“肺应秋”调控机制的理论整理和免疫学实验研究

【作者】 吴同玉；

【导师】 刘燕池；

【作者基本信息】 北京中医药大学 ， 中医基础理论， 2005， 博士

【摘要】 中医藏象学说是中医学理论体系的核心组成部分,《内经》中五脏的功能,着重在于其调控功能,其调控功能不仅是对机体本身进行调节,还对自然界的变化起着协调共振的适应性调节作用。“四时五脏相应”(简称“时脏相应”)是《黄帝内经》中一个重要理论观点,“时脏相应”理论主要是说人体内在的脏腑生理机能与外界时辰季节变化具有同步的相应性变化规律。 1.目的 本课题以“肺应秋”为切入点,从免疫学实验角度探讨中医肺藏的生理调控机制,通过“肺应秋”调控机制与免疫物质相关性的实验研究,进一步实证脏腑应时适应性调控的科学性和可重复性,为中医藏象学说的研究开辟新的途径。 2.方法 2.1理论研究:采用文献学、逻辑分析等方法,对中医“天人相应”,脏腑应时适应性调控的内涵进行探讨分析,并以“肺应秋”为切入点,提出了从免疫学角度进行实验研究的方法和途径。 2.2实验研究 2.2.1实验动物:SD雄性大鼠,24只,体重180-220g,3月龄,分别在春分和秋分前四十天由北京维通利华动物研究中心购入。随机分为正常组、伪手术组和松果腺切除模型组,每组各8只。饲养条件:自然光照,室温。自由摄取水及饲料,饲料为普通鼠全价颗粒饲料。手术组和伪手术组分别于二分前一个月实行松果腺切除术和伪手术,与正常组动物在相同条件下饲养到二分日晚上8:00以后进行实验。 2.2.2实验指标检测方法:血清IgG采用单向免疫扩散法,血清IL-2采用ELISA法,脾脏T淋巴细胞转化率采用MTT法,肺泡灌洗液中巨噬细胞吞噬率采用鸡红细胞吞噬法,肺泡灌洗液中IFN-γ和IL-6采用ELISA法,血清和肺组织的MT以及肺组织的IL-10均采用放射免疫分析法。 2.2.3实验结果统计分析方法:所有数据以(?)±s表示,组间差异利用SPSS10.0统计软件包进行ANOVA检验,以P<0.05差异有显著性。 3.结果 3.1春季和秋季正常组大鼠的机体和肺的各项免疫指标,包括血清和肺组织中褪黑素含量的变化,均呈现出春高秋低的分布趋势,其中脾脏指数、胸腺指数、脾T淋巴细胞转化率以及AM吞噬率变化非常明显,p<0.01;BALF中IL-6亦有明显的变化,差异具有显著性,p<0.05。 3.2切除松果腺后,以上各项指标在春秋季节的两组大鼠之间无明显差异。同时亦发现切除松果腺后,春分和秋分时大鼠的免疫指标和褪黑素均呈现出相对于正常组有增高的趋势,其中脾T淋巴细胞转化率、血清IgG和AM吞噬率增高非常明显,差异具有非常显著性,p<0.01;秋分时手术模型组大鼠BALF中IFN-γ亦明显高于正常对照组,p<0.05。更多还原

【Abstract】 The theory of Traditional Chinese Medicine(TCM) Zangxiang is the core of TCM theory system.The TCM five visceras including form visceras and spirit visceras, on the basis of form visceras, we should know clearly the spirit visceras, namely the functional visceras.The theory of the correspondence between Five Zang and time is an important point in <Neijing>.The theory of correspondence between time and Zang means the corresponding change of the function of the body along with the change of environment. The five zang-organ is the modulating system of the human body.The action of five zang-organ in <Neijing> is the modulating function. Five zang-organ can not only modulate the body itself, but also modulate its correspondence with the nature.1 .ObjectiveOn the basis of "lung corresponding to autumn", we discuss the modulation mechanism of lung from the point of immunity, and the relative study of lung and immunity.We want to prove that the modulation of zang and fu-organs corresponding to time is scientific and can be repeated.2.Method2.1 Twenty-four SD male rats, 180-220g weight, three months age.We got it from Beijing Weitong Lihua animal study center forty days before vernal equinox and autumnal equinox, then divided into three groups randomly:normal group, fake group ans pinealectomy group.They were raised to take water and foods freely.Fake group and Px group were operated a month before vernal equinox and autumnal equinox, then on 8:00pm of vernal equinox and autumnal equinox, we should kill all the rats.2.2 Serum IgG was detected in the way of unilateralism immunity diffusion.Serum IL-2 was detected in the way of ezyme-linked immunosorbent assay.T lymphocyte translation rate of spleen was detected in MTT.IFN-yand IL-6 in bronchoalveolar lavage fluid were detected in ELISA and IL-10 and MT were detected in the way of radiation immunity analysis,etc.2.3 Data analysis method:All the data were showed in the form of x±s.The difference ofgroup was tested in SPSS 10.0.3.ResultsAll the substances including the immune materials and melatonin in normal group were low in autumn and high in spring, and index of spleen and thymus, T lymphocyte translation rate of spleen and IL-6 in BALF, etc.changed much.The difference is clear, p< 0.05.While pinealectomy, all the data in rats between spring and autumn didn’t change much, and the substances all showed the trends of ascending, and some indexes change much compare to normal group.4.Conclusions(l)Pineal gland is one of the important medium in modulating mechanism of "lung correspondence with autumn".The results proved that melatonin in serum and lung is higher in spring and lower in autumn at normal state. When removed the pineal gland, the contents of melatonin in lung and serum increased as feedback, it has a clear difference compared with the normal group. The rhythm of melatonin higher in spring and lower in autumn is not evident while pinealectomy.The production of melatonin in serum and lung is not in the pineal gland while pinealectomy, it may be excreted from other organs, but the results connect with the stimulation of pinealectomy on neuroendocrine.We also found that the exponents of thymus gland and spleen, spleen T lymphocyte translation rate, IgG and IL-2 in serum, function ofmacrophage in pulmonary alveoli, IFN-Y and IL-6 in pulmonary alveoli fluid, IL-10 in lung tissue, etc. all showed the trends of higher in spring and lower in autumn in normal group, resembling the trends of melatonin in serum and lung tissue.Melatonin in serum has negative correlation with IgG in Spring Equinox, and melatonin in lung has a direct correlation with the function ofmacrophage in pulmonary alveoli in Autumn Equinox, which may give the reasons that rhythms of immunity in body and lung have an evident relationship with the modulation of melatonin in pineal gland. So we can say that pineal gland may be one of important medium in modulating the relationship between season and zang-fu organs.We affirmed that melatonin modulate the immunity of lung.(2)The theory of "lung correspondenc更多还原