The Effect Of The Tonifying Vital Energy And Promoting Blood Flow Recipe On Pulmonary Apoptosis And Related Genic Proteinnum Bcl-2,Fas In Chronic Bronchitis Rats With Insufficiency Of Lung-Qi

Objective:To observe the effect of the tonifying vital energy and promoting blood flow recipe on pulmonary apoptosis and related genic proteinum of the rats,insufficiency of lung-qi of chronic bronchitis(CB),approach its functional mechanism,and explore the possible mechanisms,which will provide practical and theoretical evidence for developing new method of treating and preventing the insufficiency of lung-qi of CB.Methods:Take 50 healthy SD rats(paroecious about half)to raise in the experimental environment about one week.Adopt the improved smoking-fumigated to duplicate the model of the insufficiency of lung-qi of CB.The normal rats raise in the common environment without smoking-fumigated.After the model duplicate to succeed , divided into model group ,high-dose group of the tonifying vital energy and promoting blood flow,low-dose group of the tonifying vital energy and promoting blood flow, tonifying lung recipe group,and normal group without enay treatment(10 per-group and paroecious about half). Give the isotonic Na chloride or drugs to the five groups retain twenty-one days after the model duplicate,one time everyday.Normal and model group all drench the isotonic Na chloride,volume-10ml/kg;The volume of the high-dose group is 25.6g/kg(amount to the 14 multiples of the adult),0.18g/ml.The volume of the low-dose group is 12.8g/kg(amount to the 7 multiples of the adult),0.09g/ml.The volume of the tonifying lung recipe group is 8.63g/kg(amount to the 7 multiples of the adult),0.09g/ml.The rats in each group are put to be anaesthesia to death after two days of drench.Open the breast and take the lung tissue(0.5cm×0.5cm×0.5cm),which were fix with 4% formal -dehyde,embedded with paraffin,maken sections by HE and immunohistochemical method stain.Meanwhile take some lung tissues by FCM.Results:1Observation of the gemeral condition12 days after smoking-fumigated the rats in model group showed the symptoms,such as dyspnea,cough,wheezy phlegm losing brightness of hair,lassitud and so on.While the rats in normal group were without any symptoms.The weight in model group increased much slowly than normal group,at the last day of smoking-fumigated,weight addition in model group was(94.48±14.16)g,the normal group was(104.69±23.65)g,there were significant difference between the two groups(P<0.05). 2Observation of the lung's histomorphology2.1Macroscopic observation:The lung's appearance of normal group was common with color pink,surface smooth,elasticity good and lobes of lung edge sharpness.The lung's appearance of model group was abnormal with color gray-black,surface concavity,without burnish and ecchymosis were seen.2.2Obeservation under light microscope:The result of phthomorphology manifested that in normal group mucous membrane of bronchiole epithelia was integrity,cells lined up orderly,bronchiole dissepiment and alveoli had no distinctive pathomorphologic alteration under light microscope.In model group,bronchioles cavity was enlarged,vessel wall was thicked significantly,part of epithelial cellular necrosis and abscission,there was a lot of chronic inflammatory cells infiltration and venule with stagnant blood.3Bronchi and lung tissues'apoptosis of every group rats3.1Inflammatory cells'apoptosis of bronchi and lung tissues of every group rats by TUNEL and immunohistochemical method3.1.1Inflammatory cells'apoptosis of bronchi and lung tissues by TUNELCompare with normal group,the percentage of inflammato -ry cells'apoptosis of model group was decreased obviously,has statistics significance(P<0.05).Compare with model group,the percentage of every group to treatment was increased obviously,has statistics significance(P<0.01),the percentage of high-dose group compare with low-dose group was increased (P<0.01);the percentage of high-dose group compare with tonifying lung recipe group was increased(P<0.01);the percentage of low-dose group compare with tonifying lung recipe group wasn't have statistics significance(P>0.05).3.1.2Detect apoptosis gene protein bcl-2 with immunohistoche -mistyThe average grey level of the model group is obviously lower than the normal group , has statistics significance(P<0.01).Compaired with model group,the average grey level of each treatment group is obviously elevates,it has statistics significance between each group(P<0.01),the average grey level of the high-dose group is higher than the low-dose group,has statistics significance(P<0.01);Between the high-dose group and tonifying lung recipe group,the average grey level is elevates, it has statistics significance(P<0.01);Between the low preparation group and tonifying lung recipe group,it has statistics significance(P<0.05).3.1.3Detect apoptosis gene protein fas with immunohistochemis -tyThe average grey level of the model group is obviously higer than the normal group , has statistics significance (P<0.01).Compaired with model group,the average grey level of each treatment group is obviously reduses,it has statistics significance between