| Purpose:To explore the effect of Traditional ChaoYi Medicine JingFangBaiDu Powder on the expression ofMAPK(mitogen-activated protein kinase)/NF-kB(nuclear factor-k-gene bindin)signaling pathway in asthmatic mice.Methods:We randomly divided sixty BALB/c male mice of clean grade into 5 groups,twelve in each group:normal control group,asthma model group,ChaoYi Medicine JingFangBaiDu Powder high dose group,ChaoYi Medicine JingFangBaiDu Powder low dose group,dexamethasone positive control group.After 7 days of adaptive feeding,the mice were intraperitoneally injected with a mixture of ovalbumin 300 ? g,alum 3 ml,and 0.9%sodium chloride 15 ml,200?l per injection.we sensitized the other 4 groups of mice on days first,seventh and fourteenth,besides the normal control group was injected with 0.9%sodium chloride injection at 200 ?l.On the 23rd day,the mice were given 5%OVA(egg white)inhalation for four consecutive days,but the normal group was inhaled with 0.9%sodium chloride injection under the same states.The mice in JingFangBaiDu Powder high dose and low dose group and dexamethasone positive control group were given intragastric administration for 10 consecutive days from the seventeenth day.The normal control group and the asthma model group were intragastrically administered with 0.9%sodium chloride injection.Twenty-four hours after the last challenge,the mice were sacrificed and their lung tissue and BALF(Bronchoalveolar Lavage Fluid)were taken.Pathological changes of lung tissue were observed by HE and PAS staining.The inflammatory cell types and counts in BALF were observed by Diff-Quick staining,the levels of IL-4(Interleukin-4),IL-5(Interleukin-5),IL-13(Interleukin-13)and IFN-?(Interferon-?)were determined by ELISA.The effects of JingFangBaiDu Powder on the expression of p38MAPK and NF-?Bp65 in lung tissue of asthmatic mice were detected by immunohistochemical staining and western blotting.Results:1.We can observe from the lung tissue section of each group of mice by HE and PAS staining,compared with the normal control group,the airway wall and smooth muscle in the asthma model group were significantly thickened,the submucosa was widened,the mucosal epithelium was hyperplastic,the lumen is blocked by mucus,and goblet cell hyperplasia;the above-mentioned changes in the ChaoYi Medicine JingFangBaiDu Powder high-dose group and the dexamethasone group were dropped compared with the model group,and the extents of lung tissue injury were lighter.2.Inflammatory cell types and counts in BALF were observed and counted by Diff-Quick staining.From the staining we can see that a large number of EOS(Eosnophils)was aggregated in the asthma model group,the total number of cells and three cell counts,including EOS,NE(neutrophilic granulocyte),and LYM(lymphocyte),escalated apparently,contrast with normal control group,there were apparent differences(P?0.05).ChaoYi Medicine JingFangBaiDu Powder high dose group and dexamethasone positive control group compared with asthma model group,EOS,the total number of cells and three cell counts decreased apparently(P?0.05).3.The ELISA method was used to detect various cytokines in BALF of each group of mice.The results showed that the levels of IL-4,IL-5 and IL-13 in the asthma model group were significantly higher than those in the normal control group,while IFN-? was significantly increased.The level of the drug was decreased(P?0.05);the levels of IL-4,IL-5,and IL-13 in Chaoyang Jingfang Baidu Powder low,high dose group,and dexamethasone positive control were higher than those in the asthma group.Decreased while IFN-? levels increased significantly(P<0.05).4.Western blot analysis of p38MAPK showed that the level of p38MAPK in the asthma model group was significantly higher than that of the normal control group;The levels of these proteins in the Jingfang Baidu lower,high dose group and dexamethasone positive control group were lower than those in the asthma group.The expression and location of NF-?Bp65 in lung tissue of mice were detected by immunohistochemistry and Western blot.The results showed that the expression of NF-?B p65 was increased in the asthma model group compared with the normal control group;However,the NF-?Bp65 was significantly decreased in the high-dose group and the dexamethasone-positive control group compared with the Asthma model group(p?0.05).Jingfang Baidu low dose group showed no significant difference compared with asthma model group.Conclusions:JingFangBaiDu Powder can play a role in alleviating airway inflammation in asthma,and its possible mechanism may be related to inhibition of p38MAPK phosphorylation and inhibition of NF-?Bp65 activation. |
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