小儿柴桂退热口服液对脂多糖致肺损伤模型小鼠的作用及机制

Effect and Mechanism of Xiaoer Chaigui Antipyretic Oral Liquid on Lipopolysaccharide-Induced Lung Injury Model Mice

目的探讨小儿柴桂退热口服液对脂多糖诱导的肺损伤模型小鼠的作用及机制。方法将60只雄性KM小鼠随机分为空白对照组(A组,生理盐水),模型对照组(B组,生理盐水),小儿柴桂退热口服液低、中、高剂量组(C_(1)组、C_(2)组、C_(3)组,8,16,32 mg/kg),阳性对照组(D组,5 mg/kg连花清瘟胶囊),各10只。除A组外,其余各组小鼠均一次性尾静脉注射脂多糖5 mg/kg,以复制肺损伤模型,建模成功后均灌胃相应药物或生理盐水,每天1次,连续3 d。检测小鼠肺组织干/湿质量比;苏木素-伊红染色,显微镜下观察小鼠肺组织的病理形态;采用酶联免疫吸附(ELISA)法检测小鼠血浆中白细胞介素1β(IL-1β)、肿瘤坏死因子-α(TNF-α)的水平;采用全自动血细胞分析仪检测小鼠的血常规指标;采用免疫印迹(Western blot)法检测小鼠肺组织中核转录因子-κB(NF-κB)、核苷酸结合寡聚化结构域(NOD)样受体热蛋白结构域相关蛋白3(NLRP3)和IL-1β的蛋白表达水平。结果与B组比较,C_(2)组、C_(3)组小鼠肺组织干/湿质量比及外周血白细胞计数(WBC)、血小板计数(PLT)、淋巴细胞(LYMPH)均显著升高(P<0.05),外周血红细胞比容(HCT)、中性粒细胞计数比例(NEUT%)及肺组织中NLRP3蛋白表达水平均显著降低(P<0.05);C_(1)组、C_(2)组、C_(3)组小鼠血浆中IL-1β和TNF-α及肺组织中IL-1β和NF-κB蛋白表达水平均显著降低(P<0.05)。结论小儿柴桂退热口服液可减轻脂多糖诱导的肺损伤模型小鼠肺组织的炎性损伤程度,其作用机制可能与调控NF-κB/NLRP3信号通路有关。

Objective To investigate the effect and mechanism of Xiaoer Chaigui Antipyretic Oral Liquid on the lipopolysaccharide(LPS)-induced lung injury model mice.Methods A total of 60 male KM mice were randomly divided into the blank control group(group A,treated with normal saline),the model control group(group B,treated with normal saline),the Xiaoer Chaigui Antipyretic Oral Liquid low-,medium-and high-dose groups(groups C_(1),C_(2) and C_(3),treated with 8,16,32 mg/kg of Xiaoer Chaigui Antipyretic Oral Liquid)and the positive control group(group D,treated with 5 mg/kg of Lianhua Qingwen Capsules),with 10 mice in each group.Except for group A,all mice in the other groups were injected with 5 mg/kg of LPS once via the tail vein to replicate the lung injury model.After the successful modeling,the mice were given the corresponding drugs or normal saline by gavage once a day for 3 d continuously.The dry/wet weight ratio of lung tissue was detected.The pathological morphology of lung tissue under the microscope was observed by the hematoxylin-eosin(HE)staining method.The interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)levels in plasma were detected by the enzyme-linked immunosorbent assay(ELISA).The routine blood indexes of mice were detected by the automatic blood cell analyzer.The expression levels of nuclear transcription factor-κB(NF-κB),nucleotide-binding oligomerization domain(NOD)-like receptor thermal protein domain associated protein 3(NLRP3)and IL-1β proteins in lung tissue were detected by the Western blot.Results Compared with those in the group B,the dry/wet weight ratio of lung tissue,white blood cell count(WBC),platelet count(PLT)and lymphocyte(LYMPH)of peripheral blood in the groups C_(2)and C_(3)were significantly higher(P<0.05),the hematocrit(HCT),neutrophil proportion(NEUT%)of peripheral blood and the expression level of NLRP3 protein in lung tissue in the groups C_(2)and C_(3) were significantly lower(P<0.05).Compared with those in the group B,the expression levels of IL-1β,TNF-α proteins in plasma and IL-1β,NF-κB proteins in lung tissue in the groups C_(1),C_(2) and C_(3)were significantly lower(P<0.05).Conclusion Xiaoer Chaigui Antipyretic Oral Liquid can reduce the degree of inflammatory injury of lung tissue in LPS-induced lung injury model mice,and its mechanism may be related to the regulation of NF-κB/NLRP3 signal pathway.