脂肪乳通过LPS/TLR4通路对急性有机磷中毒肺损伤的治疗作用研究  

Therapeutical effect of lipid emulsion in lung damage induced by acute organophosphorus poisoning through LPS/TLR4 pathway

作　　者：李刚[1] 韩楠楠[1] 张可[1] 李恒杰[1] LI Gang;HAN Nannan;ZHANG Ke;LI Hengjie(Department of Emergency Medicine,Zhejiang Provincial People′s Hospital/Affiliated People′s Hospital of Hangzhou Medical College,Hangzhou,Zhejiang 310014,China)

机构地区：[1]浙江省人民医院/杭州医学院附属人民医院急诊医学科,杭州310014

出　　处：《重庆医学》2022年第9期1448-1452,共5页Chongqing medicine

基　　金：浙江省医药卫生科技计划项目(2020KY441)。

摘　　要：目的探讨脂肪乳对急性有机磷中毒所致肺损伤的治疗效果及脂多糖(LPS)/Toll样受体4(TLR4)通路在治疗过程中发挥的作用。方法将50只大鼠随机分为A组(盐水对照组)、B组(脂肪乳对照组)、C组(标准治疗组)、D组(脂肪乳治疗组)、E组(LPS/TLR4拮抗剂组),给予90%敌敌畏(7 mg/kg)腹腔注射染毒。A组染毒后尾静脉注射生理盐水(5 mL/kg);B组染毒后尾静脉注射20%脂肪乳(5 mL/kg);C组染毒后予阿托品(10 mg/kg)和氯解磷定(40 mg/kg)肌肉注射;D组在C组的基础上尾静脉注射20%脂肪乳(5 mL/kg);E组在D组的基础上尾静脉注射LPS/TLR4拮抗剂(0.5 mg/kg)。染毒后4 h使用毛细导管眶下采血1 mL行全血胆碱酯酶检查;染毒后24 h内观察大鼠流涎、肌肉震颤等中毒表现和存活情况;染毒后24 h取肺组织,HE染色电镜下观察肺组织病理结构,利用荧光定量PCR技术检测肺组织中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、核因子-κB(NF-κB)mRNA相对表达量,流式细胞术检测肺细胞悬液中LPS/TLR4表达。结果敌敌畏染毒后,A组中毒症状出现较早,并且实验大鼠均出现了流涎、肌束震颤、躁动等中毒症状。染毒后24 h,A组有3只大鼠存活,B组有4只大鼠存活,C组有5只大鼠存活,D组有7只大鼠存活,而E组存活9只,仅2只大鼠出现流涎。E组存活率、胆碱酯酶活性最高,而流涎率、肌束震颤评分最低,与其他4组比较差异有统计学意义(P<0.05)。HE染色电镜下肺组织病理结果显示,D组较A、B、C 3组炎症细胞浸润、红细胞渗出明显减轻,E组肺损伤最轻。染毒后24 h肺组织IL-6、NF-κB、TNF-α相对表达量,B组低于A组,D组低于A、B、C 3组,E组低于其他4组,差异有统计学意义(P<0.01)。染毒后24 h肺细胞悬液中LPS/TLR4表达,B组低于A组,D组低于A、B、C 3组,E组低于其他4组,差异有统计学意义(P<0.01)。结论脂肪乳可以减轻急性有机磷中毒所致肺损伤,LPS/TLR4通路在治疗过程中发Objective To explore the therapeutical effect of lipid emulsion in lung damage induced by acute organophosphorus poisoning and its effect of LPS/TLR4 pathway in the treatment process.Methods Fifty Sprague-Dawley(SD)rats were random divided into the five groups:A,B,C,D and E.90%dichlorvos(DDVP,7 mg/kg)was intraperitoneally injected for poison contamination.The group A was given normal saline(5 mL/kg)by tail venous injection after poison contamination;20%lipid emulsion(5 mL/kg)was given by tail intravenous injection in the group B;atropine(5 mL/kg)and pralidoxime chloride(40 mL/kg)were given by intramuscular injection in the group C;on the basis of standard treatment group,the combined use of 20%lipid emulsion(5 mL/kg)was given by tail vein injection in the group D;on the lipid emulsion treatment,the LPS/TLR4 antagonist(0.5 mg/kg)was given by tail vein injection in the group E.1 mLblood was collected under the orbit by capillary tube for detecting the whole blood cholinesterase(CHE)at 4 hours after DDVP poisoning.The poisoning manifestations such as salivation and muscle tremor,and survival situation were observed within 24 h after poisoning.The lung tissues were taken after 24 h of poisoning for observing the lung tissues pathological structure by HE staining.The fluorogenic quantitative PCR was applied to detect the relative expression levels of IL-6,TNF-α,NF-κB in lung tissue.The flow cytometry was applied to detect the expression level of LPS/TLR4 in lung cell suspension.Results After DDVP poisoning,the poisoning symptoms in the group A appeared earlier,such as muscular tremor,salivation and restlessness.After DDVP poisoning,only 3 mice in the group A survived,which had 4 mice in the group B,5 mice in the group D,7 mice in the group D,and 9 mice in the group E,only 2 mice had salvation.Compared with the group A,B,C and D,the survival rate and cholinesterase activity in the group E were the highest,while the salivation rate and the intensity of muscular tremor were the lowest,and the differences were statistic

关 键 词：脂肪乳 LPS/TLR4 急性有机磷中毒 肺损伤 

分 类 号：R595.4[医药卫生—内科学]