清热润燥口服液对哮喘小鼠ILC2s上下游细胞因子的影响

Effect of Qingre Runzao oral liquid on chemokines and ILC2s and their upstream and downstream cytokines in asthmatic mice

目的通过清热润燥口服液干预卵清蛋白(OVA)致敏的哮喘小鼠,观察其对哮喘小鼠Ⅱ型固有淋巴细胞(ILC2s)数量及其上下游细胞因子的影响。方法将BALB/c小鼠(雌性)分为空白对照组、模型对照组、阳性对照组(地塞米松组,1 mg/(kg·d))、清热润燥口服液低、高(4.32、8.64 g/(kg·d))剂量组,共40只。使用OVA为致敏剂诱导哮喘小鼠模型,HE染色观察小鼠肺组织的病理改变;酶联免疫吸附(ELISA)测定检测小鼠肺泡灌洗液(BALF)中ILC2s上下游细胞因子Eotaxin、IL-25、IL-33、TSLP、IL-4、IL-5及IL-6含量;免疫组化法检测肺组织ILC2s上下游细胞因子IL-33和IL-5的表达;流式细胞术和免疫双荧光染色检测肺组织中ILC2s的数量及生成。结果与空白对照组比较,模型对照组小鼠出现肺组织炎性细胞浸润和支气管壁增厚的病理变化,BALF中ILC2s上下游细胞因子Eotaxin、IL-25、IL-33、TSLP、IL-4、IL-5及IL-6的含量升高(P<0.01),肺组织中ILC2s上下游细胞因子IL-33及IL-5的表达升高(P<0.01)、ILC2s的数量及生成增多(P<0.01)。与模型对照组比较,清热润燥口服液低高剂量明显改善哮喘小鼠肺组织病理损伤,降低BALF中ILC2s上下游细胞因子表达(P<0.01)、下调肺组织中ILC2s上下游细胞因子IL-33及IL-5的表达(P<0.05,P<0.01)和减少ILC2s的数量及生成(P<0.01)。结论清热润燥口服液可以降低OVA诱导的哮喘小鼠ILC2s的数量及其上下游细胞因子和下调肺组织中IL-33及IL-5的表达,进而起到防治哮喘的作用。

Objective To observe the effects of Qingre Runzao oral liquid on the numbers of type Ⅱ intrinsic lymphocytes( ILC2 s) and their upstream and downstream cytokines in asthmatic mice sensitized by ovalbumin. Methods Female BALB/c mice were divided into the blank control group, model control group, positive control group( dexamethasone group,1 mg/( kg·d)) and low-and high-dose Qingre Runzao oral liquid groups( 4. 32 and 8. 64 g/( kg·d)). The asthma mouse model was induced using ovalbumin as a sensitizer,and pathological changes in lung tissue were observed by hematoxylin and eosin staining. Levels of the upstream and downstream cytokines of ILC2 s Eotaxin,interleukin( IL)-25,IL-33,thymic stromal lymphopoietin( TSLP),IL-4,IL-5 and IL-6 were detected in mouse bronchoalveolar lavage fluid( BALF) using enzyme-linked immunosorbent assay. The expression of IL-33 and IL-5 in lung tissue was detected by immunohistochemical method. Flow cytometry and immunofluorescence staining were used to assess the number and production of ILC2 s in lung tissue. Results Compared with the blank control group,the model control group showed pathological changes of inflammatory cell infiltration in the lung tissue and bronchial wall thickening. Furthermore,levels of the upstream and downstream cytokines Eotaxin,IL-25,IL-33,TSLP,IL-4,IL-5 and IL-6 were increased in BALF( P<0. 01). The expression of IL-33 and IL-5 was increased in lung tissue( P<0. 01),and the number and production of ILC2 s was also increased( P<0. 01). Compared with the model control group,both the low and high doses of Qingre Runzao oral liquid significantly improved the pathological injury to lung tissue in asthmatic mice,decreased the expression of upstream and downstream cytokines of ILC2 s in BALF( P < 0. 01),downregulated the expression of IL-33 and IL-5 in lung tissue( P<0. 05,P<0. 01),and decreased the number and production of ILC2 s( P<0. 01). Conclusions Qingre Runzao oral liquid can reduce the numbers of ILC2 s and their upstream and downstream cytokines in ovalbumin-induced asthmatic mice.It can also downregulate the expression of IL-33 and IL-5 in lung tissue,thus playing a role in the prevention and treatment of asthma.