| Objective:To evaluate the efficacy and safety of graphene-based warm uterus acupoint patch in the treatment of primary dysmenorrhoea（PD）through clinical research,and to clarify its clinical efficacy.Through animal experiments,the mechanism of its intervention effect was explored by regulating the hormone and receptor levels of the’hypothalamus-pituitary-ovarian axis’,which laid a theoretical foundation for the popularization and application of graphene-based warm uterus acupoint patch.Methods:1.Clinical initial screening of PD patients for basic information such as age,past medical history,menstrual status,etc.was done through a case screening form.This was followed by a free pre-enrolment physical examination,specifically including routine blood tests,routine urine tests,full liver and kidney tests,electrocardiogram,gynaecological ultrasound,and screening for pregnancy.Next,a randomised double-blind controlled study method was used,in which 74 study subjects who met the inclusion criteria were divided into a control group and a treatment group,each with 37 cases,according to the random numbers given by the randomisation system.Each patient was given three menstrual cycles of intervention treatment and follow-up evaluation by virtue of their own random number.Finally,the COX dysmenorrhea symptom scale score（CMSS）was used as the main outcome index to observe the improvement of dysmenorrhea symptoms,and the pain visual analogue scale（VAS）and the quality of life scale（SF-36）were used as the secondary outcome indicators.The clinical efficacy and safety of graphene-based warm uterus acupoint patch in the treatment of PD were evaluated from the aspects of pain,sleep,life and adverse reactions.At the same time,the blood of the two groups of subjects was collected to detect the expression levels of prostaglandin F2α（PGF2α）,prostaglandin E2（PGE2）,estrogen（E2）and progesterone（PROG）in serum,and the neuroendocrine mechanism of graphene-based warm uterus acupoint patch in the treatment of PD was verified by its expression changes.2.Thirty female SD rats were randomly divided into 3 groups,including control group,model group and treatment group,with 10 rats in each group.The PD rat model was established by using estradiol benzoate combined with oxytocin.After intervention,the writhing latency time,writhing times and writhing scores of each group were observed,and the organ index of uterine tissue was observed.HE staining was used to observe the morphological changes of uterine tissue.Enzyme-linked immunosorbent assay was used to detect the expression of related inflammatory factors（TNF-α,β-EP）,pain factors（PGF2α,OT）and sex hormones（E2,PROG）were detected by ELISA.Western blot and Real-time PCR were used to detect the protein expression and m RNA transcription levels of Gn RH in the hypothalamus,Gn RH-R in the pituitary,and ovarian（FSHR,LHR）in the’hypothalamus-pituitary-ovarian axis’.The protein expression and m RNA transcription levels of progesterone receptor PR and cyclooxygenase-2（COX-2）in uterine tissue were used to explore the mechanism of the intervention effect of graphene warm uterus acupoint sticking on the’hypothalamus-pituitary-ovarian axis’of PD rats.Results:1.Before treatment,the general data of the two groups of patients were evaluated.The results showed that there was no statistical difference in the general data of patients at baseline（P>0.05）,and the data were comparable.2.After treatment,the scores of COX dysmenorrhea symptom scale,pain visual analogue scale and quality of life scale were improved,the content of serum prostaglandin F2αdecreased,and the content of prostaglandin E2 increased.The comparison between the two groups was P<0.05,indicating that both groups were effective for primary dysmenorrhea.The comparison between the two groups showed that the CMSS scores of the treatment group after 3 months of treatment and 3 months of follow-up were statistically different from those of the control group（P<0.05）,indicating that the effect of graphene warm uterus acupoint paste on PD was better than that of placebo paste.3.Safety evaluation:There were no obvious adverse reactions and side effects in the two groups of drugs used in this study,which could be used for clinical treatment of PD patients.4.Torsion response:Compared with the control group,the twisting latency time of the model group was shortened,and the twisting number and twisting scores within 30 min were significantly increased（P<0.01）.Compared with the model group,the twisting latency time of the rats in the treatment group was prolonged,and the twisting number and twisting scores within 30 min were significantly reduced（P<0.01）.5.The organ index of uterine tissue:Compared with the control group,the organ index of uterine tissue in the model group was significantly increased（P<0.01）.Compared with the model group,the organ index of uterine tissue in the treatment group was significantly decreased（P<0.01）.6.Morphological changes of uterine tissue:The structure of endometrial epithelial cells in the control group was clear,neatly arranged,and single-layer columnar.No congestion,edema and obvious inflammatory cell infiltration were found in the glandular cavity and endometrial stroma.In the model group,there were a large number of vacuolar degeneration and apoptosis of endometrial epithelial cells,which were pseudostratified columnar.Gland cavity congestion,endometrial interstitial congestion with edema,and a large number of neutrophil infiltration.It shows that the PD rat model is successful.The endometrial epithelium of rats in the treatment group showed a small amount of vacuolar degeneration and apoptosis,which was high columnar.The degree of congestion and edema in the glandular cavity and endometrial stroma was mild,with only a small amount of neutrophil infiltration.7.ELISA test results:Compared with the control group,the serum levels of TNF-α,PGF2α,OT and E2 in the model group were significantly increased（P<0.01）,and the contents ofβ-EP and PROG were significantly decreased（P<0.01）.Compared with the model group,the serum levels of TNF-α,PGF2α,OT and E2 in the treatment group were significantly decreased（P<0.01）,and the levels ofβ-EP and PROG were significantly increased（P<0.01）.8.Results of Western Blot:Compared with the control group,the protein expression levels of Gn RH,Gn RH-R,FSHR,LHR and PR in the model group were significantly decreased（P<0.01）,and the expression level of COX-2 protein was significantly increased（P<0.01）.Compared with the model group,the protein expression levels of Gn RH,Gn RH-R,FSHR,LHR and PR in the treatment group were significantly increased（P<0.05,P<0.01）,and the protein expression level of COX-2 was significantly decreased（P<0.01）.9.Results of Real-time PCR:Compared with the control group,the relative expression of Gn RH,Gn RH-R,FSHR,LHR and PR m RNA in the model group was significantly decreased（P<0.01）,and the relative expression of COX-2 m RNA was significantly increased（P<0.01）.Compared with the model group,the relative expression of Gn RH,Gn RH-R,FSHR,LHR and PR m RNA in the treatment group was significantly increased（P<0.05,P<0.01）,and the relative expression of COX-2 m RNA was significantly decreased（P<0.01）.Conclusion:1.Graphene-based warm uterus acupoint patch can reduce the severity of pain in PD patients,shorten the duration of pain,regulate hormone levels,and improve the accompanying dysmenorrhea symptoms.The clinical treatment effect is significant.The curative effect is lasting,and there is no obvious adverse reaction,which has clinical research and promotion value.2.Graphene-based warm uterus acupoint patch can effectively inhibit the torsion response of PD rats,reduce the inflammatory edema of uterine tissue,improve the pathological damage of uterine tissue,regulate the contents of inflammatory factors,pain factors and sex hormones,and may play a therapeutic role in PD rats by regulating the levels of hormones and receptors on the uterus and hypothalamus-pituitary-ovarian axis. |
PDF Full Text Request