葛根芩连汤及配伍调控Nrf2/NQO1信号通路抑制溃疡性结肠炎大鼠氧化应激损伤  

Gegen Qinliantang and Its Combinations Inhibit Oxidative Stress Injury in Ulcerative Colitis Rats by Regulating Nrf2/NQO1 Signaling Pathway

作　　者：林川 王菲 王鸿卿 王新宏[1] 安叡[1] LIN Chuan;WANG Fei;WANG Hongqing;WANG Xinhong;AN Rui(School of Pharmacy,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学中药学院,上海201203

出　　处：《中国实验方剂学杂志》2022年第13期19-27,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：上海市“科技创新行动计划”自然科学基金项目(19ZR1451900)。

摘　　要：目的:探讨葛根芩连汤治疗溃疡性结肠炎(UC)大鼠的可能机制,同时讨论缺少药味对葛根芩连汤疗效的影响。方法:采用自由饮用5%葡聚糖硫酸钠盐水溶液诱导UC大鼠模型,雄性SD大鼠随机分为正常组、模型组、西药组(柳氮磺吡啶肠溶片,350 mg·kg^(-1))、葛根芩连汤全方组(17 g·kg^(-1))、全方去甘草组(17 g·kg^(-1))、全方去葛根组(17 g·kg^(-1))、葛根甘草组(17 g·kg^(-1))和黄芩黄连组(17 g·kg^(-1))。用1,1-二苯基-2-苦肼基(DPPH)、2,2-联氨-双(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS)和荧光漂白后恢复技术(FRAP)法评估葛根芩连汤及其不同配伍组体外抗氧化活性;通过比较大鼠体质量变化和记录结肠长度、观察结肠剖面和苏木素-伊红(HE)染色病理组织切片评估给药治疗后各组大鼠结肠组织损伤程度;比色法测定大鼠血清中髓过氧化物酶(MPO)、脂质过氧化物(LPO)、丙二醛(MDA)、总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)和还原型谷胱甘肽(GSH)水平变化;实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)分别检测结肠组织中核因子E_(2)相关因子2(Nrf2)、醌氧化还原酶1(NQO1)和血红素氧合酶-1(HO-1)mRNA和蛋白表达的变化。结果:与正常组比较,模型组大鼠结肠黏膜组织坏死,炎症浸润,血清中过氧化物MPO、LPO和MDA含量显著上升(P<0.01),抗氧化酶T-SOD、CAT和GSH活性显著下降(P<0.01),Nrf2、NQO1和HO-1 mRNA表达均呈下降趋势,Nrf2蛋白表达显著下降(P<0.01),NQO1和HO-1蛋白表达呈下降趋势;与模型组比较,葛根芩连汤及配伍组能改善UC大鼠结肠组织病理损伤和形态结构,显著降低大鼠血清中过氧化物MPO、LPO和MDA水平(P<0.05),提高无葛根组抗氧化酶T-SOD,无甘草组、黄芩黄连组CAT和GSH(P<0.01)活性,增加结肠组织中Nrf2、NQO1和HO-1 mRNA和蛋白的相对表达,全方组差异明显(P<0.05)。结论:葛根芩连汤对UC大鼠病理损伤Objective:To explore the underlying mechanism of Gegen Qinliantang(GGQL)in the treatment of ulcerative colitis(UC)in rats and discuss the effects of modification of GGQL on its efficacy.Method:The UC model was induced in rats by free access to 5%dextran sulfate sodium in saline solution.Male SD rats were randomly divided into a normal group,a model group,a positive control group(sulfasalazine enteric-coated tablets,350 mg·kg^(-1)),a GGQL group(17 g·kg^(-1)),a Glycyrrhizae Radix et Rhizoma(GR)-absent GGQL group(17 g·kg^(-1)),a Puerariae Lobatae Radix(PLR)-absent GGQL group(17 g·kg^(-1)),a GRPLR group(17 g·kg^(-1)),and a Scutellariae Radix(SR)-Coptidis Rhizoma(CR)group(17 g·kg^(-1)).The in vitro antioxidant activities of GGQL and its combinations were evaluated by 2,2-diphenyl-1-picrylhydrazyl(DPPH),2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS),and fluorescence recovery after photobleaching(FRAP)methods.The degree of colonic tissue injury in each group was evaluated based on the weight changes of rats,the length of the colon,the colon sections,and hematoxylin-eosin(HE)-stained histopathologic sections.The serum levels of myeloperoxidase(MPO),lipid peroxide(LPO),malondialdehyde(MDA),total superoxide dismutase(T-SOD),catalase(CAT),and reduced glutathione(GSH)were measured by colorimetry.The mRNA and protein expression of nuclear factor-erythroid 2 related factor(Nrf2),quinone oxidoreductase 1(NQO1),and heme oxygenase-1(HO-1)in colon tissues was detected by real-time fluorescence-based quantitative polymerase chain reaction(Real-time PCR)and Western blot,respectively.Result:Compared with the normal group,the model group showed colonic mucosal necrosis,inflammatory infiltration,increased serum levels of MPO,LPO,and MDA(P<0.01),blunted activities of T-SOD,CAT,and GSH(P<0.01),decreasing trend of mRNA expression of Nrf2,NQO1,and HO-1,reduced expression of Nrf2 protein(P<0.01),and decreasing trend of expression of NQO1 and HO-1 proteins.Compared with the model group,the GGQL and its combination group

关 键 词：葛根芩连汤 不同配伍 氧化应激 溃疡性结肠炎 核因子E2相关因子2(Nrf2)信号通路 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R284R33