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目的:探讨脾虚型功能性消化不良(FD)大鼠胃组织线粒体呼吸链复合物IV亚单位(COX VA)蛋白和mRNA表达及脾虚一号方对其的干预作用。方法:70只7 d龄雄性SD大鼠随机分为正常组,FD模型组(单模型组),脾虚型FD模型组(双模型组),多潘立酮组,脾虚一号方低、中、高剂量组,每组10只。FD模型组、脾虚证FD模型组给予0.1%蔗糖碘乙酰胺蔗糖溶液灌胃,连续6 d。脾虚证FD模型组正常饲料喂养至6周龄后叠加改良小平台站立,连续14 d。造模结束后分别给予蒸馏水10 m L·kg~(-1)·d~(-1),多潘立酮3.125 mg·kg~(-1)·d~(-1),脾虚一号方1.275,2.55,5.1 g·kg~(-1)·d~(-1),灌胃14 d。采用实时荧光定量聚合酶链式反应(Real-time PCR),蛋白质免疫印迹(Western blot)和免疫组化法检测胃窦组织COX VA mRNA与蛋白表达量。结果:与正常组比较,脾虚一号方高剂量组COX VA蛋白平均积分吸光度升高(P<0.05);多潘立酮组、脾虚一号方低、中剂量组COX VA蛋白表达量均有不同程度降低(P<0.01),而脾虚一号方高剂量组COX VA蛋白表达量则升高(P<0.01);单模型组COX VA mRNA表达量升高最明显(P<0.01)。与双模型组比较,多潘立酮组、脾虚一号方低、中剂量组COX VA蛋白表达量均有不同程度降低(P<0.01),多潘立酮组、脾虚一号方低、中剂量组COX VA mRNA表达量均有不同程度降低,而脾虚一号方高剂量组、单模型组则升高,差异均无统计学意义。结论:脾虚型FD大鼠存在COX VA蛋白的表达量降低,脾虚一号方能够增加COX VA蛋白的表达量,改善能量代谢。
Objective: To investigate the protein and mRNA expression of mitochondrial respiratory chain complex IV subunit (COX VA) in the stomach of patients with functional dyspepsia (FD) on spleen asthenia and the effect of spleen deficiency one on it. Methods: Seventy male SD rats (7 days old) were randomly divided into normal group, FD model group (single model group), FD model group (double model group), domperidone group, Group, 10 in each group. FD model group and FD model group of spleen deficiency syndrome were given intragastrically with 0.1% sucrose iodoacetamide sucrose solution for 6 days. Spleen Deficiency FD model group normal diet until 6 weeks of age after the superposition of a small platform to stand for 14 days. After the model was finished, the rats were given 10 m L · kg -1 d -1 of distilled water, 3.125 mg · kg -1 d -1 of droperidone, 1.275,2.55 , 5.1 g · kg -1 (-1) d -1 for 14 days. Real-time PCR, Western blot and immunohistochemistry were used to detect the mRNA and protein expression of COX VA in gastric antrum. Results: Compared with the normal group, the mean integral absorbance of COX VA increased (P <0.05) in the high-dose Spleen-No.1 Prescription group; the COX VA protein expression decreased in the domperidone group and spleen deficiency-1 low dose group (P <0.01), while the expression of COX VA protein in spleen deficiency high-dose group increased (P <0.01). The expression of COX VA mRNA increased significantly in single model group (P <0.01). Compared with the model group, the expression of COX VA protein in the domperidone group and the low and medium dose group of Spleen deficiency No.1 decreased to some extent (P <0.01) The levels of spleen deficiency were significantly higher than those of high-dose spleen-deficiency group and single-model group, but the difference was not statistically significant. Conclusion: The expression of COX VA protein in spleen deficiency type FD rats is decreased. Spleen No.1 Formula can increase the expression of COX VA protein and improve the energy metabolism.