Expression Levels Of Progesterone And Its Synthesis Related Factors In Rana Oil

As a precious economic animal in Northeast China,the oviduct of the Chinese brown frog(Rana dybowskii)goes expansion specifically during pre-hibernation to form Rana frog oil.As a functional food,Rana oil is rich in sex steroid hormones and has the function of regulating human endocrine function.However,it is unclear whether Rana oil can synthesize steroid hormones de novo.This project takes Rana oil-the oviduct of Chinese brown frog in pre-hibernation as research objects,and the oviduct obtained in the breeding period was used as a control.The expression levels of cholesterol,progesterone,and progesterone synthesis-related factors in the oviduct are measured to provide important scientific theoretical basis of the synthesis pathway of progesterone and the synthetic pathway of sex steroid hormones in the oviduct of Chinese brown frog.At the same time,the research results also lay the foundation for the sustainable and in-depth development of functional foods of Rana oil.This project used histomorphological methods to observe the morphological changes of the oviduct in pre-hibernation and the breeding period of the Chinese brown frog(Rana dybowskii).The no-parameter transcriptome experiment was used to detect the enrichment of differential genes in oviduct of the Chinese brown frog in pre-hibernation.Enzyme tissue method was used to detect the content of sex steroid hormone-cholesterol in the oviduct of the Chinese brown frog(Rana dybowskii)in the two periods.The content of progesterone in the oviduct of the Chinese brown frog(Rana dybowskii)in the two periods was determined by enzyme-linked immunosorbent assay.The expression levels of the rate-limiting enzymes ofIVcholesterol synthesis(HMGCR),low-density lipoprotein receptor(LDLR),cholesterol transporter protein(St AR),progesterone synthesis rate-limiting enzyme(P450scc),and transcription factor(SF-1)in oviduct of the Chinese brown frog in two periods was detected through immunohistochemistry,western blot and Real Time PCR experiments.The main results of this study are as follows:(1)The results of histological experiments showed that the glandular cells of the oviduct of the Chinese brown frog(Rana dybowskii)expanded significantly in pre-hibernation compared to breeding period,but the morphology of the epithelial cells did not change significantly.(2)At the level of gene expression,non-parametric transcriptome results showed that the translation and steroid hormone synthesis pathway were significantly overexpressed in pre-hibernation.Real Time PCR experiments showed that the expression levels of HMGCR and LDLR in the oviduct in pre-hibernation were significantly higher than those in the breeding period,while the expression levels of St AR,P450 scc and SF-1 did not show significant changes.(3)At the level of metabolite expression,cholesterol measurement results showed that the content of free cholesterol in oviduct of the Chinese brown frog in pre-hibernation was 11.25±1.8 μmol/g,which was significantly lower than the content of 28.78±2.5 μmol/g in the oviduct in breeding period.The concentration of progesterone in oviduct of the Chinese brown frog in pre-hibernation was 0.782±0.044 ng/m L,which was significantly lower than the content of 1.32±0.21 ng/m L in the oviduct in breeding period.(4)At the level of protein expression,the results of immunohistochemical experiments showed that HMGCR,LDLR,St AR,P450 scc,and SF-1 were expressed in the oviduct in two periods.The results of western blotting experiments showed that the expression levels of HMGCR,LDLR,St AR,and P450 scc in the oviduct of Chinese brown frog in the pre-hibernation were significantly higher than the breeding period,while the expression levels of SF-1 did not show no significant difference.The results of this study showed that the oviduct of Chinese brown frog-Ranaoil had the ability to quickly use cholesterol to synthesize progesterone in the pre-hibernation.The results of this study were significant to the health of specific groups such as menopause and menopause women,and provide a scientific basis for the sustainable development and utilization of Rana oil functional food.