摘要
目的:选取氧化偶氮甲烷/葡聚糖硫酸钠(AOM/DSS)诱导的慢性肠炎癌变小鼠模型,动态探究具有清热燥湿、调气和血功效的芍药汤调控抗氧化应激信号通路Keap1/Nrf2/ARE发挥抗氧化作用对肠"炎-癌"转化进展的保护机制。方法:病理学评价AOM/DSS诱导的肠炎癌变情况;定量RT-PCR检测结直肠组织中抗氧化信号通路核因子E-2-相关因子2(Nrf2)关键指标的表达;免疫组化检测结直肠组织炎症和肿瘤形成指标核因子κB(NF-κB)、Ki-67抗原(Ki-67)、Nrf2的表达;ELISA法检测结直肠组织炎性细胞因子肿瘤坏死因子-α(TNF-α)和白细胞介素1β(IL-1β)以及氧化应激蛋白超氧化物歧化酶(SOD)和丙二醛(MDA)的表达。结果:与AOM模型组比较,芍药汤各组小鼠体质量减轻较少、均能明显抑制结肠缩短、显著减少肿瘤数量(P<0.01)。与AOM组比较,芍药汤高剂量组各时间点谷胱甘肽还原酶(GR)、硫氧还蛋白还原酶(TR)、血红素氧合酶-1(HO-1)、苷二磷酸葡萄糖醛酸基转移酶1A1(UGT1A1)的mRNA表达水平,第63、98天时,醌NADH脱氢酶1(NQO-1)mRNA及第28、63天时尿苷二磷酸葡萄糖醛酸基转移酶1A10(UGT1A10)、γ-谷氨酰半胱氨酸合成酶c(γ-GCSc)mRNA表达水平均显著升高(P<0.05,P<0.01);芍药汤各剂量组第98天NF-κB、Ki-67表达水平显著降低(P<0.01),第63、98天TNF-α及IL-1β表达水平显著降低(P<0.05,P<0.01);芍药汤高、中剂量组各时间点SOD活性增强(P<0.05,P<0.01),芍药汤各剂量组各时间点MDA含量减少(P<0.01)。结论:芍药汤对AOM/DSS诱导的肿瘤发生具有一定程度的预防作用,能够抑制氧化应激诱导的炎性损伤,通过调控抗氧化应激信号通路Keap1/Nrf2/ARE发挥抗氧化和抗炎作用。
Objective:To select a mouse model of chronic enteritis induced by AOM/DSS,and to dynamically explore the Shaoyao Decoction(which has the effect of regulating-qi and dampness-eliminating)protection mechanism of intestinal‘inflammatory-cancer’transformation progression exerted by regulating antioxidative stress signaling pathway Keap1/Nrf2/ARE to produce antioxidation effect on intestinal inflammation.Methods:Pathological evaluation of AOM/DSS-induced colitis transformation to carcinoma;Quantitative RT-PCR for detection of key markers of Nrf2 in antioxidant signaling pathway in colorectal tissues;Immunohistochemical detection for expression of NF-κB,Ki-67 and Nrf2 inflammation and tumor formation related indicators in colorectal tissues;ELISA detection for expression of inflammatory cytokines TNF-α,IL-1βand oxidative stress proteins SOD,MDA in intestinal tissue.Results:Compared with the AOM/DSS model group,the animals in Shaoyao Decoction groups had less weight loss,significantly inhibited colonic shortening and significantly reduced the number of tumors(P<0.01).Compared with the AOM group,the mRNA expression levels of GR,TR,HO-1 and UGT1 A1 in the high dose group of Shaoyao Decoction at each time point,NQO1 mRNA at 63 d and 98 d,and UGT1 A10,γ-GCSc mRNA expression levels at 28 d and 63 d were significantly increased(P<0.05,P<0.01).At 98 d,all of the Shaoyao Decoction groups significantly decreased the expression of NF-κB and Ki-67(P<0.01).At 63 d and 98 d,all of the Shaoyao Decoction groups significantly inhibited the expression of TNF-αand IL-1β(P<0.05,P<0.01).At each time point,the activity of SOD in the high-dose group and the midiumdose group of Shaoyao Decoction was enhanced(P<0.05,P<0.01),and all of the Shaoyao Decoction groups significantly inhibited the increase of MDA content(P<0.01).Conclusion:SYD has a certain degree of preventive effect on AOM/DSS-induced tumorigenesis;SYD can inhibit the oxidative stress-induced inflammatory damage SYD can regulate the antioxidant stress signaling pathway Keap1/Nrf2/ARE to exert antioxidant and anti-inflammatory effects.
作者
王晓燕
李卫东
花宝金
WANG Xiao-yan;LI Wei-dong;HUA Bao-jin(Department of Oncology,Guang'anmen Hospital.China Academy of Chinese Medical Sciences,Beijing 100053,China;Department of Medicare Management,Wangjing Hospital,China Academy of Chinese Medical Sciences,Beijing 100102,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2019年第11期5141-5147,共7页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金面上项目(No.81673906)
中国博士后科学基金第61批面上资助项目(No.2017M611124)~~
