节点文献
基于中医“心应夏”理论的冬夏变化对血管内皮功能物质影响的实验研究
【作者】 张华；
【导师】 郭霞珍；
【作者基本信息】 北京中医药大学 ， 中医基础理论， 2010， 硕士
【摘要】 中医学“四时五脏阴阳”理论认为人与天地自然是一个统一的有机整体。自然界四时气候的变化与人体五脏功能的变化密切相关。“心应夏”是“四时五脏阴阳”理论的重要组成部分,心如何在季节的变化中保持自身稳定的生理状态,应时地适应这种变化,与冬季相比,夏季的心功能有何变化,是“心应夏”理论研究的重要内容。近年来,心血管疾病的季节节律的研究已引起学者的重视,研究季节气候变化对人体生理病理的影响对于临床治疗心血管疾病和指导人体养生具有重要的意义。1目的以“心应夏”为切入点,通过研究血管内皮功能物质的季节性变化特点,从而揭示中医“心应夏“的本质内涵,为中医“四时五脏阴阳”理论的研究提供科学依据和途径,为指导临床治疗心血管疾病提供理论依据和实验基础。2方法2.1理论研究本研究采用文献综述法和逻辑分析法,总结了从《黄帝内经》至明清的古籍以及运用现代技术对中医“心”的研究和认识,探讨“心应夏”的中医理论内涵,说明夏季脏腑的适应性调控具有一定的理论依据,进一步充实了“四时五脏阴阳”理论的内涵。2.2实验研究2.2.1实验动物：健康SD大鼠,清洁级,雄性,体重120-180g,分别于2008年12月22日(冬至)、2009年6月22日(夏至)前四十五天由北京维通利华实验动物技术有限公司购入。随机分为生理组、手术组(松果腺摘术)和伪手术组。饲养条件：自然光照,室温(冬季室温为17℃±2℃,夏季室温为25℃±2℃)。自由摄取水及饲料,饲料为普通鼠全价颗粒饲料。手术组、伪手术组分别于冬至、夏至前一个月施行手术造模,与生理组动物在相同条件下饲养到冬至、夏至当日晚8：00以后处死取材。2.2.2检测方法：血浆ET-1采用放射免疫法；血清NO采用硝酸还原酶法,血浆vWF因子采用ELISA法。2.2.3实验结果统计学分析方法：所有数据均用均数±标准差表示,运用SAS8.2统计软件分析。冬夏两季比较采用t检验(若方差不齐用t’检验),各季节组间比较采用单因素方差分析(One-Wa y Anova)检验(若方差不齐用welch检验),相关性检验采用直线相关关系。以P＜0.05为差异显著,以P＜0.01为差异极显著。3结果3.1理论研究结果本研究通过对“心应夏”理论文献的整理,揭示出中医“心应夏”的内涵,即：夏季易得心病,而冬季心病易加重。3.2实验研究结果3.2.1冬夏变化对血管内皮功能物质的影响在正常生理状态下,冬夏变化对血管内皮功能物质影响显著,表现在：(1)大鼠血浆ET-1含量冬、夏极显著差异(P<0.0001),冬季高于夏季。(2)大鼠血清NO含量冬、夏极显著差异(P<0.0001),夏季高于冬季。(3)大鼠血浆vWF含量冬、夏极显著差异(P<0.0001),冬季高于夏季。上述结果说明冬夏变化对血管内皮功能物质具有极显著的影响,其中ET-1、vWF含量冬季高于夏季,NO夏季高于冬季。3.2.2松果体——褪黑素对血管内皮功能物质冬夏变化的影响在摘除松果体的情况下,血管内皮功能物质冬夏变化节律受到影响。表现在：(1)冬季大鼠血浆ET-1含量生理组明显高于手术组(P<0.01),生理组与伪手术组之间没有差异；摘除松果体后冬、夏大鼠血浆ET-1含量无明显差异,伪手术组冬、夏大鼠血浆ET-1含量冬季明显高于夏季。(P<0.01)；夏季大鼠血浆ET-1含量生理组明显低于手术组(P<0.01),生理组与伪手术组之间没有差异；摘除松果体后冬、夏大鼠血浆ET-1含量无明显差异,伪手术组冬、夏大鼠血浆ET-1含量冬季明显高于夏季。(P<0.01)；(2)冬季大鼠血清NO含量生理组明显低于手术组(P<0.01),生理组与伪手术组之间没有差异；摘除松果体后冬、夏大鼠血清NO含量无明显差异,伪手术组冬、夏大鼠血浆NO含量夏季明显高于冬季。(P<0.0001)；夏季大鼠血清NO含量生理组明显高于手术组(P<0.0 1),生理组与伪手术组之间没有差异；摘除松果体后冬、夏大鼠血浆NO含量无明显差异,伪手术组冬、夏大鼠血浆NO含量夏季明显高于冬季。(P<0.0001)。(3)冬季大鼠血浆vWF含量生理组明显高于手术组(P<0.0001),生理组与伪手术组之间没有差异；摘除松果体后冬、夏大鼠血浆ET-1含量无明显差异,伪手术组冬、夏大鼠血浆vWF含量冬季明显高于夏季。(P<0.0001)；夏季大鼠血浆vWF含量生理组明显低于手术组(P<0.0001),生理组与伪手术组之间没有差异；摘除松果体后冬、夏大鼠血浆vWF含量无明显差异,伪手术组冬、夏大鼠血浆vWF含量冬季明显高于夏季。(P<0.0001)。上述结果说明,松果体——褪黑素在血管内皮功能物质的冬夏变化中起了调节作用,摘除松果体后,上述物质的冬夏变化差异不明显。3.2.3 ET-1／NO的冬夏变化。在生理情况下,ET-1／NO的冬夏变化极显著(P<0.0001),冬季高于夏季。均数分别为4.456±1.336和1.058±0.475。摘除松果体后,ET-1/NO的冬夏差异不明显(p>0.05)。均数分别为1.947±0.467和2.139±0.571。伪手术组的结果同生理组。上述结果说明,季节的变化会使ET-1与NO的平衡失调,在冬季,处于ET-1大于NO的状态,在夏季,ET-1与NO的水平基本一致。且这种变化也受褪黑素的影响,在摘除松果体后,这种变化不明显。3.2.4 vWF与ET-1、NO的相关性在正常生理条件下,vWF与ET-1又具有正向的线性相关关系,(p<0.01)与NO具有负向的线性相关关系(p<0.01)。冬夏的结果一致。手术组大鼠在摘除松果体后,这种线性相关关系不消失。伪手术组结果同生理组。上述结果说明,vWF的含量与ET-1正向关,与NO负相关。且这种相关性不是褪黑素对三者的共同作用造成的。4结论4.1在正常生理状态下,血管内皮功能物质呈现冬夏变化的规律。4.2血管内皮功能物质的冬夏变化与松果体的高位调节作用有关。4.3季节的变化会使ET-1与NO的平衡失调,冬季两者平衡失调,夏季两者基本平衡。4.4 vWF的含量冬夏存在极显著差异,且与ET-1正向关,与NO负相关。
【Abstract】 Theory of "Yinyang of five Zang-organs and four seasons " express that ma n is an integrated whole, and man and nature is an integrated whole. Theory of "Heart correspondence to summer "is the important content of theory of "Yinyang of five Zang-organs and four seasons ". The researches on theory of "Heart correspondence to summer " mainly focus on how heart adapt the changes of nature to mainta in the balance of physiological function.In recent years, the research of seasonal rhythm of cardiovascular disease has attracted attention of scholars. Studying on the seasonal climate influencing on physiological function is of great importance to clinical diagnosis, theory and preventing diseases.1. ObjectiveTaking the idea of "Heart correspondence to summer" in TCM as a breaking point,we research on the seasonal changes of vascular endothelial function materials to discuss the truth of theory of "Yinyang of five Zang-organs and