Experimental Research Of Cold And Cold Drink Lung Injury Theory And Pathogenesic Mechanism

Objective“Cold lung injury” and “cold drink lung injury”is a classicthe etiology and pathogenesis of theory. Form cold and cold drinksare from the outside feeling cold evil, whether it is a unilateralpathogenic factors “cold water” and “cold”, or both of thepathogenic factors superposition “form cold cold drinks” are easyto hurt lung disease, as the main reason of disease of lead to cough.In recent years, the academia of neijing lung “cold cold drinks”are also have more reports of clinical disease, but about the coldand cold drink pathogenic experiment research,especially thecomparative research is less. Therefore, this topic proposed underthe guidance of TCM etiology and pathogenesis theory, simulate theeffect of factors such as temperature, time, put the micerespectively in a cold environment, feeding the cold water, in thepreparation of “cold”,“cold drink” animal model, and using thelung as the research target organs, snapping the research datacomparison, observation form cold and cold drinks and form cold colddrink in mice lung main air breathing, main line function such aswater, toward the lotaustralin and who, and by measuring therespiratory mucopolysaccharides, mucosal immune molecules IgG, SIgA, TNF alpha, sticky protein MUC5AC and lung AQP-4and indexesof whole blood viscosity, plasma viscosity, with “cold” comparedwith “cold water” as the breakthrough point, to explore neijing“lung”cold cold drinks are the theoretical connotation andmechanism, and provide experimental basis for guiding clinicalapplication. This topic has theoretical innovation and clinicalguidance value.Methods1. Theory researchIs winter cold gas, is one of the six qi, normal climate changeis nature, if climate change anomalies, cold last time is too longon the human body, hurt and cause disease, cold at this time it iscalled a cold evil.Cold evil pathogenic also seen in other seasons,such as temperature, wading in the rain, sweat out when the wind,such as air conditioning too cold or too cold, summer is often animportant factor for feeling cold evil.Cold evil pathogenic differin typhoid fever, cold,"typhoid fever" the cold evil and muscleinjury, Outside the sun be the spirit of suppressed sun be the spiritof evil in the outer table of the cold syndrome, known as "cold"of the pathogenic factors."Cold" in the cold evil straight in,injury and sun be the spirit of the viscera, block viscera qiactivity, and blood running in the performance of cold syndrome,is a “cold drink” factors caused by disease.Lung being the delicate organ, its main physiological functionis the main air breathing, the main lines of water, lotaustralin,section chief, dispersing pertains transforming physiologicalcharacteristics.By dispersing pertains and transforming thecoordination function, make the airway unobstructed, breathing to make, to the normal exchange of gases in vivo and in vitro;Of thebody lose, run, and excretion of water and the dredging andregulating role;By dispersing pertains to who gas on body surface,make the gas can be adjusted to control the open closed, coujustified who make sweat discharge in moderation, can realize thefunction of guard outside the royal evil.The lung main gas,breathing, begin to understand in the nose, and outer coat, canopyfor five zang-organs, not to cold heat, is a charming is dirty.“Cold” and “cold drinks are cold evil”, the most easily hurtthe sun be the spirit, so the body feeling cold evil often firstmade the lungs, the way such as cold evil by fur, nose and mouthorgan invasion of the lung, easy to damage the lungs.In the six evilsof the cold evil lung injury in the most rare.The difficult, the49, difficult, also recorded“form of cold drink cold injury lung”,points out the exogenous cold evil, cold cold drinks can hurt thelungs; Canon said,“the cold eat into the stomach, as to pulmonarylung cold” from the pulmonary vein;“Heavy cold injury lung”.