Empty Group(VE Group)and knockdown KLST Group(sh-KLST Group);HE staining,immunohistochemical staining and masson staining was used for testing the biological characteristics of the endometrium.Western blot was used to test the expression levels of KLST,PCNA,Bcl-2,casepase-3,Bax,MMP2,MMP9,PI3K/p-PI3 K and Akt/p-Akt.Mating experiment was used to detect the embryos of rats.Part II: Fifty female SD rats were evenly divided into Normal Control Group,Sham Group,Model Group,Virus Empty Group(VE Group),and Over Expression KLST Group(OE-KLST Group).HE staining,immunohistochemical staining and masson staining was used for testing the biological characteristics of the endometrium.Western blot was used to test the expression of KLST,PCNA,MMP2,MMP9,GSK-3β,VEGF,PI3K/p-PI3 K,Akt/p-Akt.Mating experiment was used to count embryos of rats.RESULTS: Compared with the normal control group,the endometrial thickness(P<0.01),expression of KLST,PCNA(P<0.001),and the number of pregnant embryos decreased(P<0.05)of sh-KLST group was decreased significantly.The area of intimal fibrosis and the expression of MMP2,MMP9 was significantly increased(P<0.001,P<0.001,P<0.01).The expression of Bax and casepase3 was increased(P<0.01),and the expression of Bcl-2 was decreased(P<0.01).The levels of pPI3K/PI3 K and p-Akt/Akt were significantly reduced(P<0.001).The thickness,KLST expression and PCNA expression in the endometrium of the model group were declined(P<0.01),the number of pregnant embryos was reduced(P<0.01),and the area of intimal fibrosis was elevated(P<0.001).The expressions of fibrosis-related proteins MMP2 and MMP9 were significantly raised(P<0.01),while the number of glands was significantly declined(P<0.01).The expressions of GSK-3β and VEGF were significantly decreased(P<0.001).After KLST overexpression gene therapy,the endometrial thickness,KLST expression and PCNA expression levels were increased(P<0.01),and the number of pregnant embryos was elevated(P<0.05).The area of fibrosis was reduced significantly(P<0.01),and the expression of MMP2 and MMP9 was significantly declined(P<0.001).The gland number was elevated(P<0.05),and the level of GSK-3β and VEGF were upward(P<0.01).The levels of p-PI3K/PI3 K and p-Akt/Akt were significantly elevated(P<0.001).Conclusion: Knockdown of KLST gene inhibits PI3K/Akt pathway,resulting in decreased cell proliferation,increased fibrosis and apoptosis in the endometrium of rats,which leads to a decrease in the number of pregnant embryos.Overexpression of KLST gene activates PI3K/Akt pathway and increases the number of pregnant embryos and pregnancy rate in rats by promoting endometrial proliferation,reducing endometrial fibrosis and angiogenesis.KLST gene is expected to become a new target for the treatment of thin endometrium.