four seasons ", which can enrich the scientific methods of theory of "Yinyang of five Zang-organs and four seasons”.Through this research,we can give some theoretic and experimental proofs on clinical diagnosis, theory and preventing diseases.2. Methods2.1 Theoretical methodsThe methods of the documentary substances and logical analysis were used to explain and analyze the physiological mechanism of "Heart correspondence to summer”.It showed that "Zang Fu proper to nature " had theoretical basis, and enriched the theoretical connotation of "Yinyang of five Zang-organs and four seasons ".2.2Experimental methods2.2.1 Experimental animalsTwenty-four SD male rats, weighted 120-180g,brought from Beijing WeitongLihua animal study center forty-five days before winter solstice and summer solstice were randomly divided into three groups:normal group, operation group(pinea lectomy group) and fake-operation group. All rats received na tural light, ate and drank freely, and were controlled under certa in room tempera ture was 17℃±2℃in winter, and it was 25℃±2℃in summer. Operation group and fake-operation group are operated a month before winter solstice and summer solstice.All the rats would be raised till winter solstice and summer solstice, on these days at 8 o’clock at night, they would be beheaded.2.2.2 Test methodsTo assay the ET-1 in plasma, Radio-immunity was engaged. To assay the contents of NO in serum, Nitrate reductase was engaged. To assay the contents of vWF in plasma, ELISA was engaged.2.2.3 Analysis method:All the data were showed by mean±standard error. The differences of groups were tested by SAS8.2. The values were calculated with Dunca n’s multiple range test or Student’s test. P< 0.05 meant obvious difference.3Results3.1 Results of theoretical study After analysis of the literature of the theory of "Heart correspondence to summer ",we think the meaning of this theory is:summer is the season which easy to get cardiovascular disease, winter is the season which cardiovascular disease easy to deteriorate.3.2 Results of experiments3.2.1 Seasonal change can influence In normal stasis, the vascular endothelial function substances have the character of seasonal changes.(1) ET-1 in plasma has significant difference between winter and summer (P＜0.0001). It was higher in winter and lower in summer.(2)NO in serum has significant difference between winter and summer (P＜0.0001). It was higher in summer and lower in winter.(3)vWF in plasma has significant difference between winter and summer (P＜0.0001). It was higher in winter and lower in summer.These results indicate that winter and summer changs has significant impact on vascular endothelial function substances. ET-1、vWF is higher in winter than in summer, NO is higher in summer than in winter.3.2.2 Pineal gland-Melatonin can influence on the seasonal changes of vascular endothelial function substances.If we remove the Pineal gland, the seasonal changes of vascular endothelial function substances can be destroyed:(1) In winter,normal group of ET-1 in plasma was significant higher than operation group of it (P＜0.01). There is no difference between normal group and fake-operation group. After removed the Pineal gland, There is no difference of ET-1 in plasma between winter and summer. Fake-operation group of ET-1 in plasma was significant higher in winter than in summer (P<0.01). In summer,normal group of ET-1 in plasma was significant lower than operation group of it (P＜0.01). There is no difference between normal group and fake-operation group. After removed the