目的 探討歐前胡素對人胃癌細胞(MKN-45)和人結腸癌細胞(HT-29)中煙酰胺腺嘌呤二核苷酸磷酸氧化酶(Noxs)-活性氧(ROS)氧化應激通路的作用及對腫瘤細胞增殖活性的影響。方法 MKN-45和HT-29細胞經(jīng)過不同濃度(3、10、30、100 μmol/L)的歐前胡素及陽性對照二苯基氯化碘鹽(DPI)處理后,使用CCK8法檢測細胞增殖活性,使用DHE熒光探針檢測細胞內ROS水平,使用Western blotting檢測細胞內Nox1、Nox2、Nox3、Nox4、Nox5家族蛋白表達。結果 CCK8法結果顯示,歐前胡素可濃度相關性地抑制MKN-45和HT-29細胞增殖活性;DHE熒光探針法結果顯示,歐前胡素干預可使MKN-45和HT-29細胞熒光強度明顯減弱,ROS水平降低;Western blotting結果顯示,給予歐前胡素可使MKN-45細胞Nox1、Nox2、Nox3、Nox4、Nox5蛋白表達水平顯著降低(P<0.05、0.01、0.001),使HT-29細胞Nox1、Nox2蛋白表達水平顯著降低(P<0.05、0.01)。結論 歐前胡素可通過抑制MKN-45和HT-29細胞中Noxs-ROS通路,誘導細胞增殖活性下降,發(fā)揮抗腫瘤作用。

Objective To explore the effect of imperatorin on the Noxs-ROS oxidative stress pathway and proliferative activity in human gastric cancer cells (MKN-45) and human colon cancer cells cells (HT-29). Methods MKN-45 and HT-29 cells were administered with different concentrations (3, 10, 30, and 100 μmol/L) of imperatorin and positive control diphenyl iodide salt chloride (DPI), CCK8 method was used to detectcell proliferation activity, intracellular ROS level was detected by DHE fluorescent probe, and Western blotting was used to detect the protein expressions of Nox1, Nox2, Nox3, Nox4, and Nox5. Results CCK8 assay showed that imperatorin could concentration-dependently inhibited the proliferation activity of MKN-45 and HT-29 cells. The results of DHE fluorescent probe assay showed that imperatorin intervention could significantly reduced the fluorescence intensity and ROS level of MKN-45 and HT-29 cells. Western blotting results showed that administration of imperatorin could reduced the protein expression level of Nox1, Nox2, Nox3, Nox4, and Nox5 in MKN-45, and Nox1 and Nox2 in HT-29 cells. Conclusion Imperatorincan inhibits the proliferative activity of MKN-45 and HT-29 cells by down-regulating of Noxs-ROS pathway, and exhibits anti-tumor effects.

R285.5

國家自然科學基金資助面上項目(82172418)