| Objective: The effectiveness of graphene-based warm uterus acupoint paste in the treatment of primary dysmenorrhea(PD)was observed to explore its transcriptomic mechanism of treating dysmenorrhea by RNA-seq.Methods:1.PD patients were screened and baseline information was collected for age,menstrual cycle,days of menstruation and years of education before enrollment,and then were included and baseline physical examination was performed after meeting the inclusion criteria.A randomized controlled double-blind trial design was used,in which 60 numbers were randomly generated in the background and divided into treatment and control groups,with third-party personnel affixing the corresponding numbers after grouping to self-sealing bags with the same appearance.Once patients were enrolled,they were randomly selected by the experimental personnel according to the 60 numbers that had been generated and were applied to CV 4,EX-CA1(double)and SP6(double).It is applied seven days before menstruation for a total of ten days,one month as a course of treatment,for a total of three courses of treatment.The Dysmenorrhea Symptom Rating Scale was evaluated for efficacy.The degree of menstrual pain was evaluated by Visual Analogue Score(VAS),menstrual symptoms were evaluated by Cox Menstrual Symptom Scale(CMSS)before treatment and after each session,Beck Anxiety Inventory(BAI),Beck Depression Inventory-II(BDI-II)and Quality of Life Score(Short Form 36 Body Pain Scale,SF-36)were evaluated before treatment and after the last session.2.There were 18 SD female uncrossed rats,aged between 6 and 8 weeks,randomly divided into blank group(6),model group(6)and treatment group(6).The mice were modeled using the classical dysmenorrhea modeling method for a total of 13 days.On the4 th day of modeling,the lower abdomen and the Sanyinjiao point of the mice were applied for 10 days.On day 14,after abdominal anesthesia,biological information(differential genes,GO,KEGG,etc.)of PD rats were screened and enriched for analysis using gene sequencing technology.Results:1.Regarding the overall efficacy,6(20.69%)were significantly effective,14(48.28%)were effective,and 9(31.03%)were ineffective in the treatment group,with an overall effective rate of 68.97%,and 7(25.93%)were effective in the control group,with an overall effective rate of 25.93%.2.Compared with before treatment: VAS and CMSS(duration)scores in the treatment group decreased in the second course(P<0.05)and in the third course(P<0.01),CMSS(pain level)decreased in the third course(P<0.05),BAI and BDI-II scores decreased after the third course(P<0.01),and SF-36 scores increased after the third course(P<0.01),and the changes of each index score in the control group were not statistically significant(P>0.05);compared with post-treatment: the scores of VAS in the treatment group were lower than those in the control group after the second course(P<0.05)and the third course(P<0.001),the scores of CMSS(duration)were lower than those in the control group after the second course(P<0.01)and the third course(P< 0.001),CMSS(pain level)score after the third course was lower than that of the control group(P<0.01),BAI and BDI-II scores after the third course were lower than that of the control group(P<0.01),and all SF-36 scores after the third course were higher than that of the control group(P<0.01)3.Differential gene screening: There were 2537 genes changed in the model vs.blank group,including 1007 up-regulated genes and 1530 down-regulated genes;387 genes changed in the treatment vs.model,including 39 up-regulated genes and 348down-regulated genes,were co-contained with 81 differential genes.4.GO classification enrichment: the most enriched genes in the treatment group BP were involved in the cell cycle and the highest enrichment index was involved in the biological process of mitotic sister chromatid segregation nuclear division;the most enriched genes in MF were with the set of signal receptors and the highest enrichment index was with the nucleus binding function;in CC,the most enriched genes were with the presence of chromosomes and the highest enrichment index was with the The presence of condensed nuclear chromosomes gene nuclei,condensed nuclear chromosomes,and centromeric regions.The most enriched gene in control BP is involved in systemic development,the highest enrichment index is positive regulation of developmental processes,the most enriched gene in MF is signal receptor binding,the highest enrichment index is microtubule binding,the most enriched gene in CC is intrinsic component of membrane the highest enrichment index is extracellular matrix.5.KEGG enrichment: 215 KEGG PATHWAYs were enriched in the treatment group,mainly in Cellular processes,Human diseases,Metabolism,Biological systems,Environmental information processing and related to PD,such as Vascular smooth muscle contraction pathway,Arachidonic acid pathway,c AMP signaling pathway,Calcium signaling pathway,renin secretion,etc.;322 KEGG PATHWAYs were enriched in the model group,divided into five categories,such as cellular processes,were enriched to the same6-regulated pathways as the treatment group.Finally,combined with the results of PPI interactions,it was found that graphene warming patch for PD was closely related to Vascular smooth muscle contraction,Linoleic acid metabolism,Arachidonic acid metabolism and Calcium pathway.Conclusion:1.Graphene-based warm uterus acupoint paste efficiently reduces pain and menstrual symptoms are improved,anxiety and depression are relieved,and the quality of life is improved.2.In the treatment of PD rats,the mechanism of graphene-based warm uterus acupoint paste was achieved by regulating the m RNA expression of pla2g2 a,calml3 down-regulation affecting Vascular smooth muscle contraction pathway,Arachidonic acid metabolic pathway,Linoleic acid metabolism and Calcium pathway. |
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