摘要
目的探讨三七白及散调节磷酸肌醇3激酶(PI3K)/蛋白激酶B(AKT)信号通路对血瘀型胃溃疡(Gastric ulcer,GU)大鼠胃黏膜损伤的影响。方法将50只SPF级健康SD大鼠按随机数字表法分为对照组、模型组、三七白及散低剂量(1.16 g/kg)组、三七白及散高剂量(2.32 g/kg)组、三七白及散高剂量(2.32 g/kg)+740Y-P(PI3K激活剂,10 mg/kg)组,每组各10只,模型组与给药组大鼠采用冷暴露+灌胃盐酸乙醇溶液法构建血瘀型GU模型,以三七白及散与740Y-P分组干预治疗后检测大鼠胃溃疡面积、溃疡指数(Ulcer index,UI)与胃黏膜血流(Gastric mucosal blood flow,GMBF);检测大鼠血液流变学指标全血高切黏度、全血中切黏度、全血低切黏度与血浆黏度;HE染色检测大鼠胃黏膜组织病理形态;ELISA测定大鼠血清促炎因子白细胞介素-6(Interleukin-6,IL-6)、前列腺E2(Prostaglandin E2,PGE2)、肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)水平;用试剂盒检测大鼠胃黏膜组织氧化应激因子谷胱甘肽过氧化物酶(Glutathione peroxidase,GSH-Px)、超氧化物歧化酶(Superoxide dismutase,SOD)、丙二醛(Malondialdehyde,MDA)水平;免疫印迹法检测大鼠胃黏膜组织PI3K/AKT通路相关蛋白表达。结果与对照组比较,模型组大鼠胃黏膜组织有明显病理损伤,胃溃疡面积与UI、全血高切黏度、全血中切黏度、全血低切黏度与血浆黏度、血清IL-6、PGE2与TNF-α水平、胃黏膜组织MDA水平与p-PI3K/PI3K、p-AKT/AKT明显升高(P<0.05),GMBF、胃黏膜组织GSH-Px、SOD水平明显降低(P<0.05)。与模型组比较,三七白及散低、高剂量组大鼠胃黏膜组织病理损伤均减轻,胃溃疡面积与UI、全血高切黏度、全血中切黏度、全血低切黏度与血浆黏度、血清IL-6、PGE2与TNF-α水平、胃黏膜组织MDA水平与p-PI3K/PI3K、p-AKT/AKT均降低(P<0.05),GMBF、胃黏膜组织GSH-Px、SOD水平均升高(P<0.05)。与三七白及散低剂量组比较,三七白及散高剂量组大鼠胃溃疡面积与UI、全血高切黏度、全血中切黏度、全血低切黏度与血浆黏度、血清IL-6、PGE2与TNF-α水平、胃黏膜组织MDA水平与p-PI3K/PI3K、p-AKT/AKT进一步降低(P<0.05),GMBF、胃黏膜组织GSH-Px、SOD水平进一步升高(P<0.05)。与三七白及散高剂量组比较,三七白及散高剂量+740Y-P组大鼠胃溃疡面积与UI、全血高切黏度、全血中切黏度、全血低切黏度与血浆黏度、血清IL-6、PGE2与TNF-α水平、胃黏膜组织MDA水平与p-PI3K/PI3K、p-AKT/AKT升高(P<0.05),GMBF、胃黏膜组织GSH-Px、SOD水平降低(P<0.05)。结论三七白及散可通过抑制PI3K/AKT信号通路激活而减轻血瘀型GU大鼠炎症与氧化应激,改善其血液流变学指标与胃黏膜血液循环,进而减轻大鼠胃黏膜损伤。
Objective To explore the effect of Sanqi Baiji Powder on gastric mucosal injury in rats with blood stasis-type gastric ulcers(GU)by regulating the phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)signaling pathway.Methods Fifty SPF-grade healthy SD rats were randomly divided into control group,model group,low-dose Sanqi Baiji Powder group(1.16 g/kg),high-dose Sanqi Baiji Powder group(2.32 g/kg),and high-dose Sanqi Baiji Powder+740Y-P(PI3K activator,10 mg/kg)group according to a random number table,with 10 rats in each group.The blood stasistype GU model was established using cold exposure and gastric administration of hydrochloric ethanol solution in rats from the model group and the drug treatment groups.Rats were treated with corresponding drugs(Sanqi Baiji Powder and 740Y-P).After treatment,GU area,ulcer index(UI),and gastric mucosal blood flow(GMBF)were measured.Blood rheology parameters including whole blood high,medium,and low shear viscosities,and plasma viscosity were assessed.HE staining was used for histopathological examination of gastric mucosa,and ELISA determined serum pro-inflammatory