“Cold" that is cold and cold drinks.Cold and cold drinks are cold evil, easy injury sun be the spirit,because of different pathogenic factors of lung physiologicalfunctional disturbance, resulting in different clinicalmanifestations.Lung cold is the main way a associated with fur,because of lung in vivo for hair, fur cold exogenous cold injurylung performance for table empirical;The other one is associatedwith lung and stomach meridians, For cold drinks from the nose andmouth into the stomach, from pulmonary vein into the lungs, canappear lung and stomach’s sun be the spirit virtual failure andclinical symptoms of Yin qi KangCheng.Form cold lung injury cause lung qi can’t spread upward, lung can’t promote the water running,surface of the sun be the spirit is suppressed, nasal congestionand afraid of cold and no sweat pores. Fluid to gather to drink,breathing disorders and the bosom is frowsty asthma cough;Treatmentwith divergent wind chill, give priority to promote lung. Cold drinklung injury to the divergence of lung qi not, main show is Yin qiKangCheng and the conditions of the sun be the spirit is weak.Sunbe the spirit of the lung is weak, lack of lung qi is the functionof outward and inward dereliction of duty, gasification functiondisorders cause lung qi cannot push fluid running and form phlegmretention, phlegm retention blocking the airway, affect the gasexchange, cough, wheezing, even cannot lay down, drink treatmenttemperature on the lungs. Clinically for the treatment of bronchialasthma, chronic obstructive emphysema and pulmonary interstitialdiseases, chronic cough such as cold drink patients with lung injury,can use this method to treat.If cold drinks and exogenous cold evil(namely cold cold drinks) and single factor form cold or cold drinklung injury degree from lung cold, cold cold drink lung injurydegree deeper, Lung qi can’t spread upward and outward, sun be thespirit damage of lung and stomach, lung governance and regulationof physiological function and the cases of gas production andoperation is affected, cases of gas production, qi activity,promote blood flow, blood running, appear XiongGe full tightness,breathing, blood and fluid stay, such as treatment on releasinglungs, cold temperature lung drinks to give priority to.2Experiment study2.1Animals and grouping40specific pathogen free Kunming mice,17士lg, male and female in half,Conventional breeding ten days were randomly divided into4groups:(1) under normal temperature control group,(2) cold group,and (3) cold drinks group,(4) shape cold cold drinks group. Ingroups of more than15days later after anesthesia, broken neckdeath animals specimens-80℃freezer for later use.2.2Index detection1）General observationObserve the mice bearing, diet, activity, lip color, fur andweigh, anal temperature. In the case of a change2） Structure of lung tissue in mice to observeUsing haematin eosin (HE) staining, light microscopeobservation.3） AQP-4lung tissue expression of determinationUsing Western blot method.4） Airway glycosaminoglycan detectionCarbazole colorimetric method5） Bronchoalveolar lavage fluid of TNF alpha and airwayliquid IgG SIgA detectionBy ELISA kits6） Airway mucins MUC5AC detectionBy real-time fluorescent quantitative PCR detection7）Blood rheology detectionUsing heparin anticoagulant tube routine eye is about1.2ml ofblood in mice, the instrument automatically detect whole bloodviscosity (high shear viscosity and low shear viscosity) and plasmaviscosity, repeat each sample test3times, the instrument recordas a result, the averaging. 2.3Statistical analysisAll data were analyzed by SPSS16.0statistical soft ware.Experimental data were showed with mean士standard deviation. Ttest and ANOVA was used to compared the differences. P<0.05levelwas used to indicate statistically significant difference.Results(1)General condition:Normal control group on day1to15daysmice lip ruddy, hair luster, ears and tail are light pink, appetiteis normal, weight increase gradually, smooth breathing, responsive,anal temperature is normal, normal quality;Shape of cold, colddrinks, form cold cold drinks group mice appeared lips, ears, tailby gradually become pale pink, by the first fast breathing, slowdown gradually, after diet is reducing gradually, defecate textureis soft and wet.Anal temperature decreased gradually, graduallylosing weight.Shape on day1to7days in cold mice body weightincrease gradually, respiration, activities are very active,vertical reaction, MAO extrados curled, lumpy, chills, oral oweruddy, owe hair luster;Cold drinks group also showed depression,unresponsive, diet is reducing gradually, defecate pond.Shape ofmice cold cold drink lips, ears and tail owe ruddy, weight shallbe gradually increased, after gradually reduce, respiration, analtemperature and temperature are gradually decline, stool soft.Onday8to15days, normal temperature, diet, activities, mainland,though the control breath, body weight and anal temperature arenormal.Cold mice appeared spiritual malaise, curled up less still,breathing slow, diet is reducing