被动吸烟合并气管注射脂多糖对大鼠肺脏和肾脏TLR信号通路的影响

Effect of Passive Smoking Combined with Tracheal Injection of Lipopolysaccharide on TLR Signaling Pathway in Rats

目的研究被动吸烟合并气管注射脂多糖(Lipopolysaccharide,LPS)对大鼠肺和肾组织的影响。方法 20只SPF级SD大鼠分为空白组和模型组。模型组大鼠每天烟熏2次,每次30 min,持续烟熏8周。模型组分别于第14、21、28、35、49天上午,大鼠腹腔注射10%的水合氯醛(3.5 mL/kg)麻醉,暴露气管,快速注入脂多糖(0.4 mg/mL),注入体积为1 mL/kg。末次烟熏后大鼠腹腔注射10%的水合氯醛(3.5 mL/kg)麻醉,取肺脏和肾脏于4%多聚甲醛溶液中固定,用于苏木精-伊红染色法(HE)及免疫组化染色。观察各组大鼠肺脏和肾脏组织的病理改变及Toll样受体4抗体(Toll-like receptor 4,TLR-4)、核因子-κB P65(nuclear factor kappa-B P65,NF-κB P65)、肿瘤坏死因子α(Tumour Necrosis Factor Alpha,TNF-α)和P物质抗体(Substance P,SP)在肺脏和肾脏组织的表达。结果HE染色结果显示模型组大鼠肺脏和肾脏组织均出现细胞排列紊乱,细胞结构异常,炎症细胞浸润等现象,经被动吸烟合并气管注射LPS对大鼠肺脏和肾脏组织均造成了不同程度的损伤。与空白组相比模型组肺脏和肾脏组织TLR-4、NF-κB P65、TNF-α、SP表达均显著增加(P<0.01或P<0.05)。肺组织TLR-4、NF-κB P65及TNF-α表达与肾脏组织TLR-4、NF-κB P65及TNF-α表达之间具有一定程度的相关性(P<0.01或P<0.05);肺组织TLR-4、NF-κB P65及SP表达与肾脏组织SP表达之间呈一定程度的相关性(P<0.01或P<0.05)。结论被动吸烟合并气管注射LPS对大鼠肺脏和肾脏组织均可产生病理损伤且二者损伤之间具有相关性,该损伤可引起TLR-4信号通路高表达。

Objective To study the effects of passive smoking combined with tracheal injection of lipopolysaccharide(LPS)on lung and kidney of rats. Methods 20 SPF SD rats were divided into blank group and model group. The rats in the model group passively smoked twice a day,30 minutes each time,lasts 8 weeks. On days 14,21,28,35,and 49,rats in the model group were anesthetized with intraperitoneal injection of 10% chloral hydrate(3.5 mL/kg),exposed the trachea,and rapidly injected lipopolysaccharide(0.4 mg/mL). After the last passive smoking,the rats were anesthetized with 10% chloral hydrate(3.5 mL/kg)intraperitoneally,the lung and kidney were fixed in 4% paraformaldehyde solution for HE staining and immunohistochemical staining. To observe the pathological changes of lung and kidney in each group and the expression of TLR-4,NF-κB P65,TNF-α and SP in lung and kidney. Results HE staining results showed that the lung and kidney of the model group had cell arrangement disorder,abnormal cell structure,and inflammatory cell infiltration. Passive smoking combined with tracheal injection of LPS damaged rat lung and kidney. Compared with the blank group,the expressions of TLR-4,NF-κB P65,and SP in the lung and kidney of the model group were significantly increased(P<0.01 or P<0.05);There was a certain degree of correlation between the expression of TLR-4,NF-κB P65,and TNF-α in lung and the expression of TLR-4,NF-κB P65,and TNF-α in kidney(P<0.01 or P<0.05);The expression of TLR-4,NF-κB P65 and SP in lung was correlated to the expression of SP in kidney with a certain extent(P<0.01 or P<0.05). Conclusion Passive smoking combined with tracheal injection of LPS can cause pathological damage to both lung and kidney of rats,and there is a correlation between lung injury and kidney injury,which can cause high expression of TLR-4 signaling pathway.