| Clinical partObjective:This study attempts to investigate clinical efficacy and safety of strengthening Yuan Qi and dredging meridians massage on treating decreased ovarian reserve（DOR）based on the HPO axis.Methods:Seventy-two patients with DOR admitted from January 2021 to December2021 were selected into the study.They were randomly divided into control group and experimental group according to the order of visits,with thirty-six patients in each group.The control group took oral estradiol valerate tablets（Progynova,1 mg/d,21d）and diflexion progesterone tablets（Duphaston,20 mg/d,10d）,and the treatment lasted two menstrual cycles.Besides the two medicines above,experimental group was treated with strengthening Yuan Qi and dredging meridians massage simultaneously.The acupoints include Qi Hai,Guan Yuan,Zhong Ji,Gui Lai,Zi Gong and Luan Chao,once a day for six days consecutively,till the end of the western medicine treatment cycles.The menstrual situation,traditional Chinese medicine（TCM）syndrome score and serum basic hormone levels were observed and compared before and after treatment.Bilateral antral follicle count（AFC）,ovarian volume（OV）and the patient’s ovarian interstitial artery flow signal were calculated and measured by vagina ultrasound test,among the 2nd to the 5th days during the menstrual cycle.The efficacy was assessed by the percentage of TCM syndrome and adverse events were monitored during treatment.Follow-up visits lasted for 6 months after treatment and recurrence was observed and recorded.Results:A total of 72 patients with DOR were included in this study,3 withdrew in the experimental group during treatment and follow-up visits,and 33 patients completed the trial.Two withdrew in the control group,and 34 patients completed the trial.Menstrual index scores and TCM symptom scores were significantly lower compared with the scores of pre-treatment（P<0.05）in both groups;compared with control group,the volume,color,quality,menstrual score and TCM symptom scores were significantly lower（P<0.05）in experimental group,and no statistical difference was shown in menstrual cycle and period between the two groups（P>0.05）.In experimental group,FSH,E2levels,FSH/LH ratio and PI and RI indexes,S/D ratio were significantly lower after treatment（P<0.05）,P and PSV,EDV values significantly increased（P<0.05）compared with control group.There was no difference in LH,AMH,AFC and OV（P>0.05）between the two groups,but experimental group shows more obvious progress in recovery after treatment.The treatment efficacy of experimental group（90.9%）was significantly higher than that of control group（70.6%）,and the group comparisons were statistically significant（X2=4.422,P<0.05）.During the 6-month follow-up visits,the recurrence rate in experimental group was much lower,which was15.15%（5/33）,compared with 29.41%（10/34）in control group.Neither group had serious adverse effects during treatment,indicating a high safety overall.Conclusion:The combined treatment of strengthening Yuan Qi and dredging meridians massage and western medicine cycle treatment on DOR can significantly enhance the curative efficacy and reduce the recurrence rate,effectively lessen the severity of menstrual disorder symptoms and ameliorate serum basic hormone levels of DOR patients.In addition,it increases the blood flow perfusion of ovarian interstitial artery,effectively improves pelvic microcirculation,so as to promote the recovery of ovarian reserve function.The method has high safety and clinical application.ExperimentObjective:The efficacy of strengthening Yuan Qi and dredging meridians massage on the reproductive system of rats was observed by DOR rat model.Based on animal experiments,the efficacy and mechanism of strengthening Yuan Qi and dredging meridians massage on the improvement of ovarian reserve function were discussed,providing a scientific basis for the application of strengthening Yuan Qi and dredging meridians massage.Methods:The DOR rat model was established by intragastric administration of triperygium wilfordii multiglucoside suspension,and the changes in the estrous cycle of DOR rats were detected.Rats with successful modeling were randomly divided into 5groups:control group without intervention,model group,strengthening Yuan Qi and dredging meridians massage group,western medicine group,and strengthening Yuan Qi and dredging meridians massage combined with western medicine group.Each group was treated continuously for 21 days according to the norm.The estrous cycle,the morphological structure of the uterus and ovary,the grade and number of follicles,the expression of E2,LH,AMH,FSH,as well as the expression levels of Gn RH,E2,LH,AMH and FSH in the tissue samples were detected after treatment.According to the above indicators,the therapeutic efficacy of strengthening Yuan Qi and dredging meridians massage on DOR rats was evaluated comprehensively.Furthermore,RNA sequencing technology was used to obtain the differential genes of the ovaries in rats between strengthening Yuan Qi and dredging meridians massage group and model group.The GO function enrichment,KEGG pathway enrichment,Reactome analysis and PPI network construction were performed to analyze potential treatment mechanism of strengthening Yuan Qi and dredging meridians massage on rats.Results:Most of the rats in the DOR model group were depressed,unresponsive,and showed estrous cycle disorder.The microscopic structure results showed significant reduction of follicle numbers and follicular dysplasia in the ovary of DOR rats.The uterine structure showed that the distribution of glands was significantly reduced,and the uterine epithelial cells were accumulated,indicating that the rat model of DOR was successfully constructed.Through therapeutic intervention,it was found that strengthening Yuan Qi and dredging meridians massage could improve the structure and function of ovary and uterus in DOR rats,increase the number of growing follicles,reduce the levels of serum LH and FSH（P<0.05）,increase the levels of AHM and E2（P<0.05）,and decrease the expression level of Gn RH in the hypothalamus（P<0.05）.Strengthening Yuan Qi and dredging meridians massage combined with western medicine had the best therapeutic efficacy,and the efficacy of strengthening Yuan Qi and dredging meridians massage was similar to that of western medicine.The experimental results showed that strengthening Yuan Qi and dredging meridians massage can regulate the"hypothalamus-pituitary-ovarian"axis,thereby improving the regulation for the DOR rats.In addition,RNA sequencing and bioinformatics analysis showed that strengthening Yuan Qi and dredging meridians massage can improve DOR through multiple targets and multiple pathways,among which the arachidonic acid metabolic pathway was highly correlate and gene-enriched.RT-q PCR and Western blot analyses showed that strengthening Yuan Qi and dredging meridians massage can significantly increase the m RNA and protein expression of COX-2 and ALOX15 proteins（P<0.05）,and activate the arachidonic acid metabolic pathway;on the contrary,the western medicine treatment did not regulate this metabolic pathway.Conclusion:Animal experiments show that strengthening Yuan Qi and dredging meridians massage alone or combined with western medicine can improve ovarian reserve function,follicle number and oocyte quality in DOR rats.One of the mechanisms by which strengthening Yuan Qi and dredging meridians massage improves DOR is by activating the arachidonic acid metabolic pathway. |
