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目的观察脾虚模型大鼠骨骼肌线粒体SDH活性、骨骼肌PGC-1αmRNA及蛋白表达变化及四君子汤对其影响,探讨脾虚证能量代谢障碍机制。方法采用皮下注射利血平(0.5mg·kg~(-1)·d~(-1))方法建立脾虚大鼠模型,造模成功后,将模型组大鼠分为四君子汤组和脾虚组,每组各7只,另设正常组7只。四君子汤给药组灌胃四君子汤10g·10ml~(-1)·kg~(-1),脾虚组和正常组灌胃等量的生理盐水,每日1次,持续3周,记录大鼠的一般状况,3周后检测大鼠尿D-木糖排泄率,提取骨骼肌线粒体,检测大鼠骨骼肌线粒体SDH活性、骨骼肌PGC-1αmRNA及蛋白的表达情况。结果与正常组相比,脾虚组大鼠骨骼肌线粒体SDH活性显著降低(P<0.01);大鼠骨骼肌PGC-1αmRNA及蛋白表达显著降低(P<0.01);与脾虚组相比,四君子汤组大鼠骨骼肌线粒体SDH活性升高(P<0.05),大鼠骨骼肌PGC-1αmRNA及蛋白表达升高(P<0.05)。结论脾虚模型大鼠骨骼肌线粒体存在功能损伤,呼吸链酶活性降低,导致线粒体氧化磷酸化下降,机体生物合成、能量代谢障碍;健脾益气中药可提高线粒体氧化磷酸化水平,改善能量代谢。
Objective To observe the changes of mitochondrial SDH activity, skeletal muscle PGC-1αmRNA and protein expression in skeletal muscle of rats with spleen-deficiency syndrome and the effect of Sijunzi Decoction on it, and to explore the mechanism of energy metabolism disorders in spleen deficiency syndrome. Methods Subcutaneous injection of reserpine (0.5mg · kg -1 · d -1) was used to establish the model of splenic asthenia in rats. After successful model establishment, the model rats were divided into Sijunzi Decoction group and spleen deficiency group , Each group of 7, another set of normal group of 7. The Sijunzi decoction group was fed with Sijunzi Decoction 10g · 10ml -1 (-1) · kg -1, and the rats in the spleen deficiency group and the normal group were given the same amount of normal saline by gavage once a day for 3 weeks. After 3 weeks, D-xylose excretion rate of rat urine was measured. Skeletal muscle mitochondria were extracted and mitochondrial SDH activity and skeletal muscle PGC-1α mRNA and protein expression were detected. Results Compared with the normal group, the activity of SDH in skeletal muscle of spleen deficiency group was significantly decreased (P <0.01), while the PGC-1α mRNA and protein expression in skeletal muscle was significantly decreased (P <0.01). Compared with the spleen deficiency group, The SDH activity of mitochondria in skeletal muscle of rats increased (P <0.05), and the mRNA and protein expression of PGC-1α in skeletal muscle of rats increased (P <0.05). Conclusion Spleen deficiency model rats have functional impairment of skeletal muscle mitochondria and decreased respiratory chain enzyme activity, resulting in decreased mitochondrial oxidative phosphorylation, body biosynthesis and energy metabolism disorders. Chinese herbal medicine Jianpi Yiqi can increase mitochondrial oxidative phosphorylation and improve energy metabolism.