连花清瘟对细颗粒物致大鼠肺部炎性损伤的拮抗作用研究

摘要

目的：探讨细颗粒物（ PM2.5）急性暴露对大鼠肺部炎性损伤的作用，及低、中、高剂量连花清瘟对肺部炎性损伤的拮抗作用。方法2013年6—12月选取48只健康成年Wistar大鼠，采用随机数字表法分为空白对照组、0.9%氯化钠溶液对照组、PM2.5染尘组及低、中、高剂量连花清瘟组，每组8只。由石家庄市环境监测中心提供PM2.5，制备PM2.5悬液。空白对照组大鼠无任何干预措施；0.9%氯化钠溶液对照组大鼠给予气管滴注0.9%氯化钠溶液（1 ml/kg）；PM2.5染尘组大鼠给予气管滴注PM2.5悬液1 ml/kg（7.5 mg/kg）；低、中、高剂量连花清瘟组大鼠首先分别连续灌胃给予低（2 g/kg）、中（4 g/kg）、高（8 g/kg）剂量连花清瘟溶液10 ml/kg，共4 d，第4天灌胃给药后分别气管滴注 PM2.5悬液1 ml/kg（7.5 mg/kg），24 h 后处死。光镜下观察肺组织病理形态变化，酶联免疫吸附试验（ELISA）法测定各组大鼠支气管肺泡灌洗液（BALF）及血清中白介素（IL）1、IL-6、肿瘤坏死因子α（TNF-α）水平。结果空白对照组及0.9%氯化钠溶液对照组大鼠肺组织病理切片光镜下未见异常表现；PM2.5染尘组大鼠肺组织病理切片光镜下可见炎性细胞渗出，肺间质纤维组织增生，间质水肿；低、中、高剂量连花清瘟组大鼠肺组织病理切片光镜下可见随连花清瘟剂量增加，肺泡腔内炎性细胞渗出逐渐减轻。0.9%氯化钠溶液对照组大鼠BALF及血清中IL-1、IL-6、TNF-α水平与空白对照组比较，差异均无统计学意义（ P>0.05）；PM2.5染尘组大鼠BALF中IL-1水平较0.9%氯化钠溶液对照组升高，血清中IL-1水平、BALF及血清中IL-6、TNF-α水平较空白对照组和0.9%氯化钠溶液对照组升高（ P<0.05）；高剂量连花清瘟组大鼠BALF中IL-1水平较空白对照组、0.9%氯化钠溶液对照组、PM2.5染尘组、低剂量连花清瘟组、中剂量连花清瘟组降低，BALF和血清中IL-6水平较空白对照组、0.9%氯化钠溶液对照组、PM2.5染尘组、低剂量连花清瘟组降低，BALF中TNF-α水平和血清中IL-1水平较PM2.5染尘组、低剂量连花清瘟组、中剂量连花清瘟组降低，血清中TNF-α水平较PM2.5染尘组、低剂量连花清瘟组降低（P<0.05）。结论PM2.5急性暴露可以导致大鼠肺部炎性损伤，连花清瘟对肺部炎性损伤有拮抗作用。%Objective To investigate the pulmonary inflammatory injury in rats induced by acute exposure to PM2. 5 and to explore the antagonistic effect of low,medium,and high dose of Lianhuaqingwen to the inflammatory injury. Methods From June to December in 2013,selected 48 healthy adult Wistar rats. Using random number table method,divided them into control group,sodium chloride group,PM2. 5 group,low-dose group,medium-dose group and high-dose group,with 8 rats in each group. Airborne fine particulate matters were provided by Shijiazhuang Environmental Monitoring Center,and PM2. 5 suspension liquid was prepared. Control group was not given any intervention;sodium chloride group was given trachea instillation of 0. 9%sodium chloride solution( 1 ml/kg );PM2. 5 group was given trachea instillation of 1 ml/kg ( 7. 5 mg/kg ) PM2. 5 suspension liquid;low-dose,medium-dose and high -dose groups were respectively given gavage with 2 g/kg,4 g/kg and 8 g/kg Lianhuaqingwen solution of 10 ml/kg for 4 days,and the three groups were given trachea instillation of 1 ml/kg(7. 5 mg/kg) PM2. 5 suspension liquid on the fourth day and were killed 24 hours later. Pathological morphologic changes were observed under light microscope,and ELISA was employed to examine the levels of IL-1,IL-6 and TNF-αin BALF and serum. Results In control group and sodium chloride group, no abnormal manifestations in pulmonary tissue slices were observed under light microscope;in PM2. 5 group,exudation of inflammatory cells,interstitial proliferation of fibrous tissue and interstitial edema were noted in pulmonary tissue slices under light microscope;in the three Lianhuaqingwen groups, we found the exudation of inflammatory cells in alveolar space reduced with the increase of the dosage of Lianhuaqingwen. Sodium chloride group and control group were not significantly different in the levels of IL-1,IL-6 and TNF-α in BALF and serum(P>0. 05);PM2.5 group was higher than sodium chloride group in the level of IL-1 in BALF,and PM2. 5 group was higher than control group and sodium chloride group in the level of IL-1 in serum and the levels of IL-6 and TNF-αin BALF and serum(P<0. 05);high-dose group was lower than control group,sodium chloride group,PM2. 5 group,low-dose group and medium group in the level of IL-1 in BALF,lower than control group,sodium chloride group,PM2. 5 group and low-dose group in the level of IL-6 in BALF and serum,lower than PM2. 5 group,low-dose group and medium-dose group in the level of TNF-α in BALF and the level of IL-1 in serum, and lower than PM2. 5 group and low -dose group in the level of TNF -α in serum ( P <0. 05 ). Conclusion The acute exposure to PM2. 5 can induce pulmonary inflammatory injury in rats, Lianhuaqingwen could produce antagonistic effects to the inflammatory injury.