【摘要】 目的探讨糖尿病胃轻瘫(DGP)发生过程中胃平滑肌组织LC3蛋白与氧化应激-AMPK-TSC2-mTOR通路的变化及意义。方法制备糖尿病大鼠模型,实验分为正常对照组(NC)、模型4周组(DM4w)及模型6周组(DM6w)。检测各组大鼠胃平滑肌组织MDA、T-SOD、CAT的变化。Western blot检测各组大鼠胃平滑肌LC3、AMPK、phospho-AMPK、phospho-TSC2、phospho-mTOR蛋白的表达。结果 DM4w组、DM6w组胃平滑肌组织MDA逐渐增加,且DM6w组与NC组比较,差异有统计学意义(P<0. 05);T-SOD、CAT逐渐下降,且DM6w组及DM4w组与NC组比较,差异均有统计学意义(P<0. 05或P<0. 01)。LC3表现为LC3Ⅰ和LC3Ⅱ两条带,其中,LC3Ⅰ的相对表达量变化不明显,LC3Ⅱ相对表达量逐渐增高(P<0. 01)。AMPK相对表达变化不明显,phospho-AMPK相对表达随病程延长逐渐增加(P<0. 01)。DM4w组、DM6w组phospho-TSC2相对表达与NC组比较,差异均有统计学意义(P<0. 01)。DM4w组phospho-mTOR相对表达较NC组增加(P<0. 01),DM6w组相对表达较DM4w组降低(P<0. 01)。结论 DGP发生过程中,AMPK-TSC2-mTOR参与了氧化应激诱导的胃平滑肌组织自噬的发生,但初期其作用不明显。更多还原

【Abstract】 Objective To explore the changes and value of LC3 protein and oxidative stress--AMPK--TSC2--mTOR pathway in gastric smooth muscle tissue in the development of diabetes gastroparesis.Methods Diabetic model was generated by using Sprague-Dawley(SD)rats. All the SD rats were divided into normal control(NC)group,model four weeks(DM4 w)group and model six weeks(DM6 w)group. The changes of MDA,T-SOD and CAT in gastric smooth muscle were tested by biochemical detection method. The expression of LC3,AMPK,phospho-AMPK,phospho-TSC2 and phospho-mTOR protein were measured by Western blot.Results The MDA in gastric smooth muscle tissue increased gradually in DM4 w and DM6 w groups. And the MDA level was significantly different in DM6 w group compared with NC group(P<0. 05). T-SOD and CAT decreased gradually. And T-SOD and CAT were significantly different in DM6 w and DM4 w group compared with NC group(P<0. 05 or P<0. 01). LC3 presented two bands,LC3 Ⅰ and LC3 Ⅱ. Among them,the relative expression of LC3 Ⅰ did not change significantly,while the relative expression of LC3 Ⅱ increased gradually(P<0. 01). The relative expression of AMPK did not change significantly,and the relative expression of phospho-AMPK gradually increased as the duration of disease increased(P<0. 01). The phospho-TSC2 relative expression was significantly different in DM6 w and DM4 w group compared with NC group(P<0. 01). The relative expression of phospho-m TOR was higher in DM4 w group than in NC group(P<0. 01),and the relative expression was decreased in DM6 w group than in DM4 w group(P<0. 01).Conclusion AMPK-TSC2-mTOR was involved in the oxidative stress-induced gastric smooth muscle autophagy,but its effect was not obvious at the initial stage in the progress of diabetes gastroparesis.更多还原