目的 揭示白及Bletilla striata提取物抗腫瘤的有效成分和潛在分子機(jī)制。方法 首先通過生物活性追蹤方法鑒定白及中抗腫瘤活性成分。采用CCK-8法檢測化合物對膀胱癌5637細(xì)胞的毒性，進(jìn)一步篩選出最優(yōu)化合物。激光共聚焦顯微鏡觀察最優(yōu)化合物對膀胱癌細(xì)胞凋亡誘導(dǎo)。Transwell侵襲實(shí)驗(yàn)檢驗(yàn)最優(yōu)化合物對膀胱癌細(xì)胞侵襲的影響，Western blotting檢測細(xì)胞內(nèi)上皮鈣黏蛋白（epithelial cadherin，E-cadherin）、神經(jīng)鈣黏蛋白（neural-cadherin，N-cadherin）、波形蛋白（‌Vimentin）、橋粒斑蛋白（desmoplakin，DSP）、胞質(zhì)緊密粘連蛋白1（zona occludens 1，ZO-1）、細(xì)胞外調(diào)節(jié)蛋白激酶（extracellular regulated protein kinases，ERK）、磷酸化細(xì)胞外蛋白調(diào)節(jié)激酶（phospho-extracellular signal-regulated kinase，p-ERK）蛋白表達(dá)水平。結(jié)果 從白及干燥塊莖中分離鑒定出12個菲類化合物，分別為2,7-二羥基-4-甲氧基-9,10-二氫菲（1）、紅門蘭醇（2）、次苦參素（3）、2,7-二羥基-1-(對羥基芐基)-4-甲氧基-9,10-二氫菲（4）、4,7-二羥基-2-甲氧基-9,10-二氫菲（5）、2,7-二羥基-4-甲氧基菲（6）、2-羥基-4,7-二甲氧基菲（7）、8-(4-(甲酰氧基)芐基)-7-甲氧基-9,10-二氫菲-2,5-二基二甲酸酯（8）、白及聯(lián)菲A（9）、7-羥基-2-甲氧基-9,10-二氫菲-1,4-二酮（10）、7-羥基-2-甲氧基-9,10-二氫菲-1,4-二酮（11）、7-羥基-2-甲氧基-菲-1,4-二酮（12）?；衔?b>1在40 μg/mL質(zhì)量濃度下，對膀胱癌細(xì)胞5637抑制率為（53.25±1.35）%，在10、40、80 μg/mL質(zhì)量濃度下，侵襲細(xì)胞數(shù)量顯著減少，分別為（21.31±5.61）%、（54.72±0.98）%和（83.12±0.98）%。結(jié)論 化合物8、10～12為首次從該植物中分離得到?；衔?b>1可抑制ERK信號通路的激活來阻止膀胱癌5637細(xì)胞的增殖，誘導(dǎo)細(xì)胞的凋亡，并通過EMT進(jìn)程抑制人膀胱癌5637細(xì)胞的侵襲。

Objective To reveal the anti-tumor active components and molecular mechanisms of Bletilla striata. Methods The toxicity of the compound to bladder cancer cell line 5637 was detected by CCK-8 method. The apoptosis of bladder cancer cells induced by coelonin solution was observed. Transwell invasion assay was used to examine the effect of coelonin on the invasion of bladder cancer cells, and western blot assay was used to detect the expression levels of E-cadherin, N-cadherin, vimentin, desmoplakin, Zona occludens 1, extracellular regulated protein kinases (ERK), phospho-extracellular signal-regulated kinase (P-ERK) protein in cells. Results A total of 12 compounds were isolated from B. striata rhizome, respectively coelonin (1), orchinol (2), lusianthridin (3), shancidin (4), 4,7-dihydroxy-2-methoxy-9,10-dihydrophenanthrene (5), flavanthrinin (6), 2-hydroxy-4,7-dimehoxy-phenanthrene (7), 8-(4-(formyloxy)benzyl)-7-methoxy-9,10-dihydrophenanthrene-2,5-diyl diformate (8), blestriarene A (9), ephemeranthoquinone (10), ochrone A (11), densiflorol B (12). Compound 1 at 40 μg/mL showed a (53.25 ±1.35)% inhibition rate on bladder cancer cell line 5637. At different concentrations (low, medium and high), the number of invasive cells decreased significantly, which were (21.31 ±5.61)%, (54.72 ±0.98)% and (83.12 ±0.98)%, respectively. Conclusion Compounds 8, 10—12 are identified from B. striata genus for the first time. Coelonin can inhibit the activation of ERK signaling pathway to prevent the proliferation of bladder cancer 5637 cells, and inhibit the invasion of cells through EMT process. The antitumor activity of coelonin, as the main active ingredient of B. striata, is worthy of further study.

R284.1

湖南省教育廳優(yōu)秀青年項(xiàng)目（23B0751）;湖南省區(qū)域聯(lián)合基金項(xiàng)目（2024JJ7198）