摘要
目的研究半夏泻心汤对溃疡性结肠炎(UC)小鼠氧化应激损伤的影响,并探讨核转录因子E2相关因子(Nrf2)/血红素加氧酶-1(HO-1)通路在其中发挥的作用。方法2023年1―5月,小鼠自由饮用3%葡聚糖硫酸钠(DSS)1周,建立溃疡性结肠炎模型。60只雄性C57BL/6J小鼠采用随机数字表法分为空白组、模型组、美沙拉嗪组(100 mg/kg)、半夏泻心汤高、中、低剂量组(32.76、16.38、8.19 g/kg)。造模同时灌胃给予相应药物,连续10 d。实验结束时称量小鼠体质量、测量结肠长度并观察结肠组织病理学改变。酶联免疫法检测血清中活性氧自由基(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-px)变化情况;RT-PCR和蛋白质印迹法分别检测Nrf2、HO-1、醌氧化还原酶-1(NQO-1)的mRNA和蛋白含量。结果空白组小鼠结肠长度、ROS、MDA、SOD、GSH-px含量分别为(12.53±0.94)cm、(9.39±0.76)U/mL、(4.32±0.37)nmol/L、(124.79±11.27)U/mL、(1102.92±87.20)U/mL;模型组小鼠结肠长度、ROS、MDA、SOD、GSH-px含量分别为(5.63±0.45)cm、(13.30±1.47)U/mL、(7.65±0.81)nmol/L、(80.31±8.98)U/mL、(823.00±89.63)U/mL。与空白组相比,模型组小鼠体质量和结肠长度明显降低,结肠黏膜组织隐窝结构被破坏、杯状细胞减少、炎性细胞浸润明显,血清中ROS、MDA含量明显增加,抗氧化酶SOD、GSH-px活性明显降低,Nrf2、HO-1、NQO-1 mRNA及蛋白含量显著下降。半夏泻心汤高剂量组小鼠结肠长度、ROS、MDA、SOD、GSH-px含量分别为(8.97±1.20)cm、(10.49±1.04)U/mL、5.72±0.57 nmol/L、112.24±10.85 U/mL、1032.20±117.66 U/mL;半夏泻心汤中剂量组小鼠结肠长度、ROS、MDA、SOD、GSH-px含量分别为(8.03±0.69)cm、(11.23±1.11)U/mL、(6.47±0.30)nmol/L、(93.20±11.47)U/mL、(993.96±96.72)U/mL;半夏泻心汤低剂量组小鼠结肠长度、ROS、MDA、SOD、GSH-px含量分别为(7.53±0.69)cm、(12.33±1.45)U/mL、(7.23±0.65)nmol/L、(84.89±9.07)U/mL、(891.06±86.61)U/mL。与模型组相比,半夏泻心汤干预后,小鼠体质量和结肠长度明显增加,结肠组织隐窝结构清晰且炎性细胞浸润减少,血清中ROS和MDA含量明显降低,SOD和GSH-px活性明显增加,结肠组织中Nrf2、HO-1、NQO-1 mRNA和蛋白表达亦显著提高。结论半夏泻心汤可改善UC症状,通过调节Nrf2/HO-1相关蛋白表达水平,进而影响氧化应激指标ROS、MDA、SOD和GSH-px含量,减轻结肠病理损伤程度。为半夏泻心汤开发为防治UC药物奠定一定基础。
Objective To study the effect of Banxia Xiexin decoction on oxidative stress injury in ulcerative colitis mice and explore the role of nuclear factor erythroid 2 related factor(Nrf2)/heme oxygenase-1(HO-1)pathway.Methods From January to May 2023,Mice were given 3%dextran sulfate sodium(DSS)for a week to establish the ulcerative colitis model.Sixty male C57BL/6J mice were randomly divided into control group,model group,mesalazine group(100 mg/kg),Banxia Xiexin decoction high-dose,medium-dose and low-dose groups(32.76,16.38 and 8.19 g/kg).At the same time,corresponding drugs were given by gavage for 10 consecutive days.At the end of the experiment,the body weight and colon length of mice were measured,and the pathological change of colon was observed.Serum reactive oxide species(ROS),malondialdehyde(MDA),superoxide dismutase(SOD)and glutathione peroxidase(GSH-px)were detected by enzyme-linked immunosorbent assay.The mRNA and protein expressions of Nrf2,HO-1 and NADPH quinine oxidoreduc-tase-1(NQO-1)were detected by RT-PCR and Western blotting.Results The colon length,ROS,MDA,SOD,GSH-px content of mice in the control group was(12.53±0.94)cm,(9.39±0.76)U/mL,(4.32±0.37)nmol/L,(124.79±11.27)U/mL,and(1102.92±87.20)U/mL,re-spectively.The