The Effect Of Kechuanning On Expression Of Chemokine-like Factor1(CKLF1) In Asthmatic Rat

Objective:To establish a Immunohistochemisty method and a reserve transcription-polymerize chain reaction(RT-PCR) method for detecting of CKLF1expression level in the lungs of asthmatic rats.To investigate the effect of Kechuanning on the CKLF1expressions well as to analyze its role in asthma Pathogenesis.Methods:50SD rats labeled with weight in random categorization were divided into a normal group(group A) and a modeling group;the modeling group were sensitized by ovalbumin (OVA) and Al(OH)3,and the rats were made by breathing in5%OVA continuously for2weeks.As rats models were established.The rats were subdivided into three group.B was asthma model group.C was Budesonide group.D was Kechuanning group.The rats in group B were treated with inhaled Physiological Saline once a day for7days;the rats in group C were treated with inhaled Budesonide,once a day for7days;and the rats in group D were treated with kechuanging liquid for inhalation, once a day for7days.During the process of experiment,we observed the ordinary situation of the rats.And then the rats were executed within24h after final treatment. The total number of white blood cells and celluar distribution in bronchoalveolar lavage fluid,the pathology of lung were examined.The lung were used to test the expression of CKLF1mRNA via RT-PCR,and the CKLF1protein in lung were tested via immunohistochemisty.Rsults:1.The manifestation of rats after challenge:the rats were expressed as irritability,restlessness,scratching fur constantly,breathing rapidly and deeply,wheezing,cyanosis,controlled abdomen;reducing movement and responding bluntly.The rats lost the luster of hair and responded bluntly after sequent challenge.2. Analyze the cell numember of the bronchoalveolar lavage fluid (BALF):the total number of leukocytes, neutrophils,lymphocytes,and EOS of BALF were increased in group B(P<0.05);group D compared with group B,the total number of leukocytes,lymphocytes,and EOS were decreased(P<0.05);there were not significant differences between group C and D(P>0.05).3.The pathological changes in the lung tissue of rats:group A had no significant change;group B had significant inflammatory changes.Group C and group D were showed a mild inflammatory changes.4.The immunohistochemical result of CKLF1in the tissues of rat lungs in each group:in group A the CKLF1expressed weakly positive. In group B expression of CKLF1was significantly increased compared with group A.compared with group A,the expression of CKLF1was increased in Group C and group D,and decreased when compared with group B.There were no significant difference in group C and group D.5.The result of CKLF1mRNA in the tissues of rat lungs in each group: CKLF1mRNA expression level in group B was significantly increased compared with group A(P<0.05).Group C and group D had no significant difference(P>0.05),and decreased compared with group A(P<0.05).Conclusion:Kechuanning could reduce the inflammatory response of the lung tissue; in the lung tissue of asthmatic group CKLF1expression levels were significantly increased, Kechuanning could reduce the expression of CKLF1.