Discussion On The Scientific Connotation Of The Theory Of "Four - Time - Five - Body - Yin - Yang" From The Seasonal Changes Of Hypothalamus - Pituitary - Gonadal Axis Function

"Four Seasons-five organs-Yin and Yang" theory from the "Yellow Emperor", is in the "Heaven corresponding" guiding ideology formation.The theory emphasizes a variation of adaptive resonance coordination between the physiology of the human body internal organs and the outside natural hour and seasons,becoming an important starting point to research the viscera state’s essence from a angle of time medical.1 ObjectiveAccording to experimental study of correlation between melatonin and the hypothalamus-pituitary-gonadal axis(HPGA) under normal seasonal physiological state, investigating the seasonal impact on the function of HPGA and it’s pathway,also melatonin’s season-related effects to HPGA,from different levels of melatonin, hormone of HPGA, melatonin receptor level, the rate-limiting enzyme of melatonin synthesis activity and intracellular signal transduction.To confirmed the body’s internal functions of time variations has synchronization with outside natural seasons from the physical point. And to provide a scientific basis for the theory connotation of "Four Seasons-five organs-Yin and Yang" theory.2 Research Methods2.1 Literature ReviewUsing the methods of philology and logic analysis to comb seasonal change related to human organs’function records in ancient Chinese medicine books, and analyzing it from points of physiological changes,diagnosis. treatment. prevention and care, to summarize literature basis of the modern research about four seasons factors affecting the function of body organs.2.2 Experimental Research2.2.1 Research IdeasFrom the start of correlation between melatonin and HPGA. In the spring equinox, summer solstice, autumnal equinox and winter solstice.Detecting melatonin and gonadal hormones levels in plasma, melatonin and melatonin receptors levels in hypothalamus, pituitary and testis,also they secreted hormones content,to observe the seasonal changes impact on HPGA function. Detecting the seasonal variations of AANAT、HIOMT activity in the hypothalamus, pituitary gland, and gonad, and analyzing seasonal variation of specific substances features that make melatonin carry out seasonal regulation of HPGA from rhythm of melatonin synthesis rate-limiting enzyme. Finally detecting the third messenger c-fos and c-jun’s expression intensity of seasons in the hypothalamus, pituitary and testis,to explore the variations of HPGA function in view of intracellular signal transduction.2.2.2 Experimental AnimalsSPF male Syrian golden hamsters, two months old,120-140g/one. Respectively, purchased from the Beijing Weitonglihua Animal Research Center before two weeks of the spring equinox, summer solstice, autumnal equinox and winter solstice, divided into three groups of eight every season, a week adaptive feeding. Rearing conditions:natural light, humidity 50%-70%, the winter temperature 18±2 ℃, summer 25±2℃. Normal mice feed for Beijing Vital River Laboratory Animal Center of the full price of pellets. Respectively, in the spring equinox, summer solstice, autumnal equinox and winter solstice, taking experiment under the 5w red lamp in a dark room at 22:00.2.2.3 DetectionPlasma MT, GnRH, FSH, LH, T and the hypothalamus, pituitary, testis MT, MR, GnRH, FSH, LH, T,AANAT, HIOMT detected by enzyme-linked immunosorbent assaymethod (ELISA); c-fos mRNA and c-jun mRNA expression in the hypothalamus, pituitary and testis using in situ hybridization assay.2.2.4 Analysis of statistical methods(1) Image analysis of in situ hybridization using a professional image analysis software Image-Pro Plus;(2) Each set of data were expressed as mean±standard deviation (x±s) that analysed by SPSS 20.0 statistical software, P<0.05 was considered statistically significant.3 Results3.1 The results of literature reviewCombing through books, in the " Yellow Emperor",there are many articles that elaborate seasonal changes associated with the function of organs, done a lot of discussion which combined with the clinical diagnosis, treatment, and disease prevention, which lay the foundation for the development of the theory of traditional Chinese medicine and clinical diagnostic, and for the research of internal organs function through observing physiological and pathological changes of organs basing on seasonal changes. After that, many physicians put forward their views on the seasons effects on diagnosis and treatment of disease. All of these have provided a lot of literature basis for the modern study of the essence of viscera state from the perspective of time.3.2 Experimental Results3.2.1 The Seasonal Changes of melatonin and HPGA HormonesUnder normal physiological conditions, plasma and the content of MT, MR, GnRH, FSH, LH, T in gonadal axis tissues have shown changes in seasonal trends (p<0.05). The content of plasma MT, the MT and MR in hypothalamus, the MT in testis are higher in the spring and winter than in summer and autumn; the MT in pituitary tissue are high in spring and summer, low in autumn and spring, and the MR in summer and autumn are higher than in spring and winter, but the MR in testis in spring and autumn are higher than in summer and winter. The Plasma GnRH and T in autumn and winter are higher than in spring and summer; the FSH, LH in spring and autumn are higher than in summer and winter. The hypothalamus GnRH, the LH, FSH in pituitary tissue, the T in testicular tissue generally show a decreasing trend from the vernal equinox to the winter solstice. The relative analyses show that there is a correlation between the MT, MR in each tissue of gonadal axis and the hormone in the tissue. The experimental results show that the function of the gonadal axis has the characteristics of changing with seasons, and the melatonin may play a regulatory role in this process.3.2.2 The seasonal changes of the activity of AANAT and HIOMT in each tissue of HPGAUnder normal physiological conditions, the hypothalamus, pituitary, the activity of AANAT, HIOMT in testicular tissue have shown changes in seasonal trends (p<0.05), which is consistent with the trend of the MT in the same tissue. The activity of the AANAT, HIOMT in hypothalamus and testis have a high performance both in spring and wither, but low in summer and autumn; the AANAT, HIOMT in pituitary tissue are high in spring and summer, but low in autumn and winter. The relative analyses show that the MT in hypothalamus, pituitary and testis has a positive correlation with the activity of the AANAT. HIOMT in the same tissue (p<0.05). synchronizing change with seasonal variations. The experimental results show that the tissue of gonadal Axis are able to synthesize a small amount of MT, which is under the influence of the seasonal rhythm of the activity of AANAT. HIOMT.3.2.3 The seasonal expression of c-fos and c-jun mRNA in each tissues of HPGAUnder normal physiological conditions, the expression of hypothalamus, the c-fos mRNA and c-jun mRNA in pituitary tissue, the c-jun mRNA in testis exhibit the characteristics of the seasons change (p<0.05). The c-fos mRNA in the hypothalamus tissues expresses strongly in spring and summer than in autumn and winter, and in the pituitary gland and testis expresses strongly in autumn and winter than in spring and summer, but the different expression in the testis was not statistically significant (P> 0.05). The expression of the c-jun mRNA in the hypothalamus, pituitary, testis has shown a strong tendency in autumn and winter than in spring and summer. in the pituitary and testis, trend of c-fos and c-jun’s seasonal expression are similar to seasonal trends of plasma GnRH, and T content.All of these show that, seasonal changes of HPGA function relate with seasonal expression of c-fos and c-jun in hypothalamus, pituitary and testis4 Conclusion4.1 Under physiological conditions, HPGA function has the characteristics of changing form seasons, and the melatonin may play a regulatory role in this process.4.2 A small number of melatonin can be synthesized in each tissue of HPGA, which present the characteristics of seasonal changes influenced by the seasonal rhythm of the rate-limiting enzyme of melatonin synthesis.4.3 Seasonal changes of HPGA function relate with seasonal variations of intracellular signal transduction substances.4.4 Conclusion above provides an experimental basis for"Four Seasons-five organs-Yin and Yang" theory.