colon length,ROS,MDA,SOD,GSH-px content of mice in the model group was(5.63±0.45)cm,(13.30±1.47)U/mL,(7.65±0.81)nmol/L,(80.31±8.98)U/mL,and(823.00±89.63)U/mL,respectively.Compared with the control group,the body weight and colon length were significantly reduced in the model group,the crypt structure of colonic mucosal tissue was destroyed,goblet cells were reduced,inflammatory cell infiltration was obvious,the contents of ROS and MDA in serum were significantly increased,and the activi-ties of antioxidant enzymes SOD and GSH-px were significantly decreased.The mRNA and protein expressions of Nrf2,HO-1 and NQO-1 were obviously reduced.The colon length,ROS,MDA,SOD,GSH-px content of mice in the high-dose of Banxia Xiexin decoction group was(8.97±1.20)cm,(10.49±1.04)U/mL,(5.72±0.57)nmol/L,(112.24±10.85)U/mL,and(1032.20±117.66)U/mL,respectively.The colon length,ROS,MDA,SOD,GSH-px content of mice in the medium-dose of Banxia Xiexin decoction group was(8.03±0.69)cm,(11.23±1.11)U/mL,(6.47±0.30)nmol/L,(93.20±11.47)U/mL,and(993.96±96.72)U/mL,respectively.The colon length,ROS,MDA,SOD,GSH-px content of mice in the low-dose of Banxia Xiexin decoction group was(7.53±0.69)cm,(12.33±1.45)U/mL,(7.23±0.65)nmol/L,(84.89±9.07)U/mL,and(891.06±86.61)U/mL,respectively.Compared with the model group,after the intervention of Banxia Xiexin decoction,the body weight and colon length of mice were significantly increased,the crypt structure of colon tissue was clear,the infiltration of inflammatory cell was reduced,the contents of ROS and MDA in serum were significantly decreased,and the activities of SOD and GSH-px were re-markably increased.The mRNA and protein expressions of Nrf2,HO-1 and NQO-1 in colon tissues were also significantly increased.Conclusions Banxia Xiexin decoction could improve UC symptoms,regulate the expression level of Nrf2/HO-1 related proteins,then affect the contents of oxidative stress indicators,such as ROS,MDA,SOD and GSH-px,and finally reduce the degree of pathological dam-age of colon.This paper laid a foundation for the development of Banxia Xiexin decoction for the prevention and treatment of UC.
作者
汪敏
黄亚威
黄然
WANG Min;HUANG Yawei;HUANG Ran(Department of Pharmacy,Jiangsu Province Hospital of Chinese Medicine,Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing,Jiangsu 210029,China)
出处
《安徽医药》
CAS
2024年第5期894-899,I0003,共7页
Anhui Medical and Pharmaceutical Journal
基金
江苏省研究生科研实践计划项目(SJCX21-0717)。
关键词
半夏泻心汤
溃疡性结肠炎
氧化应激
核转录因子E2相关因子
Banxia Xiexin decoction
Ulcerative colitis
Oxidative stress
Nuclear factor erythroid 2 Related factor
