| OBJECTIVE: To investigate the effect of zinc and vitamin E in the expression of apoptosis associated genes Bcl-2 and Bax in placenta of clubfoot-like fetus induced by all-trans-retinoic acid(ATRA).METHODS: Pregnant Sprague-Dawley(SD) rats were randomly divided into three groups, including control group, experimental group and antiapoptotic intervention group. On the 10 th day of pregnancy, a dose of 120 mg/kg ATRA dissolved in mineral oil was given intragastrically to the rats in the experimental group and antiapoptotic intervention group, and equivalent volume of mineral oil was given intragastrically to the control rats. The rats in control group and experimental group fed with ordinary water and food. In the antiapoptotic intervention group, rats were fed with vitamin E(500 mg/kg) and Zn SO4?7H2O(227 mg/L) in deionized water. All pregnant rats were taken cesarean delivery operation on 20 th day of pregnancy, fetal rats and placentas were collected for the pathological and biochemical analysis.RESULTS: Clubfoot-like deformity fetal rats were observed in the experimental group and antiapoptotic intervention group, and all the fetal rats in control group were normal. Antiapoptotic intervention group had a lower Clubfoot-like deformity rate than the experimental group. Results of flow cytometry detection indicated that placental apoptosis rate in the experimental group higher than the control group, and the antiapoptotic intervention group was lower than the experimental group. Compared with the control group, both Real-time PCR and Western blotting all showed that lower expression of Bcl-2 and higher expression of Bax in the level of m RNA and protein in the experimental group. And compared with the experimental group, higher expression of Bcl-2 and lower expression of Bax in the level of m RNA and protein in the antiapoptotic intervention group. Results of immunohistochemical staining showed that positive expression rate of Bcl-2 is significantly reduced, while Bax protein expression is increased in the experimental group. And the opposite results in antiapoptotic intervention group.CONCLUSION: We found all-trans-retinoic acid has the effect of increasing apoptosis rate in pregnant rat placenta. Zinc and vitamin E can reduce the apoptosis effect in the placentas, thus to reducing the incidence of clubfoot-like fetal rats, which may through regulating the balance of Bcl-2/Bax. It can provide a new research direction for the further study of pathogenic mechanism and clinical prevention on congenital clubfoot. |
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