目的 探討漢防己甲素對脂多糖誘導(dǎo)的小鼠急性肺損傷的作用及機制。方法 將ICR小鼠根據(jù)體質(zhì)量隨機分為對照組、模型組、地塞米松組、漢防己甲素（19.5、39.0、78.0 mg/kg）組。給藥干預(yù)3 d后除對照組小鼠鼻腔滴注無菌生理鹽水外，其余各組均鼻腔滴注脂多糖生理鹽水溶液構(gòu)建急性肺損傷模型，24 h后進行對小鼠進行肺部影像學(xué)檢查；并測定小鼠肺濕質(zhì)量（W）與干質(zhì)量（D）比值；利用蘇木素–伊紅（HE）染色觀察肺組織病理形態(tài)學(xué)變化；采用酶聯(lián)接免疫吸附測定（ELISA）檢測支氣管肺泡灌洗液（BALF）中白細胞介素-1β(IL-1β)、白細胞介素-6(IL-6)和腫瘤壞死因子-α(TNF-α)含量；選用可見分光光度法檢測肺組織中髓過氧化物酶（MPO）活性；Western blotting驗證小鼠肺組織TNF-α受體1(TNFR1)與受體相互作用蛋白激酶1(RIPK1)蛋白表達水平；最后選擇免疫熒光法檢測小鼠肺組織TNFR1相對表達水平。結(jié)果 與模型組相比，漢防己甲素組小鼠肺部影像與組織病理改變減輕；漢防己甲素39.0、78.0 mg/kg組均可顯著降低W/D值（P<0.01、0.001）；并顯著降低IL-1β、IL-6、TNF-α含量（P<0.05、0.01、0.001）；且漢防己甲素78.0 mg/kg組可顯著下調(diào)MPO的活性（P<0.05）;Western blotting結(jié)果顯示，漢防己甲素39.0 mg/kg組可顯著下調(diào)RIPK1蛋白相對表達量，漢防己甲素78.0 mg/kg組可顯著下調(diào)TNFR、TNF-α的蛋白相對表達量（P<0.05）；免疫熒光結(jié)果也進一步驗證漢防己甲素組可下調(diào)TNFR的熒光表達。結(jié)論 漢防己甲素能夠減輕脂多糖誘導(dǎo)的小鼠急性肺損傷，其作用機制可能與TNF信號通路有關(guān)。

Objective To discuss the effect and mechanism of tetrandrine in mice model of acute lung injury induced by lipopolysaccharide. Methods ICR mice were randomly divided into control group, model group, dexamethasone group, tetrandrine（19.5, 39.0, and 78.0 mg/kg） group. After 3 days of drug intervention, except for the control group of mice, which received intranasal infusion of sterile physiological saline, all other groups received intranasal infusion of lipopolysaccharide physiological saline solution to construct acute lung injury model. After 24 hours, radiographic examination was performed on mice, and then mice were sacrificed to measure the ratio of wet weight（W） to dry weight（D） of the lungs, observe the pathological changes of lung tissue by HE staining.IL-1β, IL-6, and TNF-α levels in BALF were detected by ELISA. MPO activity in lung tissue was detected by visible spectrophotometry method, and Western blotting was used to verify the protein expression levels of TNFR1 and RIPK1 in mice lung tissue. Finally, immunofluorescence was used to detect the relative expression level of TNFR1 in mouse lung tissue. Results Compared with the model group, mice in the tetrandrine group had alleviated tissue pathology changes, and both tetrandrine 39.0 and 78.0 mg/kg groups could significantly reduce the W/D value（P＜0.01, 0.001）, significantly reduce IL-1β, IL-6, and TNF-α content（P＜0.05, 0.01, and 0.001）. And significantly reduce MPO activity in the tetrandrine 78.0 mg/kg group（P＜0.05）. Western blotting results showed that tetrandrine 39.0 mg/kg group could significantly downregulate RIPK1 protein expression, while the tetrandrine 78.0 mg/kg group could significantly downregulate TNFR and TNF-α protein expression（P＜0.05）. Immunofluorescence results also further verified that tetrandrine group can down-regulate the fluorescence expression of TNFR. Conclusion Tetrandrine can alleviate lipopolysaccharide induced acute lung injury in mice, and its mechanism of action may be related to the TNF signaling pathway.

R285.5

國家自然科學(xué)基金青年基金項目(82204740); 中國中醫(yī)科學(xué)院優(yōu)秀青年科技人才培養(yǎng)專項(ZZ16-YQ-027)