黄连素对COPD大鼠气道炎症和粘液高分泌的调控作用研究

Effect of Berberine on airway inflammation and mucus hypersecretion in mice with chronic obstructive pulmonary disease and its molecular mechanism

目的探讨黄连素对COPD大鼠气道炎症和粘液高分泌的影响及研究其相关分子机制。方法清洁级雄性Wistar大鼠随机分为生理盐水对照组、COPD模型组和黄连素组各10只,通过熏香烟加气管注内毒素方法建立大鼠COPD模型。采集支气管肺泡灌洗液用ELISA法检测中性粒细胞弹性蛋白酶(neutrophil elastase,NE)、IL-10含量。采集支气管-肺组织标本用免疫组织化学法(SP法)检测NF-κB、粘蛋白Muc5ac表达,采用逆转录-聚合酶链反应(RT-PCR)检测Muc5ac mRNA表达。结果 COPD模型组大鼠BALF细胞总数、中性粒细胞数(PMN)、巨噬细胞数(AM)、NE较正常对照组显著升高(P<0.01),IL-10水平显著下降(P<0.01)。黄连素组BALF细胞总数、PMN、AM较COPD组显著降低(P<0.01),L-10水平升则高(P<0.05)。COPD模型组大鼠气道黏膜上皮NF-κB、Muc5ac mRN及蛋白表达较正常对照组显著升高(P<0.01)。黄连素组气道黏膜上皮NF-κB、Muc5ac mRN及蛋白表达较COPD组显著下降(P<0.01)。结论黄连素可负向调节气道炎症和粘液高分泌,其机制可能与黄连素抑制NE、NF-κB炎症因子,促进抗炎因子IL-10,下调粘蛋白Muc5ac表达有关。

Objective To investigate the effect of Berberine on airway inflammation and mucus hypersecretion in mice with chronic obstructive pulmonary disease and its molecular mechanism.Methods Thirty male and healthy SD mice were randomly divided into three groups,with ten mice in each group,include NS control,COPD and berberine groups.Chronic obstructive pulmonary disease mice models were established by passive cigarette smoking plus intratracheal instillation of lipopolysaccharide.Bronchoalveolar lavage fluid（BALF）and lung tissue sample were collected.Total and differential cell counts in BALF were measured,and neutrophil elastase（NE）,interleukin-10（IL-10）were detected with enzyme linked immunosorbent assay.Meanwhile,nuclear factor-kappaB and Muc5ac protein were detected with immunohistochemistry,and muc5ac mRNA by RT-PCR.Results Total cell counts,PMN counts,AM counts and level of NE in BALF of COPD model group were significantly increased than those of control group（P0.01）,however the level of IL-10 was significantly decreased（P0.01）.Total cell counts,PMN counts,AM counts and level of NE in BALF of Berberine treated group were significantly decreased than those of COPD group（P0.01）,the level of IL-10 was increased（P0.05）.The expressions of NF-κB,Muc5ac mRN and its protein in COPD model group were significantly increased than those in control group（P0.01）.The expressions of NF-κB、Muc5ac mRN and its protein in Berberine treated model group were significantly decreased than those in COPD group（P0.01）.Conclusion Berberine may negative regulate airway inflammation and mucus hypersecretion and its mechanism may be related to inhibit inflammatory cytokines NE,NF-κB,promote anti-inflammatory factor IL-10 and reduce the expression of mucin Muc5ac.