目的 基于網(wǎng)絡(luò)藥理學(xué)和動(dòng)物實(shí)驗(yàn)探究升降散治療急性肺損傷（ALI）的作用機(jī)制。方法 在中藥系統(tǒng)藥理學(xué)分析平臺(tái)數(shù)據(jù)庫（TCMSP）和Swiss Target Prediction數(shù)據(jù)庫中檢索升降散的成分及靶點(diǎn)，在基因數(shù)據(jù)庫（Gene Cards）中檢索ALI的疾病靶點(diǎn)。將藥物與疾病交集靶點(diǎn)上傳至STRING數(shù)據(jù)平臺(tái)進(jìn)行蛋白質(zhì)-蛋白質(zhì)相互作用（PPI）分析，運(yùn)用DAVID網(wǎng)站進(jìn)行基因本體（GO）富集分析和京都基因與基因組百科全書（KEGG）通路分析。體內(nèi)實(shí)驗(yàn)，除對(duì)照組外，升降散各給藥組分別ig給藥20 mL·kg^-1藥液（散劑、水煎液高、低劑量分別為175、350 mg·kg^-1），每天早晚各1次，共連續(xù)給藥14 d；地塞米松組第12天開始ip地塞米松（5 mg·kg^-1），連續(xù)給藥3 d；除對(duì)照組外，其余各組第15天分別向氣管內(nèi)滴注1 mg·mL^-1脂多糖（5 mg·kg^-1）進(jìn)行造模，觀察肺組織病理損傷情況，ELISA法檢測白細(xì)胞介素-6（IL-6）、白細(xì)胞介素-1β（IL-1β）、腫瘤壞死因子-α（TNF-α）水平，實(shí)時(shí)熒光半定量聚合酶鏈?zhǔn)椒磻?yīng)（qRT-PCR）法檢測Toll樣受體4 （TLR4）、磷酸化-絲裂原活化蛋白激酶p38 （p38-MAPK）、細(xì)胞外調(diào)節(jié)蛋白激酶（ERK）、c-Jun氨基末端激酶（JNK-1） mRNA表達(dá)量，Western blotting法檢測核因子κB（NF-κB）和人核因子κB抑制蛋白α（IκBα）表達(dá)量。結(jié)果 網(wǎng)絡(luò)藥理學(xué)預(yù)測出升降散活性成分32個(gè)，關(guān)鍵靶點(diǎn)292個(gè)，ALI疾病靶點(diǎn)有1 454個(gè)，兩者交集靶點(diǎn)95個(gè)；升降散治療ALI的關(guān)鍵靶點(diǎn)主要參與NF-κB、MAPK等信號(hào)通路。通過ALI小鼠模型實(shí)驗(yàn)結(jié)果顯示，升降散能夠顯著減輕ALI小鼠肺部組織炎癥，改善肺泡間隔及肺泡壁增厚程度；降低血清中炎癥因子IL-6、IL-1β、TNF-α的水平（P＜0.05、0.01）；下調(diào)TLR4、p38-MAPK、ERK、JNK-1 mRNA的表達(dá)量（P＜0.05、0.01）；Western blotting結(jié)果表明升降散可通過抑制NF-κB蛋白磷酸化水平（P＜0.01），激活I(lǐng)κBα蛋白（P＜0.01），緩解ALI。結(jié)論 升降散能夠基于多成分、多靶點(diǎn)、多通路的特點(diǎn)相互協(xié)同治療ALI，其機(jī)制可能與NF-κB、MAPK等信號(hào)通路有關(guān)，且散劑治療ALI的作用優(yōu)于水煎液。

Objective To explore the mechanism of Shengjiang Powde in the treatment of acute lung injury (ALI) based on network pharmacology and animal experiments. Methods The components and targets of Shengjiang Powder were searched in Traditional Chinese Medicine Systems Pharmacology Analysis Platform Database (TCMSP) and Swiss Target Prediction Database, and the disease targets of ALI were searched in Gene Cards Database. The drug-disease intersection targets were uploaded to the STRING data platform for protein interaction analysis. Gene ontology (GO) enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis were performed using the DAVID website. In order to further explore the effect of Shengjiang Powder on ALI, a mouse ALI model was established to observe the pathological damage of lung tissue. In addition to the control group, each administration group of Shengjiang Powder was ig administered with 20 mL·kg^?1 drug solution (the doses of powder, water decoction and its high and low dose groups were 175 and 350 mg·kg^?1, respectively, once a day in the morning and evening, for a total of 14 days). On the 12 th day, dexamethasone (5 mg·kg^?1) was injected intraperitoneally for three days. On the 15th day, 1 mg·mL^?1 lipopolysaccharide (5 mg·kg^?1) was instilled into the trachea for modeling. The levels of interleukin 6 (IL-6),interleukin 1β (IL-1β) and tumor necrosis factor-α (TNF-α) were detected by ELISA. The mRNA expressions of Toll-like receptor 4 (TLR4), phosphorylated-mitogen-activated protein kinase p38 (p38-MAPK), extracellular regulated protein kinases (ERK), c-Jun Nterminal kinase (JNK-1) were detected by RT-PCR. Protein expressions of nuclear factor kappa B (NF-κB) and human nuclear factor κB inhibitory protein α (IκBα) were detected by Western blotting. Results It was predicted that there were 32 active components, 292 key targets, 1 454 ALI disease targets and 95 intersection targets of Shengjiang Powder and ALI. The key targets of Shengjiang Powder in the treatment of ALI were mainly involved in NF-κB, MAPK and other signaling pathways. The results of ALI mouse model showed that Shengjiang Powder could significantly reduce the inflammation of lung tissue in ALI mice and improve the thickening of alveolar septum and alveolar wall. Shengjiang Powder could reduce the content of inflammatory factors IL-6, IL-1β and TNF-α in serum. The expression of TLR4, p38-MAPK, ERK and JNK-1 mRNA was down-regulated. Western blotting results showed that Shengjiang Powder could activate IκBα protein and alleviate ALI by inhibiting the phosphorylation level of NF- κB protein. Conclusion Shengjiang Powder can synergistically treat ALI based on the characteristics of multi-component, multi-target and multi-pathway. The mechanism may be related to NF-κB, MAPK and other signaling pathways, and the effect of Shengjiang Powder on ALI is better than that of water extract.

R285.5

中醫(yī)藥事業(yè)傳承發(fā)展資金支持項(xiàng)目（吉中醫(yī)藥發(fā)［2021］11號(hào)）