文献类型：期刊文献

中文题名：基于MSK1/2探讨少腹逐瘀汤治疗子宫内膜异位症痛经寒凝血瘀证的作用机制

英文题名：Mechanism of Shaofu Zhuyu Decoction in treatment of endometriosis-associated dysmenorrhea with syndrome of cold coagulation and blood stasis based on MSK1/2

作者：陈元欢[1];毛海燕[1];武权生[1];张小花[1];申剑[1];冯鹏[1];黄灿灿[1];吉秀家[1]

第一作者：陈元欢

机构：[1]甘肃中医药大学中医临床学院,甘肃兰州730000

第一机构：甘肃中医药大学中医临床学院

年份：2022

卷号：47

期号：17

起止页码：4674

中文期刊名：中国中药杂志

外文期刊名：China Journal of Chinese Materia Medica

收录：CSTPCD;;Scopus;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；PubMed;

基金：国家自然科学基金项目(81760879);甘肃中医药大学科学研究与创新基金项目(30140419);2018年甘肃省中医药管理局科研项目(GZK-2018-1)。

语种：中文

中文关键词：子宫内膜异位症;寒凝血瘀证;痛经;少腹逐瘀汤;MSK1;MSK2;DUSP1;IL-6;IL-10;PGE2;TNF-α

外文关键词：endometriosis;syndrome of cold coagulation and blood stasis;dysmenorrhea;Shaofu Zhuyu Decoction;MSK1;MSK2;DUSP1;IL-6;IL-10;PGE2;TNF-α

摘要：基于丝裂原和应激激活蛋白激酶1/2(mitogen-and stress-activated protein kinase 1/2,MSK1/2)的作用机制探讨少腹逐瘀汤对子宫内膜异位症(endometriosis,EMT)痛经寒凝血瘀证大鼠的干预作用及其机制研究。借助随机数字表从SPF级雌性未孕大鼠中随机选出假手术组,其余大鼠采用“自体移植法+冰箱冷冻法”进行EMT寒凝血瘀证造模,造模成功后随机分为对照组及少腹逐瘀汤高、中、低剂量组。少腹逐瘀汤高、中、低剂量组分别以9、4.5、2.3 g·kg^(-1)灌胃,每日1次,连续2周;对照组药物选取孕三烯酮0.24 mg·kg^(-1)灌胃,每3 d 1次,连续2周。2周后,开腹测量各组大鼠异位灶大小;酶联免疫吸附测定(enzyme-linked immuno sorbent assay,ELISA)法测量血清中白细胞介素(interleukin,IL)-6、IL-10、前列腺素E2(prostaglandin E2,PGE2)、肿瘤坏死因子-α(tumor necrosis factorα,TNF-α)表达,蛋白免疫印迹(Western blot)法检测大鼠在位子宫内膜组织中MSK1/2、双特异性磷酸酶-1(dual-specificityphosphatases,DUSP1)蛋白表达,使用实时荧光定量聚合酶链式反应(real-time PCR,RT-PCR)法检测大鼠在位子宫内膜组织中MSK1/2、DUSP1 mRNA表达。经ELISA检测发现,与假手术组相比,模型组大鼠血清中IL-6、PGE2、TNF-α水平明显升高(P<0.01),IL-10水平明显降低(P<0.01);与模型组相比,少腹逐瘀汤高、中剂量组及孕三烯酮组大鼠血清中IL-6、PGE2、TNF-α水平明显降低(P<0.01),IL-10水平明显升高(P<0.01)。经Western blot检测得出,与假手术组相比,模型组大鼠在位子宫内膜组织中MSK1、MSK2、DUSP1蛋白表达明显降低(P<0.01);与模型组相比,少腹逐瘀汤高、中剂量组及孕三烯酮组大鼠在位子宫内膜组织中MSK1、MSK2、DUSP1蛋白表达明显升高(P<0.01)。通过RT-PCR得到,与假手术组相比,模型组大鼠在位子宫内膜组织中MSK1、MSK2、DUSP1 mRNA表达明显降低(P<0.01);与模型组相比,少腹逐瘀汤高剂量组及孕三烯酮组大鼠在位子宫内膜组织中MSK1、MSK2、DUSP1 mRNA表达明显升高(P<0.01)。少腹逐瘀汤可通过影响MSK1、MSK2蛋白,调节促炎因子TNF-α、IL-6、PGE2及抗炎因子IL-10、DUSP1的异常表达,改善EMT痛经寒凝血瘀证大鼠体内的炎症反应,从而对EMT痛经寒凝血瘀证大鼠起到治疗作用。
This study aims to decipher the mechanism underlying the effect of Shaofu Zhuyu Decoction on endometriosis(EMT)-associated dysmenorrhea in rats with the syndrome of cold coagulation and blood stasis based on mitogen-and stress-activated protein kinase 1/2(MSK1/2).We employed a random number table to randomly assign SPF female non-pregnant rats into the sham group,and treated the rest rats with autologous transplantation+refrigerator freezing for the modeling of the syndrome of cold coagulation and blood stasis.The modeled rats were then randomly assigned into the control group and high-,medium-and low-dose Shaofu Zhuyu Decoction groups.The rats in the low-,medium-,and high-dose decoction groups were respectively administrated with 9,4.5,and 2.3 g·kg^(-1)decoction through gavage once a day for 2 consecutive weeks,and those in the control group were administrated with 0.24 mg·kg^(-1)gestrinone through gavage once every 3 days for 2 weeks.After that,the size of ectopic focus in each rat was measured via laparotomy.Enzyme-linked immunosorbent assay(ELISA)was adopted to determine the expression of interleukin(IL)-6,IL-10,prostaglandin E2(PGE2),tumor necrosis factor-α(TNF-α).Western blot was employed to determine the protein levels of MSK1/2 and dual-specificity phosphatase 1(DUSP1)and real-time quantitative polymerase chain reaction(RT-PCR)to determine the mRNA levels of the two genes in rat eutopic endometrial tissue.Compared with the sham group,the model group showed increased levels of IL-6,PGE2,and TNF-αwhile decrease level of IL-10 in the serum(P<0.01).Compared with the model group,the high-and medium-dose decoction groups and the gestrinone group had declined levels of IL-6,PGE2,and TNF-αwhile risen level of IL-10 in the serum(P<0.01).The model group had lower protein levels and mRNA levels of MSK1/2 and DUSP1 in the eutopic endometrial tissue than the sham group(P<0.01).The high-and medium-dose decoction groups and the gestrinone group had higher protein and mRNA levels of MSK1/2 and DUSP1 in the eutopic endometrial tissue than the model group(P<0.01).The results indicated that Shaofu Zhuyu Decoction can regulate the abnormal expression of pro-inflammatory cytokines TNF-α,IL-6,and PGE2 and anti-inflammatory cytokines IL-10 and DUSP1 via MSK1/2 to alleviate EMT-associated dysmenorrhea in rats with the syndrome of cold coagulation and blood stasis.