gradually, lips, ears and tailgradually become pale, though weight lifting, anal temperaturegradually returned to normal.Cold drinks, cold cold drinks groupmice started to gather together, tongue color clear is dark purple, ears and tail are dark red, though slow to respond;Breathing is alsogradually slow down, gradually reduce the amount of diet, analtemperature and decrease its weight, shape breathing more weak,15th day cold cold drinks group "inactivity, even refusing foods,angular, fur lackluster, lips, ears of pale in color.Analtemperature and weight compared with the form of a cold, cold drinksgroup numerical minimum, defecate texture soft, wet rotten symptomssuch as a heavy7days.(2) Weight:In building7days to15days, normal control groupmice weight over time weight value showed a trend of increasegradually.Cold, cold drinks group and form cold cold drinks groupweight over time gradually reduce weight value.Compared with normalcontrol group, cold, cold drinks group and form within7days ofcold cold drinks group mice weight is on the high side, thecomparison between groups weight, increased weight the most obviousform cold group, cold drink set times, shape of cold cold drinksgroup again;Shape of cold, cold drinks group within15days ofweight were higher than in normal control group, the shape is coldcold drinks group mice weight value is lower than the normal controlgroup, group comparison between weight and shape cold cold drinksgroup, weight loss, the most obvious form of cold group weighthigher than cold drinks group.(3) Anal temperature:As building the extension of time, thenormal control group, cold, cold drinks, form cold cold drinks group,all the mice in the seventh day of the anus temperature less than15days.In building7days, compared with normal control group, cold,cold drinks group, form cold cold drinks group were lower,comparison between groups, the shape of mice cold cold drink anal temperature decrease, the most obvious cold drinks group times,shape of cold group again;In building15days, compared with normalcontrol group, the shape of mice cold anal temperature on the highside, cold drinks and cold cold drinks group reduced, the comparisonbetween groups, reduce the most obvious form cold cold drinks groupmice, cold drink set times, shape of cold group again.(4)Pathological results:Then pathological lung tissue showednormal control group, cold drinks group, cold, cold cold drinksgroup of pulmonary lesions degree according to the order ofincreasing trend, to form cold cold drinks group is mostobvious.Shape of cold times, cold drinks group again.:(1) undernormal temperature control group, bronchial lung tissue, thealveolar structure is clear, bronchial epithelial integrity,medium and small neat bronchial mucosa epithelial cells, pulmonaryinterstitial inflammatory cells infiltration, hyperplasia ofalveolar walls are not seen, alveolar interval no hyperemia andedema, alveolar space to expand, and there is no liquidleakage.Layer (2) the form of a cold groups: bronchial mucosa andsubmucosa have mild to moderate inflammatory cells infiltration,outer membrane layer is part of the inflammatory cells infiltration,interstitial have a large number of inflammatory cells infiltration,congestion and edema.(3) cold drinks group: bronchial submucosa andouter membrane layer are meet mild to moderate inflammatory cellsinfiltration, with mild interstitial inflammatory cellinfiltration, and part of the lung tissue necrosis.(4) shape coldcold drinks group: bronchial submucosa of inflammatory cellsinfiltration, lymph nodes of lung interval with lymphocytesconstitute sample structure, it is also seen as part of neutrophils constitute sample abscess structure.(5) Test results of lung tissue AQP-4expression:Western blotmethod to detect cold form groups, cold drinks and cold cold drinksgroup of AQP4protein expression of lung tissue in mice weresignificantly lower than normal control group;Compared with colddrinks group, cold type group and form cold cold drinks group ofAQP4protein expression decreased lung tissue, in order to form coldcold drinks group decreased significantly, fractal cold group takesecond place;Compared with group form cold, cold drinks group ofAQP4protein expression of lung tissue on the high side, form coldcold drinks group of AQP4protein expression to reduce the lungtissue.Tip shape cold and cold drinks may cut AQP4expressionthrough affect the lung function of the main communication channel,thereby aggravating alveoli and interstitial edema, cold and colddrink to form two kinds of pathogenic factors in the role of themost obvious, cold times, cold drink again.