屈头鸡果对哮喘小鼠气道炎症细胞及肺组织NF-κBp65、TGF-β1的调节作用研究
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摘要
目的观察在屈头鸡果的干预作用下,支气管哮喘小鼠支气管肺泡灌洗液(BALF)白细胞(WBC)、嗜酸粒细胞(EOS)数量的变化及其肺组织核转录因子κBp65(NF-κBp65)、转化生长因子β1(TGF-β1)表达水平。方法SPF级BALB/c雌性小鼠50只,随机分为5组,每组10只。A组为正常对照组(腹腔注射生理盐水); B组为哮喘模型组(腹腔注射致敏液); C组为屈头鸡果治疗剂量组(腹腔注射致敏液+灌胃屈头鸡果浆2. 16 g/kg); D组为屈头鸡果中毒剂量组(腹腔注射致敏液+灌胃屈头鸡果浆10. 8 g/kg); E组为地塞米松治疗组(腹腔注射致敏液+地塞米松2 mg/kg)。在末次激发结束后采集其BALF和肺组织,观察镜下各组小鼠BALF中WBC总数与EOS计数;采用蛋白印迹杂交(Western-blot)法测定各组小鼠肺组织NF-κBp65、TGF-β1表达水平。结果 A组小鼠BALF中WBC总数、EOS计数以及肺组织NF-κBp65、TGF-β1表达水平均显著低于B组、C组、D组和E组,差异有显著统计学意义(P <0. 01);C组、D组小鼠BALF中WBC总数、EOS计数显著低于B组(P <0. 05); C组、D组小鼠肺组织NF-κBp65、TGF-β1表达水平亦显著低于B组(P <0. 05); D组有部分小鼠死亡。结论屈头鸡果可调节哮喘小鼠气道中炎症细胞的数量以及肺组织NF-κBp65、TGF-β1的表达水平,从而抑制哮喘气道炎症;但屈头鸡果具有一定毒性,不可过量服用。
Objective To observe the changes of inflammatory cells( WBC and EOS) count in bronchoalveolar lavage fluid( BALF) and NF-κBp65,TGF-β1 expression level in lung tissue in asthmatic mice intervened by Capparis versicolor griff. Methods 50 SPF class BALB/c female mice were randomly divided into 5 groups,10 mice in each group. The group A was the normal control group( intraperitoneal injection of normal saline),the group B was the asthmatic model group( intraperitoneal sensitizing solution),and the group C was the group of Capparis versicolor griff treatment dose( intraperitoneal sensitizing solution + gastric perfusion capparis versicolor griff 2. 16 g/kg); the group D was the group of Capparis versicolor Griff poisoning dose( intraperitoneal sensitizing solution + gastric perfusion capparis versicolor griff 10. 8 g/kg),and the group E was the Dexamethasone treatment group( intraperitoneal sensitizing solution + dexamethasone). To collect the BALF and the lung tissue of the mice at the end of the last challenge,and observe WBC count and EOS count in BALF; observe NF-κBp65 and TGF-β1 expression in lung tissue assay by Western-blot. Results The total number of WBC,EOS count in BALF and the expression of NF-κBp65 and TGF-β1 of group A were significantly lower than those of group B,C,D and E. The difference was statistically significant( P < 0. 01). The counts of WBC and EOS in BALF in group C and group D were lower than group B with significant difference( P < 0. 05); The NF-κBp65 and TGF-β1 expression level in group C and group D were also lower than group B( P < 0. 05). Some mice died in group D. Conclusion Capparis versicolor griff can reduce the counts of inflammatory cells in airway; and lower the expression levels of NF-κBp65 and TGF-β1 in lung tissue. Thus capparis versicolor griff can suppress the airway inflammation in asthma. But capparis versicolor griff has certain toxicity and should not be taken overdose.
Objective To observe the changes of inflammatory cells( WBC and EOS) count in bronchoalveolar lavage fluid( BALF) and NF-κBp65,TGF-β1 expression level in lung tissue in asthmatic mice intervened by Capparis versicolor griff. Methods 50 SPF class BALB/c female mice were randomly divided into 5 groups,10 mice in each group. The group A was the normal control group( intraperitoneal injection of normal saline),the group B was the asthmatic model group( intraperitoneal sensitizing solution),and the group C was the group of Capparis versicolor griff treatment dose( intraperitoneal sensitizing solution + gastric perfusion capparis versicolor griff 2. 16 g/kg); the group D was the group of Capparis versicolor Griff poisoning dose( intraperitoneal sensitizing solution + gastric perfusion capparis versicolor griff 10. 8 g/kg),and the group E was the Dexamethasone treatment group( intraperitoneal sensitizing solution + dexamethasone). To collect the BALF and the lung tissue of the mice at the end of the last challenge,and observe WBC count and EOS count in BALF; observe NF-κBp65 and TGF-β1 expression in lung tissue assay by Western-blot. Results The total number of WBC,EOS count in BALF and the expression of NF-κBp65 and TGF-β1 of group A were significantly lower than those of group B,C,D and E. The difference was statistically significant( P < 0. 01). The counts of WBC and EOS in BALF in group C and group D were lower than group B with significant difference( P < 0. 05); The NF-κBp65 and TGF-β1 expression level in group C and group D were also lower than group B( P < 0. 05). Some mice died in group D. Conclusion Capparis versicolor griff can reduce the counts of inflammatory cells in airway; and lower the expression levels of NF-κBp65 and TGF-β1 in lung tissue. Thus capparis versicolor griff can suppress the airway inflammation in asthma. But capparis versicolor griff has certain toxicity and should not be taken overdose.
