漢黃芩素通過激活活性氧簇介導(dǎo)的p38MAPK信號通路誘導(dǎo)類風(fēng)濕關(guān)節(jié)炎成纖維樣滑膜細(xì)胞凋亡
發(fā)布時間：2018-09-15 19:19
【摘要】：目的探討漢黃芩素對類風(fēng)濕關(guān)節(jié)炎成纖維樣滑膜細(xì)胞(rheumatoid arthritis fibroblast-like synoviocytes,RA-FLS)凋亡的影響以及作用機(jī)制。方法類風(fēng)濕關(guān)節(jié)炎患者關(guān)節(jié)滑液經(jīng)原代培養(yǎng),應(yīng)用3~5代傳代的成纖維樣滑膜細(xì)胞。按照不同方法處理分為6組:空白對照組、低劑量漢黃芩素組(終濃度為20μg/mL)、中劑量漢黃芩素組(終濃度為50μg/mL)、高劑量漢黃芩素組(終濃度為100μg/mL)、100μg/mL漢黃芩素+NAC組、100μg/mL漢黃芩素+SB203580組。MTT法檢測細(xì)胞增殖情況,流式細(xì)胞儀檢測細(xì)胞凋亡率,羅丹明123染色熒光顯微鏡照相法檢測線粒體膜電位(mitochondrial membrane potential,MMP),DCFH-DA染色熒光顯微鏡照相法檢測細(xì)胞內(nèi)活性氧簇(reactive oxygen species,ROS)水平,Western blot法檢測p38MAPK通路相關(guān)蛋白的表達(dá)。結(jié)果與正常對照組相比,各劑量漢黃芩素均能夠抑制RA-FLS細(xì)胞的增殖,增加其凋亡,降低MMP以及增加ROS水平,激活p38MAPK信號通路,并且呈現(xiàn)劑量依賴性關(guān)系。ROS清除劑NAC和p38抑制劑SB203580可以明顯降低漢黃芩素誘導(dǎo)的RA-FLS細(xì)胞的凋亡,并且5 mmol/L NAC下調(diào)漢黃芩素引起的p38MAPK磷酸化。結(jié)論漢黃芩素能夠誘導(dǎo)RA-FLS細(xì)胞凋亡,ROS/p38MAPK信號通路參與了其凋亡的過程。
[Abstract]:Objective to investigate the effect of baicalin on apoptosis of fibroid synovial cells (rheumatoid arthritis fibroblast-like synoviocytes,RA-FLS) in rheumatoid arthritis (RA) and its mechanism. Methods the synovial fluid of patients with rheumatoid arthritis was cultured in primary culture. According to different methods, they were divided into 6 groups: blank control group, The cell proliferation was detected with low dose of baicalin (20 渭 g/mL), middle dose (50 渭 g/mL) and high dose (final concentration of 100 渭 g/mL) of 100 渭 g/mL and 100 渭 g/mL SB203580. The apoptosis rate was detected by flow cytometry. Detection of mitochondrial membrane potential (mitochondrial membrane potential,MMP) by Rhodamine 123 staining fluorescence microscope) DCFH-DA staining fluorescence microscopy was used to detect the level of reactive oxygen species (reactive oxygen species,ROS) in cells. Western blot was used to detect the expression of p38MAPK pathway related proteins. Results compared with the control group, baicalin could inhibit the proliferation of RA-FLS cells, increase its apoptosis, decrease the level of MMP and increase the level of ROS, and activate the p38MAPK signaling pathway. In a dose-dependent manner, Ros scavenger NAC and p38 inhibitor SB203580 could significantly reduce the apoptosis of RA-FLS cells induced by baicalin, and 5 mmol/L NAC down-regulated p38MAPK phosphorylation induced by baicalin. Conclusion: baicalin can induce apoptosis of RA-FLS cells via Ros / p38 MAPK signaling pathway.
【作者單位】： 河南省中醫(yī)院風(fēng)濕病科;河南中醫(yī)藥大學(xué)第一附屬醫(yī)院風(fēng)濕病科;
【分類號】：R259
[Abstract]:Objective to investigate the effect of baicalin on apoptosis of fibroid synovial cells (rheumatoid arthritis fibroblast-like synoviocytes,RA-FLS) in rheumatoid arthritis (RA) and its mechanism. Methods the synovial fluid of patients with rheumatoid arthritis was cultured in primary culture. According to different methods, they were divided into 6 groups: blank control group, The cell proliferation was detected with low dose of baicalin (20 渭 g/mL), middle dose (50 渭 g/mL) and high dose (final concentration of 100 渭 g/mL) of 100 渭 g/mL and 100 渭 g/mL SB203580. The apoptosis rate was detected by flow cytometry. Detection of mitochondrial membrane potential (mitochondrial membrane potential,MMP) by Rhodamine 123 staining fluorescence microscope) DCFH-DA staining fluorescence microscopy was used to detect the level of reactive oxygen species (reactive oxygen species,ROS) in cells. Western blot was used to detect the expression of p38MAPK pathway related proteins. Results compared with the control group, baicalin could inhibit the proliferation of RA-FLS cells, increase its apoptosis, decrease the level of MMP and increase the level of ROS, and activate the p38MAPK signaling pathway. In a dose-dependent manner, Ros scavenger NAC and p38 inhibitor SB203580 could significantly reduce the apoptosis of RA-FLS cells induced by baicalin, and 5 mmol/L NAC down-regulated p38MAPK phosphorylation induced by baicalin. Conclusion: baicalin can induce apoptosis of RA-FLS cells via Ros / p38 MAPK signaling pathway.
【作者單位】： 河南省中醫(yī)院風(fēng)濕病科;河南中醫(yī)藥大學(xué)第一附屬醫(yī)院風(fēng)濕病科;
【分類號】：R259
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