(6) Test results of airway mucopolysaccharides:Make mould withcarbazole colorimetry to detect each group of mice airway D-glucose aldehyde lactone numerical changes, according to the airwayof mice pathological and histological results show that the shapeof cold bronchial mucosa, submucosa and see more inflammatory cellsinfiltration in the outer membrane, cold drinks group accidentalbronchial mucosa inflammatory cell infiltration, form cold colddrinks group can see bronchial submucosa see inflammatory cellinfiltration in the part, combining the experiment airway D-glucose aldehyde lactone numerical results show that the shape ofcold, cold drinks and cold cold drinks group compared with normalcontrol group, are rising.Compared with form cold group, cold cold drinks group ratio on the high side, low cold drinks group.Comparedwith cold drinks group, form cold cold drinks and cold group ratioalso increased, in order to form cold cold drinks group of the mostobvious, the results confirmed that the form of a cold cold drinkcan indeed make airway secretions increase, lead to airway tissuelesions in mice, the degree of pathological changes in cold colddrinks in the most obvious, cold times, cold drink again.(7) Respiratory tract mucosal immune molecules of testresults:Mice by ELISA kits each numerical airway in bronchoalveolarlavage fluid IgG, results show that compared with normal controlgroup, cold, cold drinks group and form airway express value liquidIgG cold cold drinks group were significantly increased.Comparedwith cold drinks group, form cold cold and cold drinks groupexpressed higher values, to form cold cold drinks group expressionvalues increased most obviously, cold times;Compared with form coldgroup, form cold cold drinks group ratio increases, cold drinksgroup expression value is low, tip shape cold and cold drink canmake produce lots of IgG in serum and lung less susceptible toinvaded assault, consistent with TCM lung function outside the Lordwho, in cold and cold drinks in both the role of the more obvious,cold times, cold drink again.Airway liquid results showed that content of TNF alpha:Airwayliquid content of TNF alpha results showed that compared with normalcontrol group, each module of the concentration of TNF alpha weresignificantly increased;Compared with cold drinks group, form coldcold and cold drinks group all rise, rise to form cold cold drinksgroup; the most obviousCompared with form cold group, form cold colddrinks group ratio increases, low ratio of cold drinks group, tip shape cold and cold drink can induce lung infection, the mostobvious, the extent to form cold cold drink cold times, cold drinkagain.Airway of SIgA concentration results showed that compared withnormal control group, cold group, cold drinks and form cold colddrink SIgA expression values were significantly lower in group ofairway liquid;Compared with cold drinks group, form the expressionof cold cold and cold drinks group values are lower, in order toform cold cold drink the most obvious;Compared with group form cold,cold drinks group expression values rise, form cold cold drinksgroup expression values fell, the tip shape of cold and cold drinkcan reduce respiratory tract mucosal immune molecules SIgAexpression, cause respiratory tract infections, and even developinto pneumonia and systemic infection, cold and cold drink to formin the pathogenic factors of lung injury is more serious.From airwaymucosa secretion level comparison, cold drinks can cause airwayliquid produced a large number of SIgA, lesions in the trachea, alsocan cause the lung damage,"water main line lung function isaffected, the fluid generated and metabolic dysfunction and stopthe formation of phlegm retention.(8)Airway mucins MUC5AC test results:By real-time fluorescentquantitative PCR testing groups of mice were respiratory tractMUC5AC numerical changes, the results show that compared withnormal control group, cold, cold drinks group, form airway mucinprotein expression of cold cold drinks group were significantlyincreased;Compared with cold drinks group, form cold cold and colddrinks group expression values were elevated, to form cold colddrinks were significantly increased;Compared with form cold group, form cold cold drinks group of airway mucus proteins expressionvalues on the high side, cold drink below form cold group, tip shapecold and cold drink can make sticky protein MUC5AC highly expressedin the airway and lung tissue, cause many severe respiratory diseaseand lung function damage, the degree of damage to form of cold andcold drinks in the role, the most obvious form cold times, cold drinkagain.