摘要
背景:脾与线粒体具有相关性,中医脾主运化不仅仅是指食物在胃肠的消化吸收,更重要的是线粒体的生物氧化产能过程。目的:分析经典健脾益气方剂四君子汤对脾虚大鼠肝、心肌、胃黏膜和骨骼肌细胞线粒体损伤的修复作用。设计:随机对照观察。单位:广州中医药大学第一附属医院二内科和广州中医药大学。材料:实验于2004-06/12在广州中医药大学第一附属医院内科实验室和广州中医药大学测试中心完成。选择SD大鼠40只,由广州中医药大学动物中心提供。小承气汤由厚朴、枳实、大黄(比例3∶3∶2)组成;四君子汤由党参、白术、茯苓、甘草(比例2∶2∶2∶1)组成,均由广州中医药大学第一附属医院药剂科提供,并由该院制剂室制成100%煎剂。方法:大鼠饲养1周后按随机数字表法分成4组,即正常对照组、脾虚模型组、自然复健组和四君子汤组。除正常对照组外,其余3组均制作大鼠长期脾虚模型。①正常对照组大鼠常规喂养,生理盐水3mL/(次·只)灌胃,隔日1次,共34周。②脾虚模型组大鼠小承气汤3mL/(次·只)灌胃,隔日1次,同时隔日喂食,共34周。③自然复健组喂养方法前26周同脾虚模型组,26周后改为常规饲养,共34周。④四君子汤组喂养方法前26周同脾虚模型组;26周后改为常规饲养,同时用四君子汤灌胃8周,4mL/(次·只),1次/d。于实验34周末麻醉断头处死大鼠后,迅速取出骨骼肌、肝脏、胃黏膜、心肌组织,用双缩脲法测定线粒体悬液的蛋白量,并根据组织质量计算出每克组织的线粒体含量;应用透射电镜观察线粒体形态。主要观察指标:各组大鼠的骨骼肌、肝、心肌、胃黏膜组织线粒体含量及超微形态。结果:40只大鼠全部进入结果分析,无脱失。①实验34周末各组大鼠骨骼肌、肝、心肌、胃黏膜组织线粒体含量比较:脾虚模型组大鼠各组织线粒体含量均显著低于正常对照组(P<0.01);自然复健组显著高于脾虚模型组(P<0.05～0.01);四君子汤组大鼠各组织线粒体含量最高,除显著高于脾虚模型组和自然复健组外(P<0.05～0.01),并显著高于正常对照组(P<0.01)。②实验34周末各组大鼠骨骼肌、肝、心肌、胃黏膜组织线粒体形态比较:脾虚模型组大鼠心肌细胞线粒体高度肿胀;肝细胞线粒体致密质粒减少或消失,嵴断裂;骨骼肌细胞线粒体数量减少,线粒体变小,线粒体膜结构破坏;胃壁细胞线粒体减少,线粒体内部结构不清,胃主细胞线粒体峭断裂。自然复健组大鼠各组织的线粒体形态改变较轻,四君子汤组与正常对照组接近。结论:四君子汤具有提高脾虚大鼠骨骼肌、肝、心肌、胃黏膜组织细胞线粒体含量,修复线粒体损伤的作用。
BACKGROUND： Spleen is correlated with mitochondrion. The “spleen governs movement and transformation of food and liquids” in traditional Chinese medicine refers to not only the digestion and absorption of food in gastrointestinal tract, but also the process of biological oxidation and energy production of mitochondrion. OBJECTIVE： To analyze the effects of Si-Jun-Zi decoction （SJZD）, a typical prescription for invigorating spleen and replenishing qi, on repairing the mitochondrial damage of cells of liver, myocardium, gastric mucosa and skeletal muscle in rats with spleen asthenia.
DESIGN： A randomized controlled observation.
SETIINGS： Second Department of Internal Medicine, the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine; Guangzhou University of Traditional Chinese Medicine. MATERIALS： The experiments were carried out in the internal medicine laboratory, the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine a0d the testing center of Guangzhou University of Traditional Chinese Medicine from June to December in 2004. Forty SD rats were provided by the animal center of Guangzhou University of Traditional Chinese Medicine. Xiao Cheng-Qi decoction consisted of officinai magnolia bark, immature bitter orange and rhubarb according to the ratio of 3：3：2; SJZD consisted of radix codonopsitis pilosulae, largehead atractylodes rhizome, India bread and liquorice root according to the ratio of 2：2： 2：1, and it was provided by the Department of Pharmacy, the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, and made into 100% decoction. METHODS： After raised for 1 week, the SD rats were divided randomly into 4 groups： normal control group, spleen asthenia group, natural convalescence group and SJZD-treated group, and the rats in the latter three groups were made into models of long-term spleen asthenia. Rats in the normal control group were fed with normal chow, intragastric administered with saline （3 mL）, once every other day for 3d weeks. Rats in the spleen asthenia group were intragastricly administered with Xiao Cheng-Qi decoction （3 mL） and fed every other day for 34 weeks, the rats in the natural convalescence group were fed with normal chow for 8 weeks after being treated as those in the spleen asthenia group for 26 weeks, and those in the SJZD-treated group were treated as those in the spleen asthenia group for 26 weeks, and then intragastricly administered with SJZD （4 mL, once a day） and fed with normal chow for 8 weeks. At the end of the 34^th week, the rats were decapitated under anesthesia, and the skeletal muscle, liver, gastric mucosa and myocardium were taken out rapidly. The protein amounts in mitochondrial suspension were detected with the biuret method, the content of mitochondria in 1 g tissue was calculated according to the tissue mass, and the mitochondrial uhrastructures were observed under transmission electron microscope.
MAIN OUTCOME MEASURES： The contents and ultrastructures of mitochondria in skeletal muscle, liver, gastric mucosa and myocardium were observed.
RESULTS： All the 40 rats were involved in the analysis of results without deletion. ① At the end of the 34^th week, the mitochondrial contents of skeletal muscle, liver, myocardium and gastric mucosa were all significantly lower in the spleen asthenia group than in the normal control group （P 〈 0.01）, and markedly higher in the natural convalescence group than in the spleen asthenia group （P 〈 0.05-0.01）. The mitochondrial contents of the tissues were the highest in the SJZD-treated group, which were significantly higher than those in the spleen asthenia group and natural convalescence group （P 〈 0.05-0.01）, as well as the normal control group （P 〈 0.01）. ② Comparison of mitochondrial ultrastruetures in skeletal muscle, liver, gastric mucosa and myocardium at the end of the 34^th week： In the spleen asthenia group, the mitochondria of myocardial cells were seriously swollen, the compact substance of hepatocellular mitochondria were decreased or disappeared and the crest disrupted; the mitochondria of skeletal muscle were shrunk and decreased, mitochondrial membranes were disorganized and crest disappeared, mitochondrial membranes were disorganized and crest disappeared; For gastric parietal cell of spleen asthenia, the amount of mitochondria reduced, inner structure confused, mitochondrial crestae of gastric chief cell was broken. In the natural convalescence group, the changes of the mitochondrial morphology were slight. The mitochondrial morphology in the SJZD group was close to those in the normal control group.
CONCLUSION： SJZD has the effects of increasing the contents of mitochondria in skeletal muscle, liver, myocardium and gastric mucosa and repairing the damaged structure of mitochondria because of spleen asthenia.
出处
《中国临床康复》
CSCD
北大核心
2006年第39期170-173,F0003,共5页
Chinese Journal of Clinical Rehabilitation
基金
国家自然科学基金资助项目(30371706)~~
