柴黄胃溃宁对胃溃疡肝郁脾虚证模型大鼠血清SOD及HIF-1α、LDH-A影响的实验研究

发布时间：2018-01-19 20:44

本文关键词： 胃溃疡肝郁脾虚证柴黄胃溃宁 SOD HIF-1α LDH-A　出处：《浙江中医药大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的探讨柴黄胃溃宁对胃溃疡肝郁脾虚证模型大鼠胃黏膜低氧微环境是否具有改善作用,通过观察其对SOD及HIF-1a、LDH-A的影响,探讨其可能的作用机制,明确胃溃疡肝郁脾虚证的证本质,进一步推动验方柴黄胃溃宁的临床开发应用。方法将60只雄性SD大鼠在适应性饲养7天后随机分为2组,分别为正常组20只,胃溃疡肝郁脾虚证模型组(简称模型组)40只。采用多种复合因素结合乙酸烧灼法制备大鼠模型,造模结束后分别从正常组和模型组中随机各抽取10只大鼠,以验证胃溃疡肝郁脾虚证模型。验证模型后,将剩余30只模型组大鼠再次随机分为胃溃疡肝郁脾虚证模型组(后亦称模型组)、黄芪建中汤药物干预组(后简称黄芪组)、柴黄胃溃宁药物干预组(后分别简称为柴黄组),每组各10只。正常组剩余10只大鼠继续作为对照组。其中黄芪组与柴黄组于造模结束第2天开始灌服实验方中药水煎剂,连续灌胃14天。各组于末次给药后24h处死,取腹主动脉血采集样本。用酶联免疫法(ELISA)检测血清超氧化物歧化酶(SOD)及缺氧诱导因子-1a(HIF-1a)、乳酸脱氢酶A(LDH-A)的含量。结果1、与正常组大鼠变化比,肝郁脾虚证胃溃疡大鼠模型组体重变化有非常显著性差异(P0.01);其血清GAS、AMY及血浆5-HT较正常组不同程度减低(P0.05或P0.01),血浆NE水平较正常组明显升高(P0.01);其胃窦区黏膜表面颜色较苍白,伴充血水肿,可见被覆薄层渗出物的溃疡面,镜下胃黏膜层有缺损,固有腺体排列较紊乱,主细胞、壁细胞、黏液细胞形态不规则,可见大量炎症细胞浸润,肌层增厚。模型组符合胃溃疡肝郁脾虚证的各项检测指标,表明模型建立成功。2、模型组大鼠HIF-1a、LDH高表达,而SOD则出现低表达。利用柴黄胃溃宁汤剂干预治疗14天后,实验组血清HIF-1a、LDH含量较前降低,SOD活性则有所升高。血清SOD、LDH与HIF-1a与模型组相比,具有显著性统计学差异(P0.05)。结论1、通过模型组大鼠HIF-1a、LDH表达上调,而SOD则出现低表达,进一步明确胃溃疡肝郁脾虚证大鼠模型存在低氧微环境的改变。2、通过中药干预后的HIF-1 a与LDH表达变化,明确柴黄胃溃宁能够调节三羟酸循环能量代谢,改善胃溃疡肝郁脾虚证的低氧微环境;同时表明柴黄胃溃宁可提升机体SOD的表达含量,改善氧自由基代谢调节及低氧应答能力,减轻了脂质过氧化物对机体组织细胞的氧化损伤。3、验方柴黄胃溃宁对胃溃疡肝郁脾虚证大鼠具有良好的改善低氧缺血状态及修复胃黏膜的作用,其机制可能与调节线粒体能量代谢及干预氧自由基代谢有关。
[Abstract]:Objective to investigate whether Chaihuang Weikui Ning can improve gastric mucosal hypoxic microenvironment in rats with gastric ulcer model of liver stagnation and spleen deficiency, and observe its effect on SOD and HIF-1a LDH-A. To explore the possible mechanism of its action, to clarify the nature of the syndrome of stomach ulcer, liver stagnation and spleen deficiency. Methods 60 male Sprague-Dawley rats were randomly divided into 2 groups after 7 days of adaptive feeding. There were 40 rats in the model group of gastric ulcer with liver stagnation and spleen deficiency syndrome (referred to as the model group). The rat model was made by combination of multiple factors and acetic acid cauterization. At the end of the model, 10 rats were randomly selected from the normal group and the model group to verify the model of gastric ulcer, liver stagnation and spleen deficiency. The remaining 30 rats in the model group were randomly divided into the model group of gastric ulcer liver stagnation and spleen deficiency syndrome (later called model group, Huangqi Jianzhong decoction intervention group). Chaihuang Weikuining drug intervention group (later referred to as Chaihuang group). There were 10 rats in each group. The remaining 10 rats in the normal group continued to serve as the control group. The astragalus group and the chaihuang group began to take the experimental prescription Chinese medicine decoction on the second day after the end of the model making. Each group was killed 24 hours after the last administration. Blood samples were collected from abdominal aorta. Serum superoxide dismutase (SOD) and hypoxia inducible factor-1a( HIF-1a) were detected by enzyme linked immunosorbent assay (Elisa). Results 1. Compared with the normal group, the changes of body weight in the model group of gastric ulcer with liver stagnation and spleen deficiency syndrome were significantly different from those in the normal group (P 0.01). The serum level of GASAMY and 5-HT were significantly lower than that of the normal group (P0.05 or P0.01), and the plasma NE level was significantly higher than that of the normal group (P 0.01). Its antral area mucous membrane surface color is paler, accompanied by congestion and edema, can be seen coated with thin layer exudate ulcer surface, under the microscope gastric mucosal layer defects, proper gland arrangement is more disorder, main cells, wall cells. The mucus cells were irregular, a large number of inflammatory cells infiltrated, muscle layer thickened. The model group accord with the various indicators of gastric ulcer liver stagnation and spleen deficiency syndrome, indicating that the model was established successfully. 2. The model group rats HIF-1a. After 14 days of intervention with Chaihuang Weichui Ning decoction, the serum HIF-1a LDH content of the experimental group was lower than that of the former. Compared with the model group, the activity of SOD in serum was significantly higher than that in the model group (P 0.05). Conclusion 1, HIF-1a in the model group was passed through the model group. The expression of LDH was up-regulated, while the expression of SOD was low. It was further confirmed that the rat model of gastric ulcer with liver stagnation and spleen deficiency had the change of hypoxic microenvironment. 2. Through the changes of HIF-1 a and LDH expression after the intervention of traditional Chinese medicine, it is clear that Chaihuang Weikui Ning can regulate the energy metabolism of trihydroxylic acid circulation and improve the hypoxic microenvironment of gastric ulcer with liver stagnation and spleen deficiency syndrome. At the same time, Chaihuang Weikui Ning can improve the expression of SOD, improve the metabolism of oxygen free radicals and the ability of hypoxia response, and alleviate the oxidative damage of lipid peroxide to tissues and cells. 3. Chaihuang Weikui Ning has a good effect on improving hypoxic ischemia and repairing gastric mucosa in rats with gastric ulcer, liver stagnation and spleen deficiency syndrome. The mechanism may be related to regulating mitochondrial energy metabolism and interfering with oxygen free radical metabolism.
【学位授予单位】：浙江中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5;R-332

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