基于氧化应激介导的SCF/c-Kit系统探讨养阴和胃方治疗糖尿病胃轻瘫的机制研究

Diabetic gastroparesis (DGP) is one of common chronic complications of diabetes, Which is caused by gastrointestinal hormone disorders, gastric smooth muscle degeneration, and hyperglycemia, etc. In recent years, as described, most of the known and central mechanisms of Diabetic gastroparesis (DGP) is one of common chronic complications of diabetes. Recent studies show that oxidative stress and SCF/c-kit system have critical roles in the pathogenesis of DGP. Yangyin Hewei formula has shown a significant effect in the treatment of DGP, but the exact mechanism remains unclear. Our previous study shows that this formula can effectively improve gastric emptying and gastric blood flow, meanwhile, it can increase the expression of interstitial cells of Cajal (ICC) in gastric antrum of DGP rats. Whether Yangyin Hewei formula protects ICCs in DGP rats by increasing SCF/c-Kit expression based on oxidative stress and normalizing the delay in gastric emptying needs to be verified by further experiments. The project intends to establish STZ induced DGP rats, in order to evaluate the efficacy of Yangyin Hewei formula depending on the analysis of gastric emptying rates. Simultaneously, Observing the expression of interstitial cells of Cajal(ICC)、c-kit、SCF in the gastric tissue and the expression of MDA、SOD、8-OH-dG、HO-1 in the blood and gastric tissue, We clarify the treatment mechanism of Yangyin Hewei formula on DGP. It may provide a new idea on seeking new drugs for DGP.

糖尿病胃轻瘫（DGP）是糖尿病常见而难治的慢性并发症之一，近来，氧化应激及SCF/c-kit系统在DGP发病中的地位逐渐引起重视。养阴和胃方是根据胃阴不足、脾气虚弱的病机而定，临床上治疗DGP疗效显著，但其具体机制尚不明确。课题组前期研究发现，该方能增强DGP大鼠的胃动力和胃血流量，同时增加DGP大鼠胃窦Cajal间质细胞的含量。但养阴和胃法是否通过氧化应激介导的SCF/c-kit系统来改善Cajal间质细胞以及网络结构，从而改善胃排空，仍有待实验进一步验证。本项目拟建立DGP大鼠模型，通过酚红灌胃实验和离体肌条测定评价养阴和胃法治疗DGP的疗效，同时，通过检测胃组织Cajal间质细胞、c-Kit、SCF的表达，以及血液和胃组织内MDA、SOD、8-OH-dG、HO-1的表达，进一步阐明养阴和胃法治疗DGP的作用机理，为临床提供新的治疗思路和方法。

糖尿病胃轻瘫（DGP）是糖尿病常见而难治的慢性并发症之一，不仅会导致患者诸多症状，且消耗大量的医疗保健资源，严重影响患者的生活质量。DGP目前缺乏有效治疗，养阴和胃方是根据胃阴不足、脾气虚弱的病机而定，临床上治疗DGP疗效显著，但其具体机制尚不明确。本课题选择STZ干预的DGP大鼠模型作为研究对象，将养阴和胃方分为低、高剂量组进行给药，同时选择枸橼酸莫沙必利混悬液作为对照药物，观察养阴和胃方对DGP大鼠的胃功能的影响， 通过流式细胞术检测Cajal间质细胞凋亡，Western blotting检测自噬相关蛋白Caspase-3和Caspase-9的表达，免疫印迹法检测细胞增殖标志蛋白Ki67的表达法，用IHC法检测各组福尔马林固定石蜡包埋胃组织中c-kit的表达，通过western blotting和qRT-PCR法分别检测各组胃组织中c-kit的蛋白和mRNA表达。采用酶联免疫吸附法测定各组大鼠血清SCF蛋白浓度。通过化学比色法测定胃组织SOD活性、MDA含量，通过western blotting和qRT-PCR法分别检测各组大鼠胃组织内蛋白和mRNA表达。研究结果显示，养阴和胃方组大鼠的胃排空、胃运动、胃蠕动和胃血流量均优于DGP模型组，养阴和胃方组胃组织Cajal细胞凋亡率明显下降， caspase-3、caspase-9和Ki67的表达明显下降，而c-kit的蛋白、mRNA表达，血清SCF蛋白表达均明显改善，养阴和胃方组大鼠SOD表达较DGP模型组显著升高，MOD表达较DGP模型组显著降低， HO-1的蛋白和mRNA表达均明显改善。以上结果养阴和胃方可通过提示养阴和胃方可通过改善高氧化应激状态，改善HO-1水平，上调SCF/c-kit系统的表达，降低Cajal间质细胞的凋亡，从而逆转DGP胃排空障碍。