冬凌草甲素对急性哮喘小鼠Th17/Treg细胞平衡的影响

Influence of oridonin on Th17/Treg cell balance in acute asthmatic mice

目的探讨冬凌草甲素对急性哮喘小鼠辅助性T细胞17/调节性T淋巴细胞(Th17/Treg)平衡的影响及可能的作用机制。方法将雌性清洁级BALB/c小鼠100只随机分为4组,分别为正常组、模型组、冬凌草甲素10 mg/kg干预组(低剂量干预组)和冬凌草甲素20 mg/kg干预组(高剂量干预组)四组,每组各25只。建立鸡卵清白蛋白(OVA)诱导的急性哮喘模型,造模成功后,低剂量干预组腹腔注射冬凌草甲素10 mg/kg,高剂量干预组腹腔注射冬凌草甲素20 mg/kg。正常组以PBS代替OVA进行致敏、生理盐水代替OVA激发。计数支气管肺泡灌洗液(BALF)细胞总数、嗜酸粒细胞百分比;采用HE染色对炎症细胞浸润情况及肺组织病理学改变进行观察;ELISA方法检测气道炎症相关细胞因子IL-17、IL-22、IL-10、IL-35的表达水平;AB-PAS染色后观察粘液物质分泌及气道上皮杯状细胞增生的情况。结果与正常组相比,模型组、干预组的促炎因子表达水平(IL-17及IL-22)、气道上皮杯状细胞和粘液物质阳性相对着色面积均明显升高(P<0.05),抑炎因子(IL-10、IL-35)表达水平均明显减低(P<0.05),差异有统计学意义;与模型组相比,干预组的促炎因子表达水平(IL-17及IL-22)、气道上皮杯状细胞和粘液物质阳性相对着色面积均明显降低(P<0.05),抑炎因子IL-10、IL-35表达水平明显升高(P<0.05);与低剂量干预组比较,高剂量干预组的促炎因子表达水平(IL-17及IL-22)、气道上皮杯状细胞和粘液物质阳性相对着色面积均降低(P<0.05),抑炎因子IL-10、IL-35表达水平均升高(P<0.05)。结论冬凌草甲素能减轻急性哮喘小鼠气道炎症反应,可能是通过抑制促炎因IL-17、IL-22分泌及促进抑炎因子IL-10、IL-35分泌,从而导致Th17/Treg细胞失衡来实现的。

Objective To investigate the effect of oridonin on the balance of T helper 17/regulatory T lymphocytes(Th17/Treg)in acute asthma mice and its possible mechanism of action.Methods Female clean-grade BALB/c mice were randomly divided into 4 groups,namely normal group,model group,Rubescensine 10 mg/kg intervention group(low-dose intervention group),and Rubescensine 20 mg/kg intervention group(high-dose intervention group).A chicken ovalbumin(OVA)-induced acute asthma model was established.After successful modeling,the low-dose intervention group was injected with oridonin 10 mg/kg intraperitoneally,and the high-dose intervention group was intraperitoneally injected with oridonin 20 mg/kg.The normal group was sensitized with PBS instead of OVA,and challenged with normal saline instead of OVA.The total number of cells in bronchoalveolar lavage fluid(BALF)and the percentage of eosinophils were counted.HE staining was used to observe the infiltration of inflammatory cells and pathological changes in lung tissue.ELISA was used to detect airway inflammation-related cytokines IL-17,IL-22,IL-10,and IL-35.After AB-PAS staining,the secretion of mucous substances and the hyperplasia of airway epithelial goblet cells were observed.Results Compared with the normal group,the expression levels of pro-inflammatory factors(IL-17 and IL-22),the airway epithelial goblet cells and the positive relative staining area ofmucous substances in the model group and the intervention group were significantly increased(P<0.05),the expression levels of anti-inflammatory factors(IL-10,IL-35)were significantly decreased(P<0.05),and the difference was statistically significant.Compared with the model group,the expression levels of pro-inflammatory factors(IL-17 and IL-22)in the intervention group,airway epithelial goblet cells and the positive relative staining area of mucous substances were significantly decreased(P<0.05),and the expression levels of anti-inflammatory factors IL-10 and IL-35 were significantly increased(P<0.05).Compared with the low dose intervention group,the expression levels of pro-inflammatory factors(IL-17 and IL-22),the positive relative staining area of airway epithelial goblet cells and mucous substances in the high-dose intervention group were decreased(P<0.05),and the expression levels of anti-inflammatory factor IL-10 and IL-35 were increased(P<0.05).Conclusions Rubescensine could reduce the airway inflammatory response of acute asthma mice,which may be by inhibiting the secretion of pro-inflammatory factors IL-17 and IL-22 and promoting the secretion of anti-inflammatory factors IL-10 and IL-35,thereby leading to Th17/Treg cell imbalance.