| Objective The research selected to examine by microRNA gene chip screening of differentially expressed gene in PCOS endometrial tissue after oraling tanshinone capsules 3 months. We try to explore the effect of tanshinone capsules, cryptotanshinone and berberine which could improve serum hormone level of PCOS rat model, and regulate the expression of gene and protein signals in endometrium, and the effect of endometrial carcinogenesis related genes with 17 beta estradiol (17β-E2) induced with PCOS rat model.Methods The research selected cases from Huaian Maternal and Child Health Hospital into the group of PCOS patients with insulin resistance, and oral tanshinone capsules 3 months. We curettaged endometrial tissue before and after as the research object, examined by microRNA gene chip screening of differentially expressed gene in PCOS. There were 450 female Sprague-Dawley rats with age of 90 days which induced by Poretsky scheme of subcutaneous injection with insulin and human chorionic gonadotropin.When PCOS rats model were success, treatment groups were gavaged for 4 weeks. Animals were decapitated on the last day of treatment, approximately 12 hours after the last injection. Trunk blood was collected in heparinized polypropylene tubes for measurement of serum hormone levels. Estrogen receptor a protein expression of endometrial tissue was detected using immunohistochemical method. The expression level of AKT3,RAC 1,PGR,Kras and PTEN mRNA gene in endometrium were detected by qRT-PCR.17beta estradiol induced endometrial carcinogenesis in PCOS rat model was Kyoko scheme, collected endometrial tissue and detected by qRT-PCR. The expression level of AKT3, RAC1, PGR, Kras and PTEN mRNA gene in endometrium were detected by qRT-PCR, Independent samples t test, single factor analysis of variance and chi-square test were used for statistical analysis, and P<0.05 was statistically significant.Results (1) There were 7 genes detected differentially. There were 3 down-regulated genes, including hsa-miR-149-5p, hsa-miR-664b-3,hsa-miR-6763-3p; and 4 up-regulated genes, including hsa-miR-630,hsa-miR-1273g-3, hsa-miR-6746-5, hsa-miR-6774-5p.(2)The control group showed average 4-5 days regular estrous cycle, and PCOS model group showed irregular estrous cycle, to prolong the period of more than 5 consecutive days, showing a large number of small round cells, a small amount of keratinized epithelial cells and nucleated epithelial cells and mucus. The treatment groups such as tanshinone capsules group,cryptotanshinone group, berberine group and metformin group indicated that the estrous cycle was gradually restored.(3)There was not obvious difference in body weight before treatment.After treatment with insulin and HCG, there was significant difference in body weight in control group compared with the model group. Compared with model group, tanshinone capsules group, cryptotanshinone group,berberine group and metformin group had obvious difference after treatment,and the body weight decreased significantly in treatment groups.(4)There were many different levels of follicles in control group and the corpus luteum showed by HE staining. There were about 9 layers of granulosa cells in the dominant follicle. In model group, there were a great number of large follicles with cystic changes, as well as atresia follicles, in which the basic structure of the dominant follicle,such as oocytes and corona. The number of granulosa cells were decreased and a large number of corpus luteum in model group. In treatment groups, there was decrease in the number of follicles with cystic changes. Under the HE staining of the uterus, the uterine cavity of the control group was large, with many folds, and the glands were arranged in clusters. In model group, the endometrial glandular cavity was small and straight, with almost no folds, and the number of glands was reduced compared with the control group. In treatment groups,the endometrial glandular cavity was larger than that in model group, and the amount of glands were increased compared with model group.(5) The positive expression of estrogen receptor a protein in endometrial tissue in model group was significantly higher than control group,and the positive expression of estrogen receptor a protein in endometrial tissue in treatment groups were significantly decreased compared with model group.(6) The serum level of testosterone in model group was significantly increased than control group, and the level of testosterone in treatment groups were decreased compared with model group.(7) Compared with control group, the gene expression of AKT3 mRNA was increased in PCOS model group. Compared with PCOS model group, the gene expression of AKT3 mRNA was decreased in tanshinone,cryptotanshinone and berberine groups. Compared with PCOS model group,the gene expression of AKT3 mRNA in metformin group was higher than the gene expression in model group. Compared with control group, the gene expression of RAC1 mRNA was increased in PCOS model group. Compared with PCOS model group, the gene expression of RAC1 mRNA was decreased in treatment groups. Compared with control group, the gene expression of PGR mRNA was decreased in PCOS model group. Compared with PCOS model group, the gene expression of PGR mRNA was increased in treatment groups. Compared with control group, the gene expression of Kras mRNA was increased in PCOS model group. Compared with PCOS model group, the gene expression of Kras mRNA in tanshinone capsules group was higher than the gene expression in model group, but there was no statistical significance.Compared with PCOS model group, the gene expression of Kras mRNA was decreased in cryptotanshinone, berberine and metformin groups. Compared with control group, the gene expression of PTEN mRNA was increased in PCOS model group. Compared with PCOS model group, the gene expression of PTEN mRNA in tanshinone group was higher than the gene expression in model group. Compared with PCOS model group, the gene expression of PTEN mRNA was decreased in cryptotanshinone, berberine and metformin groups. Compared with control group, the gene expression of AKT3 mRNA was increased in 17 beta estradiol induced PCOS rat model group.Compared with 17 beta estradiol induced PCOS rat model group, the gene expression of AKT3 mRNA was decreased in treatment groups. Compared with control group, the gene expression of RAC1 mRNA was increased in 17 beta estradiol induced PCOS rat model group. Compared with 17 beta estradiol induced PCOS rat model group, the gene expression of RAC1 mRNA was decreased in treatment groups. Compared with control group, the gene expression of PGR mRNA was decreased in 17 beta estradiol induced PCOS rat model group. Compared with 17 beta estradiol induced PCOS rat model group, the gene expression of PGR mRNA was increased in tanshinone capsules group, but there was no statistical significance. Compared with 17 beta estradiol induced PCOS rat model group,the gene expression of PGR mRNA was increased in cryptotanshinone, berberine and metformin groups.Compared with control group, and the gene expression of PTEN mRNA was increased in 17 beta estradiol induced PCOS rat model group. Compared with 17 beta estradiol induced PCOS rat model group, the gene expression of PTEN mRNA was decreased in treatment groups.Conclusion The beneficial effect of tanshinone capsules could ameliorate remarkably molecular mechanism by microRNA gene chip screening in PCOS. There were significant effects on the endometrium of PCOS rat model in tanshinone capsules group, cryptotanshinone and berberine groups, which regulate the hormone synthesis and cell proliferation.There was a certain impact to mechanism of tanshinone capsules,cryptotanshinone and berberine on the carcinogenesis related genes in PCOS endometrial tissues. |
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