摘要
目的观察隔物灸对原发性痛经(PD)模型大鼠子宫组织DNA甲基转移酶(DNMT)和脑源性神经营养因子(BDNF)启动子区DNA甲基化的影响,探讨隔物灸对PD模型大鼠表观遗传调控的作用机制。方法将32只雌性SD大鼠随机分成空白组、模型组、隔物灸组和西药组,每组8只。采用冰水浴、苯甲酸雌二醇复合缩宫素方法制备PD寒湿凝滞证大鼠模型,造模第1日起,隔物灸组予隔盐灸“神阙”、隔姜灸“关元”,每穴20 min,西药组予布洛芬溶液灌胃,均每日1次,连续10 d,第11日腹腔注射缩宫素后观察大鼠扭体反应并计算扭体评分,Western blot和荧光定量PCR检测子宫组织BDNF、DNMT3A、DNMT3B蛋白及mRNA表达,免疫组化染色检测子宫组织DNMT3A、DNMT3B阳性表达,亚硫酸盐测序检测子宫组织BDNF启动子区DNA甲基化程度。结果与空白组比较,模型组大鼠扭体潜伏期缩短,扭体评分升高(P<0.01);子宫组织BDNF、DNMT3A、DNMT3B蛋白及mRNA表达升高(P<0.01),DNMT3A、DNMT3B阳性表达升高(P<0.01),BDNF启动子区DNA甲基化率降低(P<0.01)。与模型组比较,隔物灸组和西药组大鼠扭体潜伏期延长,扭体评分降低(P<0.05,P<0.01);子宫组织BDNF、DNMT3A、DNMT3B蛋白及mRNA表达降低(P<0.05,P<0.01),DNMT3A、DNMT3B阳性表达降低(P<0.01),BDNF启动子区DNA甲基化率升高(P<0.01)。结论隔物灸“神阙”“关元”可能通过升高BDNF启动子区DNA甲基化水平抑制BDNF转录,使BDNF表达下调,达到缓解PD模型大鼠疼痛的目的。
Objective To investigate the effects of indirect moxibustion on the expressions of DNA methyltransferases(DNMT)and methylation of the brain-derived neurotrophic factor(BDNF)promoter region in uterine tissues of rats with primary dysmenorrhea(PD);To explore the mechanism of epigenetic regulation of indirect moxibustion on PD model rats.Methods A total of 32 female SD rats were randomly divided into blank group,model group,indirect moxibustion group and Western medicine group,with 8 rats in each group.The PD model with cold dampness stagnation syndrome was established using ice-water baths combined with estradiol benzoate and oxytocin.Starting from the first day of modeling,the indirect moxibustion group received salt-partitioned moxibustion at“Shenque”and ginger-partitioned moxibustion at“Guanyuan”for 20 min,while the Western medicine group was gavaged ibuprofen solution.Both interventions were given once a day for 10 days.On day 11,writhing responses were observed and scored after oxytocin injection,Western blot and RT-qPCR were used to detect protein and mRNA expression of BDNF,DNMT3A and DNMT3B in uterine tissue,immunohistochemical staining was used to detect the positive expressions of DNMT3A and DNMT3B in uterine tissue.The DNA methylation of BDNF promoter region in uterine tissue was detected by sulfite sequencing.Results Compared with the blank group,the writhing latency was shortened and the writhing score increased in the model group(P<0.01);the protein and mRNA expressions of BDNF,DNMT3A and DNMT3B in uterine tissue increased(P<0.01),the positive expressions of DNMT3A and DNMT3B increased(P<0.01),and the DNA methylation rate in BDNF promoter region decreased(P<0.01).Compared with the model group,the writhing latency was lengthened and the writhing score decreased in the indirect moxibustion group and Western medicine group(P<0.05,P<0.01);the protein and mRNA expressions of BDNF,DNMT3A and DNMT3B in uterine tissue decreased(P<0.05,P<0.01),the positive expressions of DNMT3A and DNMT3B decreased(P<0.01),and the DNA methylation rate in BDNF promoter region increased(P<0.01).Conclusion Indirect moxibustion at“Shenque”and“Guanyuan”may inhibit the transcription of BDNF by increasing the DNA methylation level of BDNF promoter region,and reduce the expression of BDNF,so as to relieve the pain of PD rats.
作者
邢彦雪
周敏
王晓妍
毕阔
王迪
赵志国
杨丽芸
张选平
张岩
高飞
李文丽
李新华
XING Yanxue;ZHOU Min;WANG Xiaoyan;BI Kuo;WANG Di;ZHAO Zhiguo;YANG Liyun;ZHANG Xuanping;ZHANG Yan;GAO Fei;LI Wenli;LI Xinhua(School of Acupuncture-Moxibustion and Tuina,Hebei University of Chinese Medicine,Shijiazhuang 050200,China;Hebei International Joint Research Center for Dominant Diseases in Chinese Medicine and Acupuncture,Shijiazhuang 050200,China)
出处
《中国中医药信息杂志》
2025年第9期113-119,共7页
Chinese Journal of Information on Traditional Chinese Medicine
基金
河北省自然科学基金(H2022423368、H2023423004)
河北中医药大学燕赵医学项目(YZZY2022003、YZZY2023018、XCXZZSS2024008)。
