Theory And Experimental Study Of Exogenous Cold-evil Injury Lung

The cold-evil, one of the six evils, is the common pathogenic factor of diseases. Which from outside was named exogenous cold-evil. Exogenous cold-evil can impair lung easily. There are many famous ideas in " Huangdi Neijing", such as "Cold body and cold drink injure lung ","lung aversion to cold". The research of Cold syndrome is sufficient, But the cold-evil theory and empirical research was obviously insufficient for20years. Through guided by the etiological factor and pathogenesis theory of traditional chinese medicine and reviewed the documents, the topic is established the animal model of exogenous cold-evil injure lung and examine the antibody, AQP-1, BAFF and NF-κB expression of lung which reflect the governing regulation of water passages and defending function. The topic provides the evidences to explain the machinisms of exogenous cold-evil, enrich the theories of six evils and give new index of the treatment based on syndrome differentiation for exogenous cold-evil. The research have the innovation in the theory and give guidance to the clinics for exogenous cold-evil.1. Theory researchCold is one of the six influences of the natural environment and is a normal climatic variation of nature. it is the primary climatic factors in winter. when the climatic variations are too unusual, the occurrence of the natural cold is so excessive that people can not adapt to it, such cold will affect the human body and cause diseases. The cold under such circumstances is then called "cold-evil". Cold-evil in the traditional chinese medicine not only includes to temperature in environment, but also relates to meteorological factors, biological factors, chemical factors, body constitution and responsivity, etc.Lung locates in thoracic cavity, so each one, which is the organ’s canopy and connecting all vessels. The lung governs qi, performs respiration, governs disseminating, cleansing and descending, and regulates the waterways. Lung can diffuse defend qi on the skin and hair to achieve the function of resisting the evils outside.There are many theories in " Huangdi Neijing", such as "Cold body and cold drink injure lung ","lung aversion to cold". Aversion to cold is closely related physiological characteristics of the lung. Lungs connected directly through the nasal passage with the outside, and delicate which called "fragile organ" in the traditional chinese medicine. The lung governs the skin and hair, when the skin and hair catch cold, it can also involve the lung.The basic pathogenesis of exogenous cold-evil injury lung is lung yang damage. Lung yang is warm, promoting, diffusing part of lung qi. Cold evil is yin pathogen and apt to attack yang. Exogenous cold-evil attacks lung and damages the lung yang, which leads failure of lung qi in dispersion and regulates the waterways, defend qi can not diffuse on the skin and hair and failure to resisting the evils outside.2. Experiment study2.1Animals and grouping50specific pathogen free Kunming mice,17±g, male and female in half, have been conventionally breeded in the laboratory for7days, then randomly divided into7groups:normal control group,4℃7days group,4℃14days group,0℃7days group,0℃14days group,0℃7days herbs group,4℃7days herbs group. 2.2HerbsGancao ganjiang decoction add mahuang, formulation:gancao30g, ganjiang30g, mahuang20g. Mixing the three kinds of herbs and decocting, filtering, concentrating, containing0.2g drug of each milliliter decoction, bottling and placed4℃sealed conservation.2.3Index detection1) General state of health in mice, such as demeanor, activity, color and luster of fur and dietary, were observed.2) Lung tissue structure were observed with hematoxylin eosin(HE) staining method.3) Account the ratio of lung W/D.4) AQP-1protein expression in lung were measured by Western blot.5) Serum IgG and respiratory tract IgG were detected by ELISA kit.6) ExPression of BAFF mRNA in lung were detected by realtime quantitative fluorescence PCR.7) NF-κB p65protein expression in lung were measured by Western blot.2.4Statistical analysisAll data were analyzed by SPSS16.0statistical soft ware. Experimental data were showed with mean±standard deviation. T test and ANOVA was used to compared the differences. P<0.05level was used to indicate statistically significant difference.3. ResultsThe mouse of4℃7days,4℃14days,0℃7days group and0℃14days group became pale, less active, and reduced weight gain, part of mouse’s weight and food intake gradually reduced. Lung pathological section revealed that the mouse lung tissue of4℃and0℃7days group became edema, congestion, alveolar space had plenty of water fluid retention, and visible infiltration of inflammatory cells. The results were consistent with the evidence of exogenous cold-evil injury lung, and lung lesions were obvious. The animal model of exogenous cold-evil injury lung was success.Weighing the mice wet and dry lung and account the W/D ratio. We found the W/D ratio of4℃and0℃7days group increased (P<0.05) and the expression of AQP-1descended compared with the normal control group, we speculated that the exogenous cold-evil can interfere lung water transport and the mechanism may be down-regulating expression of AQP-1protein.Compared with the normal group, IgG-S and IgG-R concentra-tions of4℃7days group,4℃14days group,0℃7days group and0℃14days group were significantly increased, the BAFF mRNA expression and NF-κB p65subunit expression also raised. Combination the mice lung histopathology results, we speculated that the exogenous cold-evil destroyed the mice lung defense function, damaged the lung, which triggered the body’s acquired immunity and made the secretion of antibodies increasing. Its mechanism may be through up-regulating expression of BAFF and activate NF-κB signaling pathway.The mouse pulmonary pathological of4℃and0℃for7days which herbs treated significantly recovered, and the expression of AQP-1increased.