Experimental Study On The Inhibition Of Mapk And NF-κB Signaling Pathway Of The Polysaccharide Based On Airway Inflammation Of Asthma

Purpose:Through the OVA induced bronchial asthma mice model is set up, And give her ABPS treatment, study ABPS on the effect of bronchial asthma airway inflammation and its possible mechanism.Methods:32 normal female BALB/c mice, only 18+/-5 g weight, were randomly divided into 5 groups, each group of eight, the normal control group, asthma model group, lower dose treatment groups, ABPS, ABPS high dose treatment group, Dexamethasone, positive control group. Except the normal control group, the other 3 groups in first days, seventh days and fourteenth days in 3 intraperitoneal injection of 1ml antigen liquid (containing egg protein 100mg, aluminum hydroxide powder sensitized 100mg), normal control group sensitized with physiological saline instead of. The twenty-second day the mice were placed in the incompletely closed box, with 1% ovalbumin (OVA) solution with the atomizer inhalation,1 times a day, every 20 minutes, for 4 consecutive days. The normal control group with physiological saline instead of ova. Every 1 hours before the given aerosol inhalation of Agaricus blazei polysaccharide intervention, polysaccharide low dose group received 200 mg/Kg orally, polysaccharide high dose group was given intragastric administration of 400mg/Kg, the normal saline group and asthma model group were fed with the same amount,1 times a day, a total of 10 days. Twenty-fifth days,24 hours after the last challenge, the mice were with ether anesthesia, the specimens according to the requirements. The last sensitization to strengthen executed in mice after 24 h ether anesthesia, cut the neck skin, and cut a small opening in the center of the trachea, inserted into the trachea cannula. Line with phosphate buffer 4 ml bronchoalveolar lavage, wash 3 times back and forth, collecting bronchoalveolar avage fluid (BALF) and lung tissue samples, etc. By enzyme-linked immunosorbent (ELISA) and western blot detection of bronchoalveolar lavage fluid (BALF) of IL-4、IL-5、IL-13 levels, and observe the inflammatory cells in BALF and lung tissue pathology change. By using ELISA determination of cytokines in BALF, IL-4、IL-5、IL-13 by HE staining and PAS staining to observe each group of mice lung pathological changes such as tissue inflammatory cell infiltration. Immunohistochemical staining to observe the NF-kappa B p65 proteins in the lung tissue expression.Using western blot method to detect each group in the lung tissue of mice p-ERK, p-JNK, the expression of p-p38MAPK conditions.Result:(1) compared with normal control group, the total number of inflammatory cells in BALF of asthma model group mice and PMN, Eosnophils, lymphocyte count is increased significantly, the significant difference (P<0.05); ABPS is low, the high dose group mice inflammatory cells in BALF and PMN, Eosnophils, total lymphocyte count compared with asthma model group significantly decreased, the significant difference (P<0.05), compared with normal group, BALF of asthma model group mice IL-4、IL-5、IL-13 levels, significant difference (P<0.05); ABPS is low, the high dose group mice cytokines IL-4、IL-5、IL-13 in the BALF, IL-4、IL-5、IL-13 levels, compared with asthma model group, significant difference (P<0.05) and (2) by observing the pathological lung tissue HE and PAS staining dye, asthma model group mice bronchospasm, luminal stenosis, epithelial cell swelling, fall off, bronchi and vessels surrounded by a large number of inflammatory infiltration, give priority to with EOS, lymphocytes and PMN, edema of mucosa and submucosa. ABPS is low, the high dose group mice lung tissue pathology change of asthma model group significantly reduce. No obvious lung tissue pathological changes in normal mice. ABPS comparison between low and high dose group, no significant difference (P> 0.05). (3) by observing the immunohistochemical staining:the NF-kappa Bp65 positive expression level significantly increased in asthma model group; And in her positive ABPS extracts of two groups of protein expression level decreased obviously. (4) western blot test results showed that the expression of p-ERK and p-p38MAPK in asthma model group is significantly higher than the control group (P<0.05); In her ABPS expression of two group was obviously lower asthma model group (P<0.05); In model group p-JNK expression was significantly higher, but no obvious difference compared with drug group.Conclusion:ABPS could inhibit asthma in the lung tissue of mice p-p38MAPK, p-ERK, p-JNK and NF-kappa Bp65 protein expression, downgrade IL-4、IL-5、IL-13 level, effectively restrain the airways of the asthma mice inflammatory cell infiltration, so as to reduce airway inflammation.