引文
[1]Nathan RA,Thompson PJ,Price D,et al.Taking Aim at Asthma Around the World:Global Results of the Asthma Insight and Management Survey in the Asia-Pacific Region,Latin America,Europe,Canada,and the United States[J].J Allergy Clin Immunol Pract,2015,3(5):734-742.
[2]《全国中草药汇编》编写组.全国中草药汇编下册[M].北京:人民卫生出版社,1986:376.
[3]Bel EH,Ten Brinke A.New Anti-Eosinophil Drugs for asthma and COPD:targeting the trait[J].Chest,2017,152(6):1276-1282.
[4]Casciano J,Krishnan JA,Small MB,et al.Burden of asthma with elevated blood eosinophil levels[J].BMC Pulm Med,2016,16(1):100.
[5]GM Walsh.Antagonism of cytokine-induced eosinophil accumulation in asthma[J].Front Pharmacol,2012,3(3):197.
[6]Ullmann N,Bossley CJ,Fleming L,et al.Blood eosinophil counts rarely reflect airway eosinophilia in children with severe asthma[J].Allergy,2013,68(3):402-406.
[7]Christman JW,Sadikot RT,Blackwell TS,et al.The role of nuclear factor-κB in pulmonary diseases[J].Chest,2000,117(5):1482-1487.
[8]Payne AS,Freishtat RJ.Conserved steroid hormone homology converges on NF-κB to modulate inflammation in asthma[J].J Investig Med,2012,60(1):13-17.
[9]Manuyakorn W,Kamchaisatian W,Atamasirikul K,et al.Serum TGF-β1 in Atopic Asthma[J].Asian Pac J Allergy Immunol,2008,26(4):185-189.
[10]Yim RP,Koumbourlis AC.Steroid-resistant asthma[J].Paediatr Respir Rev,2012,13(3):172-176.
[11]Lefebvre P,Duh MS,Lafeuille MH,et al.Burden of systemic glucocorticoid-related complications in severe asthma[J].Curr Med Res Opin,2017,33(1):57-65.
[12]覃冠德,黄正志.屈头鸡果急性中毒34例救治分析[J].实用医学杂志,2012,28(6):1029.
[2]《全国中草药汇编》编写组.全国中草药汇编下册[M].北京:人民卫生出版社,1986:376.
[3]Bel EH,Ten Brinke A.New Anti-Eosinophil Drugs for asthma and COPD:targeting the trait[J].Chest,2017,152(6):1276-1282.
[4]Casciano J,Krishnan JA,Small MB,et al.Burden of asthma with elevated blood eosinophil levels[J].BMC Pulm Med,2016,16(1):100.
[5]GM Walsh.Antagonism of cytokine-induced eosinophil accumulation in asthma[J].Front Pharmacol,2012,3(3):197.
[6]Ullmann N,Bossley CJ,Fleming L,et al.Blood eosinophil counts rarely reflect airway eosinophilia in children with severe asthma[J].Allergy,2013,68(3):402-406.
[7]Christman JW,Sadikot RT,Blackwell TS,et al.The role of nuclear factor-κB in pulmonary diseases[J].Chest,2000,117(5):1482-1487.
[8]Payne AS,Freishtat RJ.Conserved steroid hormone homology converges on NF-κB to modulate inflammation in asthma[J].J Investig Med,2012,60(1):13-17.
[9]Manuyakorn W,Kamchaisatian W,Atamasirikul K,et al.Serum TGF-β1 in Atopic Asthma[J].Asian Pac J Allergy Immunol,2008,26(4):185-189.
[10]Yim RP,Koumbourlis AC.Steroid-resistant asthma[J].Paediatr Respir Rev,2012,13(3):172-176.
[11]Lefebvre P,Duh MS,Lafeuille MH,et al.Burden of systemic glucocorticoid-related complications in severe asthma[J].Curr Med Res Opin,2017,33(1):57-65.
[12]覃冠德,黄正志.屈头鸡果急性中毒34例救治分析[J].实用医学杂志,2012,28(6):1029.