脾气虚证和脾不统血证模型大鼠脾主肌肉实质的探讨

Discussion on the Essence of Pi Dominating Muscle of Pi-qi Deficiency Syndrome and Pi Failing to Control Blood Syndrome Rat Model

目的研究脾气虚证和脾不统血证模型大鼠骨骼肌组织线粒体能量代谢变化,探讨中医脾主肌肉理论的内涵实质。方法将48只SD雄性大鼠按随机数表法分为正常对照组、低分子肝素钠组、脾气虚组和脾不统血组,每组12只。复制脾气虚证和脾不统血证大鼠模型,测定各组大鼠骨骼肌ATP含量、线粒体呼吸链复合物(ComplexⅠ、ComplexⅡ、ComplexⅢ、ComplexⅣ)活性、柠檬酸合酶(CS)活性、mt DNA拷贝数、COXⅣ蛋白和Cyt-C蛋白及其mRNA表达量变化。结果与正常对照组比较,中期和末期脾气虚组和脾不统血组大鼠骨骼肌的ATP含量、线粒体复合物活性、CS活性、mt DNA拷贝数、COXⅣ蛋白及其mRNA表达量均降低,Cyt-C蛋白及其mRNA表达量均升高(P <0. 05,P <0. 01);与脾气虚组比较,中期和末期脾不统血组大鼠骨骼肌ATP含量、CS活性均降低(均P <0. 05);中期脾不统血组大鼠骨骼肌ComplexⅠ、ComplexⅡ、ComplexⅢ活性降低,末期脾不统血组大鼠骨骼肌ComplexⅠ、ComplexⅢ、ComplexⅣ活性降低(P <0. 05,P <0. 01);中期脾不统血组大鼠骨骼肌mt DNA拷贝数升高(P <0. 05);脾不统血组大鼠骨骼肌COXⅣ、Cyt-C蛋白及其mRNA表达量比较,差异无统计学意义(均P> 0. 05)。结论脾气虚证大鼠和脾不统血证大鼠骨骼肌线粒体能量代谢障碍存在多方面的影响变化,且脾气虚证发展成为脾不统血证的过程中,大鼠骨骼肌能量代谢障碍逐渐加重。

Objective To study the changes of mitochondrial energy metabolism in skeletal muscle tissue of rats with Pi-qi deficiency syndrome and Pi failing to control blood syndrome,and to explore the connotation and essence of theory which was the Pi nourishing the muscles. Methods Totally 48 SD male rats were divided into normal control group,low molecular weight heparin sodium group,Pi-qi deficiency group and Pi failing to control blood syndrome group according to the random number table,12 in each group. The rat model of Pi-qi deficiency syndrome and Pi failing to control blood syndrome were established. The ATP content,mitochondrial respiratory chain complex( Complex Ⅰ,Complex Ⅱ,Complex Ⅲ,ComplexⅣ) activity,citrate synthase( CS) activity,mt DNA copy number,the expression of COX Ⅳ and Cyt-C and their mRNA were measured in the skeletal muscle of the rats. Results Compared with the normal control group,ATP content,mitochondrial complex activity,CS activity,mt DNA copy number,COX Ⅳ protein and mRNA expression of skeletal muscle in the Piqi deficiency group and Pi failing to control blood group were decreased in the middle and end stage,while the expression levels of Cyt-C protein and mRNA in above groups were increased in the middle and end stage(P < 0. 05,P < 0. 01). Compared with the Pi-qi deficiency group,the ATP content and CS activity of skeletal muscle in the Pi failing to control blood group were decreased( all P < 0. 05). The activity of Complex Ⅰ,Complex Ⅱ and ComplexⅢ in skeletal muscle of rats in the Pi failing to control blood group was decreased in the middle stage,while the activity of Complex Ⅰ,Complex Ⅲ and ComplexⅣ in skeletal muscle of rats Pi dysplasia was decreased in the end stage(P <0. 05,P <0. 01). The mt DNA copy number of skeletal muscle was increased in the above group(P < 0. 05). There was no significant difference in the expression of COXⅣ,Cyt-C protein and mRNA between the Pi failing to control blood group and Pi-qi deficiency group( all P >0. 05). Conclusions There were many changes in skeletal muscle mitochondrial energy metabolism disorder in rats with Pi-qi deficiency syndrome and Pi failing to control blood syndrome. And the skeletal muscle energy metabolism disorder gradually increased in the process of Pi-qi deficiency syndrome.