目的 基于網(wǎng)絡(luò)藥理學(xué)和動(dòng)物實(shí)驗(yàn)探究六神凝膠治療痤瘡的作用機(jī)制。方法 通過網(wǎng)絡(luò)藥理學(xué)篩選六神凝膠的活性成分和作用靶點(diǎn)，采用String數(shù)據(jù)庫(kù)對(duì)活性成分靶點(diǎn)與痤瘡疾病靶點(diǎn)進(jìn)行蛋白互作網(wǎng)絡(luò)分析，并通過DAVID數(shù)據(jù)庫(kù)對(duì)六神凝膠治療痤瘡的靶點(diǎn)進(jìn)行基因本體（gene ontology，GO）功能及京都基因與基因組百科全書（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析。大鼠耳部皮內(nèi)注射痤瘡丙酸桿菌聯(lián)合外部涂抹油酸建立痤瘡模型，考察六神凝膠對(duì)大鼠耳部外觀、病理變化、耳部組織炎性因子水平的影響，采用Western blotting檢測(cè)大鼠耳部組織Toll樣受體2（Toll-like receptor 2，TLR2）/p38/核因子-κB（nuclear factor-κB，NF-κB）通路相關(guān)蛋白表達(dá)，采用免疫熒光檢測(cè)大鼠耳部組織NF-κB p65表達(dá)。結(jié)果 共獲得六神凝膠活性成分27個(gè)，藥物靶點(diǎn)831個(gè)，痤瘡靶點(diǎn)120個(gè)，活性成分與疾病的交集靶點(diǎn)41個(gè)。六神凝膠治療痤瘡的主要成分為麝香酮、膽固醇、菜油甾醇、膽酸和嗎啡等，核心靶點(diǎn)為腫瘤壞死因子（tumor necrosis factor，TNF）、白細(xì)胞介素-1β（interleukin-1β，IL-1β）、瘦素（leptin，LEP）、TLR4和CXC趨化因子配體8（CXC chemokine ligand 8，CXCL8）等，關(guān)鍵信號(hào)通路可能包括IL-17信號(hào)通路、TLR信號(hào)通路、NOD樣受體信號(hào)通路、C型凝集素受體信號(hào)通路、TNF信號(hào)通路、NF-κB信號(hào)通路等。動(dòng)物實(shí)驗(yàn)結(jié)果表明，六神凝膠能夠顯著緩解大鼠耳部痤瘡癥狀，減輕大鼠耳腫脹程度，顯著抑制耳組織中TNF-α、IL-1β、IL-6、IL-8、IL-17、IL-23和基質(zhì)金屬蛋白酶9（matrix metalloprotein 9，MMP9）水平（P＜0.001），并顯著降低痤瘡組織中TLR2、p38和p-NF-κB p65蛋白表達(dá)水平（P＜0.01、0.001）。結(jié)論 六神凝膠通過抑制TLR2/p38/NF-κB信號(hào)通路的活化，減少炎癥因子的分泌，減輕痤瘡模型大鼠炎癥反應(yīng)，從而治療痤瘡。

Objective To explore the mechanism of Liushen Gel (六神凝膠) in treatment of acne based on network pharmacology and animal experiments. Methods The active ingredients and targets of Liushen Gel were screened by network pharmacology, and the protein interaction network between the active ingredient targets and acne disease targets was analyzed by String database. Gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis of targets of Liushen Gel for acne treatment were conducted by DAVID database. The rat model of acne was established by intradermal injection of Propionibacterium acnes and external application of oleic acid. The effects of Liushen Gel on the appearance, pathological changes, and levels of inflammatory factors in ear tissue of rats were investigated. Western blotting was used to detect the expressions of Toll-like receptor 2 (TLR2)/p38/nuclear factor-κB (NF-κB) pathway related proteins in ear tissue of rats, and immunofluorescence was used to detect the expression of NF-κB p65 in ear tissue of rats. Results A total of 27 active ingredients from Liushen Gel, 831 drug targets, 120 acne targets and 41 cross targets of active ingredients and diseases were obtained. The main ingredients of Liushen Gel in the treatment of acne were muscone, cholesterol, campesterol, cholalic acid and morphine, the core targets were tumor necrosis factor (TNF), interleukin-1β (IL-1β), leptin (LEP), TLR4 and CXC chemokine ligand 8 (CXCL8), the key signal pathways may included IL-17 signaling pathway, TLR signaling pathway, NOD like receptor signaling pathway, C-type lectin receptor signaling pathway, TNF signaling pathway and NF-κB signaling pathway, etc. The animal experiment results showed that Liushen Gel could significantly alleviate the symptoms of acne in ear of rats, reduce the degree of ear swelling, significantly inhibit the levels of TNF-α, IL-1β, IL-6, IL-8, IL-17, IL-23 and matrix metalloproteinase 9 (MMP9) in ear tissue (P < 0.001), significantly reduce the expression levels of TLR2, p38 and p-NF-κB p65 proteins in acne tissue (P < 0.01, 0.001). Conclusion Liushen Gel could treat acne by inhibiting the activation of TLR2/p38/NF-κB signaling pathway, reducing the secretion of inflammatory factors and alleviating the inflammatory response of acne model rats.

R285.5

雷允上藥業(yè)集團(tuán)有限公司技術(shù)服務(wù)項(xiàng)目（XS-LS-YL-202201）；廣東省中醫(yī)藥局項(xiàng)目（20221222）；廣州市基礎(chǔ)研究計(jì)劃基礎(chǔ)與應(yīng)用基礎(chǔ)研究項(xiàng)目（202201010293）