(9) The test results of blood rheology:Hemorheology testexperiment results show that, compared with normal control group,each group whole blood viscosity are high;Compared with form coldgroup, form cold cold drinks group of whole blood low shearviscosity (10s-1) on the high side, to form cold cold drinks groupwhole blood low shear viscosity increased most obviously;Comparedwith cold drinks group, cold type group and the form of a cold colddrink the whole blood low shear viscosity (10s-1) are on the highside, in order to form cold cold drink of whole low shear viscosityon the high side, the most obvious to test to measure the aggregationof red blood cells, choice under the condition of the low shear ratedetecting the change of hemorheology between groups numerical,results suggest form cold and cold drink can make blood viscosityincreases, further to make the body microcirculation and thrombosis,cause blood stasis, from the perspective of Chinese medicine todescribe influence the function of the lung in lotaustralin, accordwith haemorrheological nature pathogenic characteristics of bloodstasis, its damage degree to form cold cold drinks in the mostobvious, cold times, cold drink again.ConclusionComprehensive the above results, it can be found form cold and cold drinks in stimulation treatment in mice after7days, eachmodule of anal temperature values are lower, weight is on the highside, gradually with the extension of cold evil duration, eachbuilding module mice are high in dealing with15days after the anustemperature, weight lifting, mouth, ears, skin color in mice by pinkto pale;Mice huddled together, lumpy, chills,"inactivity,breathing by quickly become weak gradually, weight lifting, theabove show the symptoms of cold and cold drinks are a sign of a seriesof cold, can cause the body performance its degree of pathogenicfactors to form cold and cold drink both overlap with most seriousimpact on the body, cold drinks, form the cold again.According toeach building modules of lung tissue pathological results suggestform cold and cold drinks all can hurt the lungs, and the degreeof pathological changes in lung to form cold cold drink both themost severe pathogenic factors superposition, cold drinks, form ofcold again, various made module mode is a success.Mucopolysaccharides through respiratory tract, mucosal immunemolecules can be found that a single factor of "cold","cold" andboth of the pathogenic factors superposition "shaped cold colddrink" invaded can damage the lungs, all can be induced by pulmonaryinfection of pathogenic factors, can seriously affect d-glucosealdehyde lactone (MS), and a variety of inflammatory cytokines IgGSIgA and the expression of TNF alpha, reduce the body’s immuneprevention effect, lead to heavier lung infection and systemicinfection, the degree of lung infection in cold cold drink, theworst form of cold, cold drink again;Can be found by observing thelung tissue AQP4protein expression form cold and cold drink canseriously affect the airway and lung tissue of AQP4expression, increase airway inflammation, affect the lung function of thewaterway, and further affect the lung respiratory function, thedegree of pathological changes in cold cold drink, the most obviousform cold times, cold drink again.For each different in bloodrheology detection module, based on the results form in cold colddrink both can make the mice increased blood viscosity, the wholebody blood flow resistance increases, easy to form themicrocirculation and blood clots, increase decreased respiratoryfunction and low oxygen, and the result accords with the nature ofthe "cold sex grinds" and pathogenic characteristics, itspathogenic degree to form cold cold drink two kinds of pathogenicfactors superposition, the most obvious form of cold times, colddrinks group again.In a word, form cold and cold drink etc invaded all can causedamage to the lungs and the degree of disease.Form cold and colddrink etc invaded all can promote the expression of a variety ofinflammatory cytokines and affecting airway mucopolysaccharidessecretion disorder, make the body’s immune defense function decline,lead to vulnerable invaded by diseases and disease, degree ofillness to form cold and cold drink both overlap with the mostserious pathogenic factors, cold times, cold drink again.