扶正消萎汤对慢性萎缩性胃炎大鼠胃黏膜修复作用及Nrf2/ARE通路的影响研究

Effect of FuzhengXiaowei Decoction on gastric mucosal repair and Nrf2/ARE pathway in chronic atrophic gastritis rats

目的研究扶正消萎汤对慢性萎缩性胃炎(CAG)大鼠胃黏膜修复作用及核转录因子2(Nrf2)/抗氧化反应元件(ARE)通路的影响。方法将大鼠随机分为空白对照组及造模组,采用1-甲基-3-硝基-1-亚硝基胍(MNNG)联合乙醇灌胃法建立CAG模型,造模完成后,大鼠随机分为CAG组、扶正消萎汤低、中、高剂量组及阳性对照组,每组10只,扶正消萎汤低、中、高剂量组分别按照0.74、1.48、2.22 g/kg(以生药含量计)灌胃给予大鼠扶正消萎汤,阳性对照组按照0.24 g/kg灌胃给予大鼠维酶素,连续干预6周,测定干预前后各组大鼠体质量变化,使用酶联免疫吸附法(ELISA)试剂盒血清环氧合酶(COX)-2、诱导型一氧化氮合酶(i-NOS)、白介素(IL)-6、肿瘤坏死因子(TNF)-α水平及胃黏膜一氧化氮(NO)、一氧化氮合成酶(NOS)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化脂质(LPO)、丙二醛(MDA)、超氧化物歧化酶(SOD)水平,苏木精-伊红(HE)染色法检查大鼠胃黏膜病理学变化,实时定量聚合酶链式反应(RTq-PCR)测定大鼠胃黏膜Nrf2、ARE及HO-1 mRNA水平,免疫印迹法测定胃黏膜Nrf2、ARE及HO-1蛋白水平。结果与空白对照组比较,CAG组大鼠体质量、胃黏膜组织GSH-Px及SOD水平、胃组织Nrf2、ARE及HO-1 mRNA及蛋白水平显著降低(P<0.05),大鼠胃黏膜NO、NOS、LPO及MDA水平、血清COX-2、i-NOS、IL-6及TNF-α水平显著升高(P<0.05);与CAG组比较,扶正消萎汤各剂量组及阳性对照组大鼠体质量、胃黏膜组织GSH-Px及SOD水平、胃组织Nrf2、ARE及HO-1 mRNA及蛋白水平显著升高(P<0.05),大鼠胃黏膜NO、NOS、LPO及MDA水平、血清COX-2、i-NOS、IL-6及TNF-α水平显著降低(P<0.05),大鼠胃组织病理学改变明显恢复,且随着扶正消萎汤剂量的增加,各项指标变化更优。结论扶正消萎汤能够显著抑制慢性萎缩性胃炎大鼠疾病进展,修复胃组织病理性改变,抑制炎症反应及氧化应激损伤,其机制可能与调节Nrf2/ARE通路有关。

Objective To study the effects of Fuzheng Xiaowei Decoction on gastric mucosal repair and nuclear tran-scription factor 2(Nrf2)/antioxidant reaction element(ARE)pathway in chronic atrophic gastritis(CAG)rats.Methods Rats were randomly divided into blank control group and modeling group,and CAG model was established by 1-methyl-3-nitro-1-nitroguanidine(MNNG)combined with ethanol gavage.After modeling,rats were randomly divided into CAG group,FuzhengXiaowei decoction low-dose,medium-dose and high-dose groups and positive control group,with 10 rats/group.The rats in the low,medium and high dose groups of FuzhengXiaowei Decoction were administrated by intragastric administration of 0.74 g/kg,1.48 g/kg and 2.22 g/kg(measured by crude drug content),respectively,while the rats in the positive control group were administrated by intragastric administration of 0.24 g/kg with vitase for 6w.The changes of body mass in each group before and after intervention were measured.The serum levels of cyclooxygenase(COX-2),inducible nitric oxide synthase(i-NOS),interleukin-6,tumor necrosis factor(TNF-α)and gastric mucosa nitric oxide(NO),nitric oxide synthase(NOS),glutathione peroxidase(GSH-Px)were used by enzyme-linked immunosorbent assay(ELISA)The levels of oxidized lipids(LPO),malondialdehyde(MDA),superoxide dismutase(SOD),hematoxylin-eosin(HE)staining were used to detect the pathological changes of gastric mucosa in rats,and the mRNA levels of Nrf2,ARE and HO-1 in gastric mucosa were determined by real-time quantitative polymerase chain reaction(RTq-PCR).The levels of Nrf2,ARE and HO-1 in gastric mucosa were determined by Western blotting.Results Compared with blank control group,body weight,GSH-Px and SOD levels of gastric mucosa,Nrf2,ARE and HO-1 mRNA and protein levels of gastric tissue in CAG group were significantly decreased(P<0.05).The levels of NO,NOS,LPO and MDA in gastric mucosa and the levels of COX-2,i-NOS,IL-6 and TNF-α in serum were significantly increased(P<0.05).Compared with CAG group,body weight,GSH-Px and SOD levels of gastric mucosa,Nrf2,ARE and HO-1 mRNA and protein levels of gastric tissue in FuzhengXiaowei Decoction dose groups and positive control group were significantly increased(P<0.05).The levels of NO,NOS,LPO and MDA in gastric mucosa,COX-2,i-NOS,IL-6 and TNF-αin serum of rats were significantly decreased(P<0.05),and the gastric histomathological changes of rats were significantly recovered,and with the increase of the dose of FuzhengXiaowei Decoction,the changes of various indexes were better.Conclusion FuzhengXiaowei Decoction can significantly inhibit the progression of chronic atrophic gastritis in rats,repair the pathological changes of gastric tissue,inhibit inflammation and oxidative stress damage,and its mechanism may be related to the regulation of Nrf2/ARE pathway.