基于Nrf2/HO-1通路探讨二十五味鬼臼丸对宫颈炎大鼠的治疗作用及机制  

Study on therapeutic effects and mechanism of Ershiwuwei Guijiu pills on cervicitis rats based on Nrf2/HO-1 pathway

作　　者：张赫林 龚春香 赵解南 朱彦蓉 谢和兵 袁志兵 ZHANG Helin;GONG Chunxiang;ZHAO Jienan;ZHU Yanrong;XIE Hebing;YUAN Zhibing(School of Pharmacy,Anhui University of Chinese Medicine,Hefei 230013,China;Jiangsu Shenhou Pharmaceutical Research Co.,Ltd.,Nantong 226133,China;Department of Pathology,Changzhou Traditional Chinese Medicine Hospital affiliated to Nanjing University of Chinese Medicine,Changzhou 213003,China;Rexplo Medical Laboratory of Guangxi Medical University,Nanning 530000,China)

机构地区：[1]安徽中医药大学药学院,安徽合肥230013 [2]江苏神猴医药研究有限公司,江苏南通226133 [3]南京中医药大学附属常州中医医院病理科,江苏常州213003 [4]广西医科大学睿谷医学检验实验室,广西南宁530000

出　　处：《药物评价研究》2023年第12期2608-2616,共9页Drug Evaluation Research

基　　金：国家自然科学基金资助项目(81470247)。

摘　　要：目的研究藏药二十五味鬼臼丸对苯酚胶浆致宫颈炎模型大鼠的治疗作用及机制。方法将雌性SD大鼠48只随机分为对照组、模型组及二十五味鬼臼丸低、中、高剂量(0.2、0.4、0.8 g·kg^(-1))组和抗宫炎片(0.47 g·kg^(-1))组,每组8只。除对照组外,其余5组建立25%苯酚胶浆致宫颈炎大鼠模型,各药物组分别以相应药物ig给药,每日1次,连续给药12 d后取材。观察大鼠外阴炎症程度评分、子宫指数、子宫解剖形态学改变;采用苏木精-伊红(HE)染色观察大鼠子宫组织病理学改变;采用酶联免疫吸附测定(ELISA)法检测子宫组织中超氧化物歧化酶(SOD)、丙二醛(MDA)含量变化;采用实时荧光定量PCR(q RT-PCR)法和蛋白免疫印迹(Western blotting)法检测子宫组织中核因子E2相关因子(Nrf2)、血红素氧合酶-1(HO-1)、NADPH氧化酶4(Nox4)的m RNA和蛋白表达情况。结果与对照组比较,模型组大鼠外阴炎症评分、子宫指数、子宫组织中MDA水平、Nox4的m RNA和蛋白表达均显著升高(P<0.01),体质量、SOD活性、Nrf2、HO-1的m RNA和蛋白表达均显著降低(P<0.01),子宫组织出现不同程度的病理变化;与模型组比较,二十五味鬼臼丸能明显降低外阴炎症评分、子宫指数、子宫组织中MDA水平、Nox4的m RNA和蛋白表达(P<0.05、0.01),升高体质量及子宫组织中SOD活性和Nrf2、HO-1的m RNA和蛋白表达(P<0.05、0.01),减轻子宫颈炎症损伤程度。结论二十五味鬼臼丸可能通过抑制Nox4的表达,激活Nrf2/HO-1通路,发挥抗氧化作用,减轻炎症反应,从而抑制宫颈炎发展。Objective To study the therapeutic effects and mechanism of Tibetan medicine Ershiwuwei Guijiu pills(EGP)on cervicitis model rats induced by phenol mucilage.Methods Totally 48 female SD rats were randomly divided into control group,model group,EGP low,medium and high dose group(0.2,0.4,0.8 g·kg^(−1))and Kanggongyan Tablets group(0.47 g·kg^(−1)),with eight rats in each group.Except for control group,the rats in the other five groups were successfully established with 25%phenol mucilage cervicitis,and then the rats in each drug group were given the corresponding drugs by gavage once a day for 12 consecutive days.The vulvar inflammation degree score,uterine index,and uterine anatomical morphological changes were observed.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes of the uterus in rats.The contents of superoxide dismutase(SOD)and malondialdehyde(MDA)in uterine tissues were detected by enzyme-linked immunosorbent(ELISA).The expression of mRNA and protein in uterine tissues by nuclear factor erythroid 2-related factor(Nrf2),heme oxygenase-1(HO-1),and NADPH oxidase 4(Nox4)were detected by real-time fluorescent quantitative PCR(qRT-PCR)and Western blotting.Results Compared with control group,the vulvar inflammation score,uterine index,the content of MDA,the expression of Nox4 mRNA and protein in uterine tissue of the model group were significantly increased(P<0.01),the activity of SOD and the expression of Nrf2,HO-1 mRNA and protein were significantly decreased(P<0.01),and different degrees of pathological changes were observed in uterine tissue;Compared with model group,each dose of EGP group could significantly decrease the vulva inflammation score,uterine index,the content of MDA,the expression of Nox4 mRNA and protein in uterine tissue(P<0.05 and 0.01),and increase the activity of SOD and the expression of Nrf2,HO-1 mRNA and protein in uterine tissue(P<0.05 and 0.01).Conclusion EGP may inhibit the development of cervicitis by inhibiting the expression of Nox4,activating the

关 键 词：二十五味鬼臼丸 宫颈炎 苯酚胶浆 核因子E2相关因子 氧化应激 

分 类 号：R285.5[医药卫生—中药学]