Pineal gland, There is no difference of ET-1 in plasma between winter and summer. Fake-operation group of ET-1 in plasma was significant higher in winter than in summer (P＜0.01)(2)In winter,normal group of NO in serum was significant lower than operation group of it (P＜0.01). There is no difference between normal group and fake-operation group. After removed the Pineal gland, There is no difference of NO in serum between winter and summer. Fake-operation group of NO in serum was significant higher in summer than in winter (P＜0.0001). In summer,normal group of NO in serum was significant higher than operation group of it (P＜0.01). There is no difference between normal group and fake-operation group. After removed the Pineal gland, There is no difference of NO in serum between winter and summer. Fake-operation group of NO in serum was significant higher in summer than in winter (P＜0.0001)(3)In winter,normal group of vWF in plasma was significant higher than operation group of it(P＜0.0001). There is no difference between normal group and fake-operation group. After removed the Pineal gland, There is no difference of vWF in plasma between winter and summer. Fake-operation group of vWF in plasma was significant higher in winter than in summer (P<0.0001).In summer,normal group of vWF in plasma was significant lower than operation group of it (P＜0.0001). There is no difference between normal group and fake-operation group. After remove the Pineal gland, there is no difference of vWF in plasma between winter and summer. Fake-operation group of vWF in plasma was significant higher in winter than in summer (P<0.0001)These results indicate that Pineal gland-Melatonin can influence on the seasonal changes of vascular endothelial function substances. If we remove the Pineal gland, there is no difference of these substances between winter and summer.3.2.3seasonal changes of ET-1/NOIn normal stasis, ET-1/NO has significant difference between winter and summer (P＜0.0001). It was higher in winter and lower in summer. The means are 3.983±0.611 and 1.044±0.406 respectively.After removed the Pineal gland, There is no difference of ET-1/NO between winter and summer(P<0.0001). The means are 2.056±0.237 and 2.044±0.438 respectively. The results of fake-operation group are similar as normal group.These results indicate that seasonal changes can imbalance the contents of ET-1 and NO. In winter ET-1 is higher than NO and in summer ET-1 and NO are almost equal. What’s more, these changes are influenced to the Melatonin. If we remove the Pineal gland, these changes are not obviously.3.2.4Correlation between vWF and ET-1、NOIn normal stasis, it has positively correlation between vWF and ET-l,(P<0.01) and negative correlation between vWF and NO(P<0.01). The results of winter and summer are similar.If we remove the Pineal gland, these correlations not disappear.The results of fake-operation group are similar as normal group.These results indicate that it has positively correlation between vWF and ET-1 and negative correlation between vWF and NO. These correlations are not the result of melatonin.4 Conclusion4.1 In normal stasis, the vascular endothelial function substances have the character of seasonal changes.4.2 The seasonal changes of vascular endothelial function substances are influence to Pineal gland.4.3 Seasonal changes can imbalance the contents of ET-1 and NO. In winter ET-1 is higher than NO and in summer ET-1 and NO are almost equal.4.4 It has positively correlation between vWF and ET-1 and negative correlation between vWF and NO.