cytokines,i.e.,interleukin-6(IL-6),prostaglandin E2(PGE2),and tumor necrosis factor-α(TNF-α).Oxidative stress markers in gastric mucosa were measured,including glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),and malondialdehyde(MDA).Western blot detected the expression of PI3K/AKT pathway-related proteins in gastric mucosal tissues.Results Compared with the control group,the model group showed significant pathological damage to gastric mucosa,increased ulcer area,UI,GMBF,whole blood high,medium,and low shear viscosities,,plasma viscosity,serum IL-6,PGE2,TNF-αlevels,MDA levels in gastric mucosal tissues,and p-PI3K/PI3K and p-AKT/AKT ratios(P<0.05),and significantly reduced GSH-Px and SOD levels(P<0.05).Compared with the model group,the low-and high-dose Sanqi Baiji Powder groups showed alleviated pathological injury of gastric mucosa,decreased ulcer area,UI,GMBF,whole blood high,medium,and low shear viscosities,plasma viscosity,serum IL-6,PGE2,TNF-αlevels,MDA levels in gastric mucosal tissues,and p-PI3K/PI3K and p-AKT/AKT ratios(P<0.05),and significantly increased GSH-Px and SOD levels in gastric mucosal tissues(P<0.05).Compared with the low-dose Sanqi Baiji Powder group,the high-dose Sanqi Baiji Powder group showed lower ulcer area,UI,GMBF,whole blood high,medium,and low shear viscosities,plasma viscosity,serum IL-6,PGE2,TNF-αlevels,MDA levels in gastric mucosal tissues,and p-PI3K/PI3K and p-AKT/AKT ratios(P<0.05),and higher GSH-Px and SOD levels in gastric mucosal tissues(P<0.05).Compared with the high-dose Sanqi Baiji Powder group,the high-dose Sanqi Baiji Powder+740Y-P group showed higher ulcer area,UI,GMBF,whole blood high,medium,and low shear viscosities,plasma viscosity,serum IL-6,PGE2,TNF-αlevels,MDA levels in gastric mucosal tissues,and p-PI3K/PI3K and p-AKT/AKT ratios(P<0.05),and lower GSH-Px and SOD levels in gastric mucosal tissues(P<0.05).Conclusion Sanqi Baiji Powder alleviates inflammation and oxidative stress in blood stasis-type GU rats by inhibiting the activation of the PI3K/AKT signaling pathway and improves blood rheological parameters and gastric mucosal circulation,thereby reducing gastric mucosal injury.
作者
王红妹
刘建
李小飞
吴元祥
张飞
WANG Hong-mei;LIU Jian;LI Xiao-fei;WU Yuan-xiang;ZHANG Fei(Department of Spleen and Stomach Diseases,Rugao Hospital of Traditional Chinese Medicine,R ugao Jiangsu 226500)
出处
《世界中西医结合杂志》
2024年第9期1766-1772,共7页
World Journal of Integrated Traditional and Western Medicine
基金
2023年度江苏省中医药科技发展计划项目(MS2023118)
2023年度南通市自然科学基金和社会民生科技计划(指导性)项目(MSZ2023165)。
