穿心莲内酯磺化物对支气管哮喘小鼠肺泡灌洗液炎性细胞、白细胞介素-12及白细胞介素-13表达的影响

Effect of Andrographolide on Expressions of Inflammatory Cells,Interleukin-12 and Interleukin-13 in Bronchoalveolar Larage Fluid of Asthmatic Mice Model

目的探讨穿心莲内酯磺化物对哮喘小鼠肺泡灌洗液(BALF)内炎性细胞、IL-12及IL-13表达的影响。方法 BALB/c小鼠随机分为哮喘组、穿心莲内酯磺化物干预组和对照组。以卵清蛋白(OVA)致敏激发建立哮喘小鼠模型。计数BALF炎性细胞总数及分类,双抗体夹心ELISA法测定BALF中IL-12、IL-13水平,实时荧光定量PCR检测肺组织中IL-12 mRNA、IL-13 mRNA相对表达量。结果哮喘组、穿心莲内酯磺化物干预组、对照组BALF中白细胞总数分别为(203.09±43.60)×104L-1、(123.90±35.10)×104 L-1、(20.69±3.68)×104 L-1;嗜酸性粒细胞(EOS)数分别为(160.78±31.62)×104 L-1、(67.44±20.00)×104 L-1、(1.72±0.68)×104 L-1;巨噬细胞数分别为(33.85±2.29)×104 L-1、(14.12±2.10)×104 L-1、(17.24±1.97)×104 L-1;BALF中IL-12水平分别为(60.45±7.91)ng.L-1、(92.27±18.67)ng.L-1、(278.48±36.78)ng.L-1;BALF中IL-13水平分别为(570.20±62.98)ng.L-1、(299.71±31.64)ng.L-1、(230.22±27.83)ng.L-1;肺组织IL-12 p40 mRNA相对表达量分别为0.43±0.24、0.93±0.28、1.04±0.28;IL-13 mRNA相对表达量分别为44.80±9.51、11.27±5.47、1.07±0.47。上述指标,三组间比较差异均有统计学意义(Pa<0.01)。结论穿心莲内酯磺化物可抑制哮喘小鼠炎性细胞浸润和IL-13表达,促进IL-12表达拮抗哮喘呼吸道炎症。

Objective To explore the effects of andrographolide on expressions of inflammatory cells,IL^-12 and IL^-13 in bronchoalveolar lavage fluid（BALF） of asthmatic mice. Methods BALB/c mice were randomly divided into asthmatic group,andrographolide intervention group and control group.Mice were sensitized and challenged with ovalbumin（OVA） to develop asthmatic model.The total and diffe-rential numbers of inflammatory cells in BALF were counted.The concentrations of IL^-12 and IL^-13 in BALF were detected by Double-antibody sandwich enzyme-linked immune assay（ELISA）.The relative expressions of IL^-12 mRNA and IL^-13 mRNA of lung tissue were detected by real time fluorescent quantitative PCR. Results In asthmatic group,andrographolide intervention group and control group,the total cell counts were（203.09±43.60）×10^4 L^-1,（123.90±35.10）×10^4 L^-1 and（20.69±3.68）×10^4 L^-1,respectively;the eosinophil（EOS） counts were（160.78±31.62）×10^4 L^-1,（67.44±20.00）×10^4 L^-1 and（1.72±0.68）×10^4 L^-1,respectively;the macrophages（Mac） counts were（33.85±2.29）×10^4 L^-1,（14.12±2.10）×10^4 L^-1,（17.24±1.97）×10^4 L^-1,respectively;the concentrations of IL^-12 in BALF were（60.45±7.91） ng·L^-1,（92.27±18.67） ng·L^-1 and（278.48±36.78） ng·L^-1,respectively;the IL^-13 in BALF were（570.20±62.98） ng·L^-1,（299.71±31.64） ng·L^-1 and（230.22±27.83） ng·L^-1,respectively;the relative expressions of IL^-12 p40 mRNA in lung tissue were 0.43±0.24,0.93±0.28 and 1.04±0.28,respectively;the relative expressions of IL^-13 mRNA in lung tissue were 44.80±9.51,11.27±5.47 and 1.07±0.47,respectively.There were statistically significant differences in these indexes among the 3 groups（Pa0.01）. Conclusions Andrographolide can significantly antagonize airway inflammatory by reducing various inflammatory cells infiltration,IL^-13 expression,and promoting IL^-12 expression.