化痰降气胶囊含药血清调控Nrf2/HDAC2改善16HBE细胞糖皮质激素抵抗  

Huatanjiangqi capsule regulates Nrf2/HDAC2 and improves glucocorticoid resistance of 16HBE cells

作　　者：王梦文 郭艳 王重阳 陶福林 朱文涛 韩智莉 孙念霞 李泽庚[3] 汪电雷 WANG Mengwen;GUO Yan;WANG Chongyang;TAO Fulin;ZHU Wentao;HAN Zhili;SUN Nianxia;LI Zegeng;WANG Dianlei(School of Pharmacy,Anhui University of Chinese Medicine,Anhui 230012,Hefei,China;Department of Pharmacy,Fuyang Sixth People's Hospital,Anhui 236000,Fuyang,China;The First Afliated Hospital to Anhui University of Chinese Medicine,Anhui 230031,Hefei,China;Anhui Province Key Laboratory of Research&Development of Chinese Medicine,Anhui 230012,Hefei,China)

机构地区：[1]安徽中医药大学药学院,安徽合肥230012 [2]阜阳市第六人民医院药学部,安徽阜阳236000 [3]安徽中医药大学第一附属医院,安徽合肥230031 [4]安徽省中药研究与开发重点实验室,安徽合肥230012

出　　处：《中国临床药理学与治疗学》2021年第12期1360-1369,共10页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金资助项目(81473536);安徽省杰出青年科学基金(2108085J45)。

摘　　要：目的:探讨化痰降气胶囊含药血清(HTJQ)对香烟烟雾提取物(cigarette smoke extract, CSE)刺激下的人支气管上皮细胞(16HBE)糖皮质激素(glucocorticoid, GC)抵抗的影响。方法:16HBE细胞经HTJQ处理后,CCK-8法测定不同浓度HTJQ对16HBE细胞活力的影响。HTJQ预先处理16HBE细胞,然后依次用地塞米松(DEX)和脂多糖(LPS)培养24 h,酶联免疫吸附试验(ELISA)测定HTJQ对16HBE细胞糖皮质激素抵抗的影响。Western blot法分别测定HTJQ、萝卜硫素(SFN)和谷胱甘肽(GSH)对CSE刺激下16HBE细胞NF-E2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)、组蛋白去乙酰化酶2(HDAC2)的表达;ELISA分别测定HTJQ、SFN和GSH对16HBE细胞白细胞介素-8(IL-8)的影响。结果:HTJQ在1 h、2 h、4 h下均促进16HBE细胞增殖;ELISA和Western blot结果显示,CSE导致16HBE细胞GC抵抗(P<0.01),Nrf2、HO-1、HDAC2的表达降低(P<0.01);HTJQ显著降低IL-8,改善16HBE细胞GC敏感性(P<0.01),并上调Nrf2、HO-1、HDAC2的表达(P<0.01)。此外HTJQ显著上调16HBE细胞内GSH水平(P<0.01)。Nrf2激动剂SFN和GSH均显著改善16HBE细胞的糖皮质激素敏感性(P<0.01),上调Nrf2、HO-1、HDAC2的表达(P<0.01)。结论:HTJQ胶囊通过上调Nrf2/HDAC2蛋白表达和细胞内GSH水平,从而改善16HBE细胞GC抵抗。AIM: To explore the effect of Huatanjiangqi capsule medicated serum(HTJQ) on the resistance of human bronchial epithelial cells(16 HBE) to glucocorticoid(GC) stimulated by cigarette smoke extract(CSE). METHODS: After 16 HBE cells were treated with HTJQ, the effects of different concentrations of HTJQ on the viability of 16 HBE cells were determined by CCK-8 method. 16 HBE cells were pretreated with HTJQ, and then cultured with dexamethasone(DEX) and lipopolysaccharide(LPS) for 24 hours, the effect of HTJQ on glucocorticoid(GC) resistance of 16 HBE cells was determined by Enzyme-linked immunosorbent assay(ELISA). The effects of HTJQ, sulforaphane(SFN) and glutathione(GSH) on the expression of NF-E2-related factors 2(Nrf2), Heme oxygenase-1(HO-1) and histone deacetylase 2(HDAC2) in 16 HBE cells stimulated by CSE were measured by Western blot, and the effects of HTJQ, SFN and GSH on interleukin-8(IL-8) in 16 HBE cells were measured by ELISA. RESULTS: HTJQ promoted the proliferation of 16 HBE cells at 1 h, 2 h and 4 h, the results of ELISA and Western blot showed that CSE induced GC resistance and decreased the expression of Nrf2, HO-1 and HDAC2 in 16 HBE cells, HTJQ significantly decreased IL-8 and improved GC sensitivity of 16 HBE cells(P<0.01), and up-regulated the expression of Nrf2, HO-1 and HDAC2(P<0.01). In addition, HTJQ significantly up-regulated the level of GSH in 16 HBE cells(P<0.01). Nrf2 agonists SFN and GSH significantly improved the glucocorticoid sensitivity of 16 HBE cells(P<0.01), and up-regulated the expression of Nrf2, HO-1 and HDAC2(P<0.01). CONCLUSION: HTJQ improves the GC resistance of 16 HBE cells by up-regulating the expression of Nrf2/HDAC2 protein and the level of intracellular GSH.

关 键 词：化痰降气胶囊 慢性阻塞性肺病 糖皮质激素抵抗 NRF2 组蛋白去乙酰化酶2 谷胱甘肽 

分 类 号：R965.2[医药卫生—药理学]