each group(P<0.01),the average grey level of the high-dose group is lower than the low-dose group,has statistics significance(P<0.01);Between the high-dose group and tonifying lung recipe group,the average grey level is reduses,it has statistics significance(P<0.01);Between the low-dose group and tonifying lung recipe group,the average grey level is reduses,it has statistics significance(P<0.05).3.2Detect each group of the rat's lung parenchyma with flow cytometry3.2.1Detect the rate of apoptosis through each group of the rat's lung parenchyma with flow cytometryThe model group is obviously higher than the normal group,has statistics significance(P<0.01).Compaired with model group,the rate of apoptosis of each treatment group is obviously reduses,it has statistics significance between each group(P<0.01),the rate of apoptosis of the high-dose group is lower than the low-dose group, but does not have statistics significance(P>0.05);Between the high-dose group and tonifying lung recipe group,the a rate of apoptosis is reduses, it has statistics significance(P<0.01);Between the low-dose group and tonifying lung recipe group,the rate of apoptosis is reduses ,it has statistics significance(P<0.05).3.2.2Detect cell cycle of each group of the rat's lung parenchym -a with flow cytometryG₀/G₁ period(silent period of cell/presynthetic phase):The cell percentage of the model group is obviously higher than the nomal group,has statistics significance (P<0.01).Com -paired with model group,the cell percentage of each treatment group is obviously reduses,it has statistics significance between each group(P<0.01),the cell percentage of the high-dose group is lower than the low-dose group,but does not have statistics significance(P>0.05);Between the high-dose group and tonifying lung recipe group,the a rate of apoptosis is reduses, it has statistics significance(P<0.05);Between the low-dose group and tonifying lung recipe group,the rate of apoptosis is reduses,but does not have statistics significance(P>0.05).S period(synthesis period of cell):The cell percentage of the model group is obviously lower than the nomal group,has statistics significance(P<0.01).Compaired with model group,the cell percentage of each treatment group is obviously elevates,it has statistics significance between each group(P<0.01),the cell percentage of the high-dose group is higher than the low-dose group, but does not have statistics significance(P>0.05);Between the high-dose group and tonifying lung recipe group , the a rate of apoptosis is obviously elevates,it has statistics significance(P<0.05);Between the low-dose group and tonifying lung recipe group,the rate of apoptosis is elevates , but does not have statistics significance(P>0.05).G₂/M period(post-synthetic phase of cell/mitotic period):The cell percentage of the model group is obviously lower than the nomal group, has statistics significance(P<0.05).The cell percentage of the high-dose group is obviously higher than the model group,has statistics significance(P<0.01);Between the low-dose group and the model group,the rate of apoptosis is elevates , but does not have statistics significance(P>0.05);Between the tonifying lung recipe group and the model group,the rate of apoptosis is obviously elevates, has statistics significanc(eP<0.05).Between each treatment group,there is no statistics significance(P>0.05).PI:The model group is obviously lower than the normal group,has statistics significance(P<0.01).Compared with the model group,eash treatment group is obviously elevates,there is statistics significance during the group comparison(P<0.01),the high-dose group is higher than the low-dose group, but does not have statistics significance(P>0.05);Between the high-dose group and tonifying lung recipe group,the a rate of apoptosis is obviously elevates,it has statistics significance(P<0.05);Between the low-dose group and tonifying lung recipe group,the rate of apoptosis is elevates , but does not have statistics significance(P>0.05).Conclusion:1 Under the condition around insufficiency of CB lung-qi,inflammatory cells'apoptosis of bronchi and lung tissues existed apoptosis to be delaved,The express was up-regulation that apoptosis related genic proteinum bcl-2 and down-regulation that genic proteinum fas . The bronchi and lung tissue'parenchyma cells existed excessus apoptosis that lead to the breakdown of bronchi and lung tissues'architectonic,that is one of the evolutionary mechanism about insufficiency of CB lung-qi.2 The tonifying vital energy and promoting blood flow recipe can increase percentage about the inflammatory cell'apoptosis of bronchi and lung tissues by abaissement the express of genic protein bcl-2 and up-regulation the express of fas, promote to discard the inflammatory cells to depend on apoptosis,which is very important to disappear the inflammatory reaction.3 The tonifying vital energy and promoting blood flow recipe can speed up the synthetical rate of lung parenchymary cells by hold-back its apoptosis , then can anastate the lung parenchymary numbers and function,strengthen its recovery capability by itself,which assign new connotation to cure insufficiency of lung-qi of CB.