脾虚一号方对脾虚型功能性消化不良大鼠胃组织线粒体呼吸链复合物Ⅳ亚单位的影响

Effect of Pixu Ⅰ Recipe on Cytochrome C Oxidase Subunit VA Expression in Gastric Tissues of FD Rats with Spleen Deficiency

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 levels of mitoehondrial respiratory chain complex Ⅳ A （COX VA） in gastric tissues of functional dyspepsia （FD） rats with spleen deficiency and explore the effect of Pixu Ⅰ recipe intervention. Method： Seventy 10-day-old male SD rats were randomly divided into normal group, FD model group （single model group） , spleen deficiency type FD model group （dual model group） , Domperidone group, Pixu I recipe low dose group, medium dose group, and high dose group. The rats in FD model group and spleen deficiency type FD model group were garaged with 0.2 mL of 0. 1% IA in 2% sucrose for 6 days. Normal feeding was given to the rats in spleen deficiency type FD model group till 6-week-old, and then modified multiple platform method （MMPM） was used for ld days. After successful modeling, they were garaged with 10 mL·kg^-1·d^-1 distilled water, 3. 125 mg·kg^-1·d^-1 Domperidone, 1. 275, 2.55, 5.1 g·kg^-1·d^-1 Pixu I recipe for 14 days. COX VA mRNA and protein expression levels in liver tissues were detected by immunohistochemistry, Western blot and Real-time PCR. Result： As compared with the normal group, the mean optical density value of COX VA proteins was increased in high-dose Pixu I recipe group （P 〈 0.05） ; COX VA protein expression levels were decreased to different degrees in Domperidone group, low-dose and medium-dose Pixu I recipe groups （P 〈 0. 01 ） , while the COX VA protein expression level in high-dose Pixu I recipe group was increased; the increase in COX VA mRNA expression was most obvious in single model group （P 〈 0.01 ）. As compared with the dual model group, COX VA protein expression levels were decreased to different degrees in Domperidone group, low-dose and medium-dose Pixu I recipe groups （P 〈 0. 01 ） , and the mRNA expression levels were also decreased while the levels in high dose group and single model group were increased without statistically significant differences. Conclusion： COX VA mRNA and protein expression levels were decreased in spleen deficiency type FD rats, and Pixu I recipe could increase the COX VA protein expression level and improve energy metabolism.