目的 采用網(wǎng)絡(luò)藥理學(xué)方法探討白花蛇舌草治療分化良好的胰腺神經(jīng)內(nèi)分泌腫瘤（pNET）的作用機(jī)制，并通過體外細(xì)胞實(shí)驗(yàn)進(jìn)行驗(yàn)證。方法 應(yīng)用中藥系統(tǒng)藥理學(xué)數(shù)據(jù)庫(kù)與分析平臺(tái)（TCMSP）獲取白花蛇舌草的活性成分和作用靶點(diǎn)，通過GEO、GeneCards、OMIM、PharmGkb、Drugbank數(shù)據(jù)庫(kù)獲取pNET的疾病相關(guān)靶點(diǎn)，通過R軟件對(duì)共同靶點(diǎn)進(jìn)行基因本體（GO）和京都基因和基因組百科全書（KEGG）通路富集分析。以CCK-8法和Western blotting體外實(shí)驗(yàn)觀察白花蛇舌草提取物對(duì)人胰腺神經(jīng)內(nèi)分泌腫瘤BON-1、QGP-1細(xì)胞的增殖抑制作用及對(duì)細(xì)胞凋亡相關(guān)蛋白Bcl-2表達(dá)的影響。結(jié)果 篩選得到白花蛇舌草活性成分7個(gè)，成分相關(guān)靶點(diǎn)158個(gè)；獲得pNET疾病相關(guān)靶點(diǎn)2 870個(gè)，成分與疾病共同相關(guān)靶點(diǎn)119個(gè)。共同靶點(diǎn)的GO功能富集分析共得到2 099個(gè)條目，KEGG富集分析共得到162條信號(hào)通路，包括細(xì)胞凋亡、PI3K/Akt、P53等信號(hào)通路。不同質(zhì)量濃度［0（對(duì)照）、1.0、1.5、2.0、2.5、3.0、3.5、4.0、4.5 mg·mL^-1］白花蛇舌草提取物干預(yù)BON-1、QGP-1細(xì)胞48 h后，細(xì)胞增殖明顯受到抑制，且抑制作用具有明顯的濃度相關(guān)性。與對(duì)照組比較，隨著白花蛇舌草提取物質(zhì)量濃度增加，BON-1、QGP-1細(xì)胞的增殖受到明顯抑制，漂浮死亡細(xì)胞數(shù)增加；與對(duì)照組比較，高質(zhì)量濃度白花蛇舌草提取物可顯著降低BON-1、QGP-1細(xì)胞Bcl-2蛋白表達(dá)（P < 0.05）。結(jié)論 白花蛇舌草提取物能夠有效抑制人胰腺神經(jīng)內(nèi)分泌腫瘤BON-1、QGP-1細(xì)胞增殖，通過抑制Bcl-2蛋白表達(dá)促進(jìn)細(xì)胞凋亡；白花蛇舌草能夠通過多成分、多靶點(diǎn)、多通路的途徑治療胰腺神經(jīng)內(nèi)分泌腫瘤。

Objective To investigate the mechanism of Hedyotis diffusa in treatment of well differentiated pancreatic neuroendocrine tumor (pNET) via network pharmacology and in vitro cytologic experiments. Methods The active constituents of H. diffusa and drug targets were obtained from the Traditional Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP), and the disease-related targets of pNET were retrieved from GEO, GeneCards, OMIM, PharmGkb, and Drugbank databases. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis for common targets of drugs and disease was performed using the R software. CCK-8 assay were used to verify the inhibiton of H. diffusa extract on proliferation of human pancreatic neuroendocrine tumor of BON-1 and QGP-1 cells and apoptosis related proteins of the influence of the Bcl-2 expression.. Western blotting was used to observe effect of H. diffusa extract on apoptosis of BON-1 and QGP-1 cells. Results A total of 158 drug targets of seven active ingredients of H. diffusa, and 2 870 disease-related targets orelated to pNET were obtained resulting in 119 common targets. A total of 2 099 items were obtained by GO functional enrichment analysis. A total of 162 signaling pathways were obtained by KEGG enrichment analysis, including apoptosis, PI3K-Akt, and P53 signaling pathways, etc (P < 0.05). CCK-8 assay showed that the extract of H. diffusa had a concentration-dependent inhibitory effect on proliferation of BON-1 and QGP-1 cells. Western blotting showed that the protein expression of Bcl-2 in the high-concentration extract of H. diffusa group was significantly decreased compared with the control group (P < 0.05). Conclusion The extract of H. diffusa can effectively inhibit the proliferation of human pancreatic neuroendocrine tumors of BON-1 and QGP-1 cells and promote apoptosis by inhibiting the expression of Bcl-2 protein. H. diffusa can treat pancreatic neuroendocrine tumors through multi-component, multi-target and multi-pathway approaches.

R285.5

國(guó)家重點(diǎn)研發(fā)計(jì)劃項(xiàng)目（2019YFB1309704）