温肾方对哮喘小鼠ILC2相关分子表达的影响

Effect of Wenshen Formula on ILC2-related molecules expression in asthmatic mice

目的研究温肾方对哮喘小鼠2型固有淋巴样细胞(ILC2)相关分子表达的影响。方法 30只BALB/c小鼠随机分为正常对照组(NC)、IL-33组、西药组(PRE)、温肾方组(WSF)、中西医结合组(CO),每组6只。除正常对照组外,其余4组用IL-33(每40μl含0.4μg)滴鼻,连续6天,第7天开始隔天滴鼻,持续1周。西药组(醋酸泼尼松1.125 g/kg)、温肾方组(温肾方9 g/kg)和中西医结合组(温肾方加醋酸泼尼松)均在滴鼻前1 h灌胃给予相应药物干预,共15天。采用HE染色观察各组肺组织的形态和炎症细胞的浸润情况;以ELISA法检测肺泡灌洗液中IL-4、IL-5、IL-13细胞因子的表达;采用荧光定量PCR检测肺组织中RORα的表达情况。结果①与正常对照组比较,IL-33组小鼠肺组织黏膜皱襞增厚,炎性细胞明显浸润,温肾方组、西药组和中西医结合组均可明显降低炎性细胞的浸润。②与正常对照组比较,IL-33组小鼠肺泡灌洗液中IL-4、IL-5、IL-13的表达水平明显升高(P<0.001),温肾方组、西药组和中西医结合组均可以明显下调小鼠BALF中IL-4、IL-5、IL-13的表达水平,与IL-33组比较差异有统计学意义(P<0.01)。③与正常对照组比较,IL-33组可明显上调小鼠肺组织中ILC2的RORαmRNA的表达,差异有统计学意义(P<0.001);温肾方组、西药组和中西医结合组均可以明显下调小鼠肺组织RORα的mRNA表达,与IL-33组比较差异有统计学意义(P<0.05)。结论温肾方能减轻小鼠气道炎症细胞的浸润,缓解哮喘炎症;并可下调ILC2分泌相关的细胞因子IL-4、IL-5、IL-13的表达和ILC2特异性转录因子RORαmRNA的表达。

Objective To study the effect of Wenshen Formula on the expression of group 2 innate lymphoid cells(ILC2)-related molecules in asthmatic mice. Methods Thirty BALB/c mice were randomly assigned into the normal control(NC) group, the IL-33 group, the Western medicine(PRE) group, the Wenshen Formula(WSF) group and the combination(CO) group, 6 mice in each group. Except the normal control group, rmIL-33 nasal drops(0.4 μg/40 μl) were given to the other 4 groups for 6 consecutive days and starting at day 7, the drug was administered once every other day for 1 week. Drug invention was given by gavage for 15 days to the PRE group(prednisone acetate 1.125 g/kg), WSF group(Wenshen Formula 9 g/kg) and CO group(Wenshen Formula + prednisone acetate) at 1 hour before nasal administration. Lung tissue was stained with hematoxylin-eosin(HE) to observe morphology of the tissue and infiltration of inflammatory cells. ELISA was used to detect the expression of IL-4, IL-5 and IL-13 in bronchoalveolar lavage fluid(BALF). Fluorescence quantitative PCR was used to detect the expression of RORα in lung tissue. Results ①Compared to the NC group, the mucosal folds of lung tissue were thicker and infiltration of inflammatory cells was more obvious in the IL-33 group. Infiltration of inflammatory cells was reduced significantly in the WSF, PRE and CO groups. ②Compared to the NC group, the expression levels of IL-4, IL-5 and IL-13 in BALF of mice increased significantly in the IL-33 group(P<0.001). The expression levels of IL-4, IL-5 and IL-13 in BALF were reduced in the WSF, PRE and CO groups, with significant differences compared to the IL-33 group(P<0.01). ③Compared to the NC group, the expression of ILC2 RORα mRNA in lung tissue was increased significantly in the IL-33 group(P<0.001). The expression of RORα mRNA in lung tissue was downregulated in the WSF,PRE and CO groups,with statistically significant differences compared to the IL-33 group( P<0.05).Conclusion Wenshen Formula reduces infiltration of the airway inflammatory cells and alleviates asthmatic inflammation in mice. It down-regulates the expression of IL-4,IL-5 and IL-13 and of RORα mRNA.