| Purpose : In this study,a DOR rat model similar to human was established by intraperitoneal injection of cyclophosphamide.Normal rats were set as the blank group,and the western medicine group,Chinese medicine group,acupuncture group and acupuncture-medicine combination group were respectively set up to treat the DOR rat model.The purpose of this study was to observe the changes of endometrial histological changes before and after treatment in each group of rats,and to detect the level of serum AMH and GLUT4 ?AKT?miR-223 in the endometrium.To explore the effect of acupuncture and drug on the reproductive ability of DOR rats,and to explore the expression of miR-223/ Akt /GLUT4 signaling pathway as a new target for treatment of DOR.Material and method:Sixty healthy female SD rats,aged 8-9 weeks and weighing about 200 g,were selected and fed adaptive for one week.They were divided into six groups by random number table method,including blank group,model group,western medicine group,traditional Chinese medicine group,acupuncture group and acupuncture and medicine group,with 10 rats in each group.Except for blank group,rats in the other five groups were given intraperitoneal injection of cyclophosphamide(50mg/kg/d)for one week after the estrous cycle for modeling.The estrous cycle of rats was observed by vaginal smear method.The stopping or disappearance of a certain phase was considered as successful modeling.The rats were given DHEA intragastric administration,plus or minus Erxian decoction intragastric administration,plus or minus Erxian decoction intragastric administration,plus or minus Erxian decoction intragastric administration,plus or minus Erxian decoction intragastric administration,plus or minus Erxian decoction intragastric administration,plus or minus Erxian decoction intragastric administration,plus or minus Erxian decoction in combination with acupuncture for three consecutive weeks.The morphological changes of endometrium were observed,the serum AMH value,the expression of AKT,GLUT4 and miR-223 in endometrium of rats were detected.Results:1.Hematoxylin-eosin staining results showed that under light microscope,endometrial glands in the blank group were abundant in number,with expanded glandular lumen,abundant folds,large volume,and abundant glandular matrix,arranged in clusters.The number of endometrial glands in the model group was less,and the glandular lumen was small and straight,almost without folds,and the volume was reduced.The glandular structure was abnormal,and there was no cluster arrangement,and the pathological changes were more obvious than those in the normal group.In the western medicine group,the endometrial glands were significantly increased compared with the model group,with enlarged glandular lumen,more folds and increased volume.The structure of the glands was improved compared with the model group,and they were arranged in clusters.In the traditional Chinese medicine group,compared with the model group,the number of endometrial glands was slightly increased,the glandular cavity was enlarged,the folds were increased,the volume was slightly increased,and some of them were arranged in clusters.In the acupuncture group,compared with the model group,the number of endometrial glands was slightly increased,the glandular cavity was slightly expanded,the folds were increased,the volume was slightly increased,and some of them were arranged in clusters.In the acupuncture and medicine group,the endometrial glands were significantly increased compared with the model group.The glandular cavity was enlarged,the folds were increased,and the volume was increased.The glands were arranged in clusters,and the structure was improved significantly compared with the model group.2.The results of enzyme-linked immunosorbent assay showed that compared with blank group,the serum AMH content in model group was significantly decreased(P < 0.05);Compared with model group,the contents of western medicine group,Chinese medicine group,acupuncture group and acupuncture and medicine group were significantly increased(P <0.05);Compared with the traditional Chinese medicine group,the content of acupuncture and medicine group had no statistical significance.Compared with acupuncture group,the content of acupuncture and medicine group was significantly increased(P < 0.05).3.Western blot detection results showed that compared with blank group,the Akt protein content in model group was significantly decreased(P < 0.01);Compared with model group,the contents of western medicine group,Chinese medicine group,acupuncture group and acupuncture and medicine group were significantly increased(P < 0.01);Compared with the traditional Chinese medicine group,the content of the acupuncture-medicine group was significantly increased(P < 0.01),and that of the acupuncture-medicine group was significantly increased(P < 0.01).Compared with blank group,GLUT4 protein content in model group was significantly decreased(P < 0.01);Compared with model group,the white content of western medicine group,Chinese medicine group,acupuncture group and acupuncture and medicine group was significantly increased(P < 0.01);Compared with the traditional Chinese medicine group,the content of acupuncture and medicine group was significantly increased(P < 0.05);Compared with the acupuncture group,the content of acupuncture and medicine group was significantly increased(P < 0.01).4.Real-time quantitative PCR results showed that compared with blank group,the expression level of miR-223 in endometrium of rats in model group was significantly increased(P <0.01);Compared with model group,the expression levels of western medicine group,Chinese medicine group,acupuncture group and acupuncture and medicine group were significantly decreased(P < 0.01);Compared with the traditional Chinese medicine group,the expression level of acupuncture and medicine group was not statistically significant.Compared with acupuncture group,the expression level of acupuncture and medicine group was significantly decreased(P < 0.01).Compared with blank group,the expression level of Akt in model group was significantly decreased(P < 0.01);Compared with model group,the expression levels of western medicine group,Chinese medicine group,acupuncture group and acupuncture and medicine group were significantly increased(P < 0.01);Compared with the traditional Chinese medicine group,the expression level of rats in the acupuncture and medicine group was significantly increased,and that in the acupuncture and medicine group was significantly increased(P < 0.01).Compared with blank group,the expression level of GLUT4 in endometrium of model group was significantly decreased(P < 0.01).Compared with model group,the expression levels of western medicine group,Chinese medicine group,acupuncture group and acupuncture and medicine group were significantly increased(P < 0.01);Compared with the traditional Chinese medicine group,the expression level of rats in the acupuncture and medicine group was significantly increased(P < 0.01),and that in the acupuncture and medicine group was significantly increased(P < 0.01).Conclusion:1.The combination of acupuncture and medicine can improve the endometrial histological changes of DOR and regulate the level of serum AMH.2.The effect of acupuncture and medicine combined with DOR in rats was better than that of traditional Chinese medicine or acupuncture alone.3.Acupuncture and drug combination can improve endometrial receptivity and ovarian reserve function of DOR rats by regulating miR-223/ Akt /GLUT4 signaling pathway. |
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