Effect Of Lijinfang On HDAC Protein And Its Downstream IL-6 And TNF-α In COPD Rats With Fei-qi Deficiency Syndrome

Objective:The aim of this study was to prepare（COPD）disease rats with deficiency of Fei-qi deficiency syndrome（FQDS）by smoking combined with endotracheal injection of lipopolysaccharide（LPS）.Using Western blotting（WB）and enzyme linked immunosorbent assay（ELISA）to observe the effect of HDAC-2 content in lung tissue and serum and IL-6 in the lower reaches of alveolar lavage fluid in rats with FQDS COPD.To study the changes of hdac-2,il-6 and TNF-αin Lijinfang treated rats.To explain the effect of Lijinfang from the aspect of anti-inflammatory mechanism,and to provide a real and objective scientific experimental basis for the treatment of COPD with Chinese medicine.Methods:1.Preparation of a model of COPD with FQDSSixty-six male 7-week-old SPF Wistar rats were randomly divided into 6groups:group A（normal group,n=11）,B group)（model group,n=11）,C group),and group A（n=10,n=11,n=11）,and group A（control group,n=11,n=11）.Group E（Lijin Fang high dose group,n=11）,F)（erythromycin（EM）group,n=11）.The 6 groups were fed adaptively for 7 days,fed freely and drinking water.7 days later,the normal group was fed according to the routine,and no intervention.B,C,D,E,etc.In group F,lipopolysaccharide（LPS）was injected into the trachea according to the smoking method and LPS,was injected into the trachea on day 1 and day 14,respectively,at the same time,the model was made on the 2nd and 13th day.A rat model of FQDS（COPD）was established by cigarette smoking in a self-made smoke box for 28 days.2.Erythromycin,Lijinfang interventionGroup A,group B was given saline 5ml/kg^-1d^-1),group C was given low dose 4.48g/kg^-1d^-1,group D was given medium dose 11.2g/kg^-1d^-1.High dose of 22.4g/kg^-1d^-)was given to group E and EM 30.8mg/kg^-1d^-1was given to group F for 30 days.3.Specimen collection and detectionAfter anesthesia,blood samples were taken from abdominal aorta,bronchoalveolar lavage fluid（BALF）was collected from right lung and lung tissue was collected from left lung.After the collection of samples,the relative expression of HDAC-2 in left lung tissue was detected by Western Blot（WB）method,the pathological changes of rat lung tissue were observed by optical microscope,and the serum IL-6,was measured by enzyme linked immunosorbent assay（ELISA）after right lung BALF and blood centrifugation.TNF-αcontent.Results:Rat model preparation:According to the general emotion of hair,urination,defecation and active state in rats,the model of FQDS in COPD was accorded with the model of FQDS in rats.The left lung tissue was observed by light microscope.All the pathological changes of the left lung were in accordance with the pathological changes of COPD except group A,and the pathological changes of the drug intervention group were lighter than those of group.The results of pathological score of lung tissue inflammation were as follows:（1）The results were as follows:（1）compared with group A,the inflammatory pathological reaction of lung tissue in COPD model group（group B）was significantly higher than that in group A（P<0.05）.（2）compared with group B,the inflammatory pathological reaction of lung tissue in group C,D,E,F was significantly lower than that in group B（P<0.05）.（3）the improvement degree of lung inflammation in group E was better than that in group C,D and E（P<0.05）,and there was a significant difference among the three groups（P<0.05）.There was no significant difference in inflammatory pathological scores between group C and group D（P>0.05）.（4）there was no significant difference in inflammatory pathological scores between group E and group F（P>0.05）,and there was significant difference between group E and group F（P<0.05）,and there was significant difference between group E and group F（P<0.05）,and there was no significant difference in pathological scores of lung tissue between group E and group F（P<0.05）.High dose of Lijinfang and EM could not recover the pathological changes of lung tissue to normal level.Results of drug interventions:（1）The results were as follows:（1）compared with group A,the expression of HDAC-2 in lung tissue of COPD model group（group B）was lower than that of control group（group A）,and the expression of IL-6,TNF-αin serum and BALF of rats was significantly higher than that of group A（P<0.05）.（2）compared with group B,group C,D,E,group F,three groups of Lijinfang（C,D,E）and EM group（F）could upregulate the relative expression of HDAC-2and decrease the expression of IL-6,TNF-αin lung tissue.And low,middle,high-dose Lijinfang intervention group（C,D,E group）in pairwise comparison,with the increase of dosage,the more obvious effect on up-regulating the expression of HDAC-2 in lung tissue and inhibiting the IL-6,TNF-αof serum and BALF.The difference was statistically significant（P<0.05）.（3）compared with F group,there was no significant difference between E group and EM group（P>0.05）.（4）compared with group A,the IL-6,TNF-αof HDAC-2,serum and BALF in group E,F was significantly different from that in group A（P<0.05）.High dose of Lijinfang and EM could not completely inhibit IL-6,in lung tissue of rats（P<0 05）.TNF-αand HDAC-2 returned to normal level.Conclusions:1.Smoking combined with endotracheal injection of lipopolysaccharide（LPS）was used to make the rat model of FQDS（COPD）successfully.2.Lijinfang can increase the expression of HDAC-2 in lung tissue of COPD rats with FQDS,decrease the release of IL-6,TNF-αinflammatory factor,alleviate the inflammation of airway and lung tissue,improve the pathological changes of lung tissue in COPD rats,and delay the process of COPD.