摘要
目的 探讨罗红霉素（RXM）对烟草烟雾暴露下人支气管上皮细胞糖皮质激素抵抗的作用及其机制。方法 将Beas－2b细胞株分为对照组、10%烟草烟雾提取物（CSE）组、RXM＋10%CSE组。用10%CSE干预10%CSE组、10%CSE和RXM共同干预RXM＋10%CSE组、完全培养液干预对照组。酶联免疫吸附（ELISA）法检测白细胞介素8（IL－8）表达水平，并计算IL－8抑制率与地塞米松半数抑制率（IC50－Dex）；免疫荧光法和Western印迹法检测细胞组蛋白去乙酰化酶（HDAC）2蛋白相对表达量。结果 地塞米松浓度为10－9、10－8、10－7、10－6 mol/L时，RXM＋10%CSE组IL－8的抑制率[（27.55±3.81）%、（49.60±1.45）%、（55.36±3.36）%、（60.32±3.13）%]均显著低于对照组[（32.85±2.56）%、（57.12±2.81）%、（60.81±2.08）%、（67.24±3.50）%]但高于10%CSE组[（19.15±1.69）%、（37.02±2.30）%、（47.15±2.01）%、（52.09±1.57）%]（均P〈0.05）。RXM＋10%CSE组IC50－Dex[（4.94±1.62）×10－8]显著高于对照组[（1.75±0.77）×10－8]但低于10%CSE组[（2.92±0.78）×10－7]（均P〈0.01）。免疫荧光法和Western印迹法检测10%CSE组HDAC2蛋白相对表达量（0.011±0.004和0.46±0.10）均显著低于对照组（0.037±0.005和0.91±0.06），而RXM＋10%CSE组（0.025±0.005和0.77±0.09）均显著低于对照组但高于10%CSE组（均P〈0.05）。结论 RXM可通过上调HDAC2蛋白表达抑制烟草烟雾暴露诱导的人支气管上皮细胞糖皮质激素的抵抗。
Objective To explore the effects of roxithromycin （RXM） on glucocorticoid resistance of human bronchial epithelial cells exposed to smoke and its mechanism.Methods Beas-2B cells as the research object were grouped into： control group, 10%cigarette smoke extract （CSE） group, roxithromycin （RXM）＋ 10%CSE group. With 10%CSE intervention in the 10%CSE group, 10%CSE and RXM intervention in the RXM＋ 10%CSE group, complete culture solution intervention in the control group. Interleukin-8 （IL-8） levels were measured by enzyme linked immunosorbent assay （ELISA） and IL-8 inhibition rate and dexamethasone half inhibitory concentration （IC50-Dex） were calculated; the expression of histone deacetylase 2 （HDAC2） protein was detected by immunofluorescence （IF） and Western blotting （WB）.Results In response to dexamethasone at the concentration of 10-9, 10-8, 10-7 and 10-6 mol/L successively, the IL-8 inhibition rates of RXM＋ 10%CSE group [（27.55±3.81）%, （49.60±1.45）%, （55.36±3.36）%, （60.32±3.13）%, respectively] were lower than those of control group [（32.85±2.56）%, （57.12±2.81）%, （60.81±2.08）%, （67.24±3.50）%, respectively], but higher than those of 10%CSE group [（19.15±1.69）%, （37.02±2.30）%, （47.15±2.01）%, （52.09±1.57）%, respectively] （all P〈0.05）. In contrast, the IC50-Dex of RXM＋ 10%CSE group [（4.94±1.62）×10-8] was significantly higher than that of control group [（1.75±0.77）×10-8], but lower than that of 10%CSE group [（2.92±0.78）×10-7] （both P〈0.01）. The expression of HDAC2 protein of 10%CSE group （0.011±0.004 from IF and 0.46±0.10 from WB） was lower than that of control group （0.037±0.005 and 0.91±0.06, correspondingly）, while RXM＋ 10%CSE group （0.025±0.005 and 0.77±0.09, correspondingly） was lower than that of control group but higher than that of 10%CSE group （all P〈0.05）.Conclusion Roxithromycin may restrain tobacco smoke exposure-induced glucocorticoid resistance in human bronchial epithelial cells through upregulating HDAC2 expression.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2017年第28期2215-2219,共5页
National Medical Journal of China
基金
国家自然科学基金（81570027）
浙江省自然科学基金（LY16H010007）
