四君子汤%脾虚%疾病模型,动物%线粒体/超微结构
四君子湯%脾虛%疾病模型,動物%線粒體/超微結構
사군자탕%비허%질병모형,동물%선립체/초미결구
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3 mL/(次·只)灌胃,隔日1次,共34周.②脾虚模型组大鼠小承气汤3 mL/(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.
揹景:脾與線粒體具有相關性,中醫脾主運化不僅僅是指食物在胃腸的消化吸收,更重要的是線粒體的生物氧化產能過程.目的:分析經典健脾益氣方劑四君子湯對脾虛大鼠肝、心肌、胃黏膜和骨骼肌細胞線粒體損傷的脩複作用.設計:隨機對照觀察.單位:廣州中醫藥大學第一附屬醫院二內科和廣州中醫藥大學.材料:實驗于2004-06/12在廣州中醫藥大學第一附屬醫院內科實驗室和廣州中醫藥大學測試中心完成.選擇SD大鼠40隻,由廣州中醫藥大學動物中心提供.小承氣湯由厚樸、枳實、大黃(比例3:3:2)組成;四君子湯由黨參、白術、茯苓、甘草(比例2:2:2:1)組成,均由廣州中醫藥大學第一附屬醫院藥劑科提供,併由該院製劑室製成100%煎劑.方法:大鼠飼養1週後按隨機數字錶法分成4組,即正常對照組、脾虛模型組、自然複健組和四君子湯組.除正常對照組外,其餘3組均製作大鼠長期脾虛模型.①正常對照組大鼠常規餵養,生理鹽水3 mL/(次·隻)灌胃,隔日1次,共34週.②脾虛模型組大鼠小承氣湯3 mL/(次·隻)灌胃,隔日1次,同時隔日餵食,共34週.③自然複健組餵養方法前26週同脾虛模型組,26週後改為常規飼養,共34週.④四君子湯組餵養方法前26週同脾虛模型組;26週後改為常規飼養,同時用四君子湯灌胃8週,4mL/(次·隻),1次/d.于實驗34週末痳醉斷頭處死大鼠後,迅速取齣骨骼肌、肝髒、胃黏膜、心肌組織,用雙縮脲法測定線粒體懸液的蛋白量,併根據組織質量計算齣每剋組織的線粒體含量;應用透射電鏡觀察線粒體形態.主要觀察指標:各組大鼠的骨骼肌、肝、心肌、胃黏膜組織線粒體含量及超微形態.結果:40隻大鼠全部進入結果分析,無脫失.①實驗34週末各組大鼠骨骼肌、肝、心肌、胃黏膜組織線粒體含量比較:脾虛模型組大鼠各組織線粒體含量均顯著低于正常對照組(P＜0.01);自然複健組顯著高于脾虛模型組(P＜0.05～0.01);四君子湯組大鼠各組織線粒體含量最高,除顯著高于脾虛模型組和自然複健組外(P＜0.05～0.01),併顯著高于正常對照組(P＜0.01).②實驗34週末各組大鼠骨骼肌、肝、心肌、胃黏膜組織線粒體形態比較:脾虛模型組大鼠心肌細胞線粒體高度腫脹;肝細胞線粒體緻密質粒減少或消失,嵴斷裂;骨骼肌細胞線粒體數量減少,線粒體變小,線粒體膜結構破壞;胃壁細胞線粒體減少,線粒體內部結構不清,胃主細胞線粒體峭斷裂.自然複健組大鼠各組織的線粒體形態改變較輕,四君子湯組與正常對照組接近.結論:四君子湯具有提高脾虛大鼠骨骼肌、肝、心肌、胃黏膜組織細胞線粒體含量,脩複線粒體損傷的作用.
배경:비여선립체구유상관성,중의비주운화불부부시지식물재위장적소화흡수,경중요적시선립체적생물양화산능과정.목적:분석경전건비익기방제사군자탕대비허대서간、심기、위점막화골격기세포선립체손상적수복작용.설계:수궤대조관찰.단위:엄주중의약대학제일부속의원이내과화엄주중의약대학.재료:실험우2004-06/12재엄주중의약대학제일부속의원내과실험실화엄주중의약대학측시중심완성.선택SD대서40지,유엄주중의약대학동물중심제공.소승기탕유후박、지실、대황(비례3:3:2)조성;사군자탕유당삼、백술、복령、감초(비례2:2:2:1)조성,균유엄주중의약대학제일부속의원약제과제공,병유해원제제실제성100%전제.방법:대서사양1주후안수궤수자표법분성4조,즉정상대조조、비허모형조、자연복건조화사군자탕조.제정상대조조외,기여3조균제작대서장기비허모형.①정상대조조대서상규위양,생리염수3 mL/(차·지)관위,격일1차,공34주.②비허모형조대서소승기탕3 mL/(차·지)관위,격일1차,동시격일위식,공34주.③자연복건조위양방법전26주동비허모형조,26주후개위상규사양,공34주.④사군자탕조위양방법전26주동비허모형조;26주후개위상규사양,동시용사군자탕관위8주,4mL/(차·지),1차/d.우실험34주말마취단두처사대서후,신속취출골격기、간장、위점막、심기조직,용쌍축뇨법측정선립체현액적단백량,병근거조직질량계산출매극조직적선립체함량;응용투사전경관찰선립체형태.주요관찰지표:각조대서적골격기、간、심기、위점막조직선립체함량급초미형태.결과:40지대서전부진입결과분석,무탈실.①실험34주말각조대서골격기、간、심기、위점막조직선립체함량비교:비허모형조대서각조직선립체함량균현저저우정상대조조(P＜0.01);자연복건조현저고우비허모형조(P＜0.05～0.01);사군자탕조대서각조직선립체함량최고,제현저고우비허모형조화자연복건조외(P＜0.05～0.01),병현저고우정상대조조(P＜0.01).②실험34주말각조대서골격기、간、심기、위점막조직선립체형태비교:비허모형조대서심기세포선립체고도종창;간세포선립체치밀질립감소혹소실,척단렬;골격기세포선립체수량감소,선립체변소,선립체막결구파배;위벽세포선립체감소,선립체내부결구불청,위주세포선립체초단렬.자연복건조대서각조직적선립체형태개변교경,사군자탕조여정상대조조접근.결론:사군자탕구유제고비허대서골격기、간、심기、위점막조직세포선립체함량,수복선립체손상적작용.
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.SETTINGS: 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 and 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 officinal 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 randomlyinto 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 34 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 34th 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 I g tissue was calculated according to the tissue mass, and the mitochondrial ultrastructures 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 34th 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 ultrastructures in skeletal muscle,liver, gastric mucosa and myocardium at the end of the 34th 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.
