目的 采用網(wǎng)絡藥理學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）的作用機制，并通過體外細胞實驗進行驗證。方法 應用中藥系統(tǒng)藥理學數(shù)據(jù)庫與分析平臺（TCMSP）獲取白花蛇舌草的活性成分和作用靶點，通過GEO、GeneCards、OMIM、PharmGkb、Drugbank數(shù)據(jù)庫獲取pNET的疾病相關靶點，通過R軟件對共同靶點進行基因本體（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體外實驗觀察白花蛇舌草提取物對人胰腺神經(jīng)內(nèi)分泌腫瘤BON-1、QGP-1細胞的增殖抑制作用及對細胞凋亡相關蛋白Bcl-2表達的影響。結果 篩選得到白花蛇舌草活性成分7個，成分相關靶點158個；獲得pNET疾病相關靶點2 870個，成分與疾病共同相關靶點119個。共同靶點的GO功能富集分析共得到2 099個條目，KEGG富集分析共得到162條信號通路，包括細胞凋亡、PI3K/Akt、P53等信號通路。不同質(zhì)量濃度［0（對照）、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BON-1、QGP-1細胞48 h后，細胞增殖明顯受到抑制，且抑制作用具有明顯的濃度相關性。與對照組比較，隨著白花蛇舌草提取物質(zhì)量濃度增加，BON-1、QGP-1細胞的增殖受到明顯抑制，漂浮死亡細胞數(shù)增加；與對照組比較，高質(zhì)量濃度白花蛇舌草提取物可顯著降低BON-1、QGP-1細胞Bcl-2蛋白表達（P < 0.05）。結論 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細胞增殖，通過抑制Bcl-2蛋白表達促進細胞凋亡；白花蛇舌草能夠通過多成分、多靶點、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

國家重點研發(fā)計劃項目（2019YFB1309704）