摘要
目的观察木姜子和忍冬藤配伍醇提物对哮喘细胞外信号调节激酶1/2(ERK1/2)-白细胞介素-17(IL-17)通路及细胞增殖凋亡的影响。方法用卵清蛋白诱导哮喘大鼠获得的哮喘平滑肌细胞模型,将模型细胞分为模型组和低、中、高剂量实验组,另取正常细胞作为空白组。空白组和模型组均置于含10%胎牛血清(FBS)的DMEM/F12培养基进行培养;低、中、高剂量实验组分别置于含5.65,11.30和22.60μg·mL-1木姜子和忍冬藤配伍醇提物及10%FBS的DMEM/F12培养基进行培养。用噻唑蓝法检测增殖抑制率,用TUNEL法观察细胞凋亡,用酶联免疫吸附实验法检测IL-17和转录激活因子6(STAT-6)水平,用Western blot法检测凋亡相关因子(FAS)、ERK1/2和抗凋亡蛋白(Bcl-2)相对表达量。结果低、中、高剂量实验组的增殖抑制率分别为(0.42±0.03),(0.64±0.06)和(0.81±0.08),凋亡指数分别为(0.22±0.04),(0.27±0.05)和(0.35±0.06)。低、中、高剂量实验组和模型组、空白组的FAS蛋白相对表达量分别为(0.21±0.04),(0.64±0.05),(0.82±0.07),(0.12±0.02)和(0.72±0.06),ERK1/2蛋白相对表达量分别为(0.38±0.06),(0.33±0.05),(0.18±0.02),(0.87±0.09)和(0.32±0.05),Bcl-2蛋白相对表达量分别为(0.32±0.06),(0.28±0.05),(0.22±0.04),(0.89±0.09)和(0.26±0.06),IL-17分别为(121.02±26.65),(86.35±24.31),(61.55±22.01),(286.22±30.21)和(53.21±21.26)pg·mL-1,STAT-6分别为(189.36±26.69),(98.34±19.89),(56.39±15.61),(346.11±35.02)和(39.25±11.22)pg·mL-1。低、中、高剂量实验组的上述指标与模型组相比较,差异均有统计学意义(均P<0.05)。结论木姜子和忍冬藤配伍醇提物可抑制气道平滑肌细胞增殖,诱导气道平滑肌细胞凋亡,其机制可能与抑制ERK1/2-IL-17通路相关分子,上调凋亡相关因子FAS蛋白表达、抑制抗凋亡蛋白Bcl-2表达相关。
Objective To observe the effects of Litsea combined with Lonicera japonica Thunb on extracellular signal-regulated kinases 1 and 2(ERK1/2)-interleukin 17(IL-17) pathway, cell proliferation and apoptosis in asthma.Methods Asthma airway smooth muscle cells(ASMCs) were obtained from the asthma rat model that was induced by injection of ovalbumin and then Asthma ASMCs were incubated and divided into model,experimental-L,experimental-M and experimental-H groups,besides,normal ASMCs as blank group.The blank and model groups were cultured by 10% FBS + DMEM/F12 while the experimental-L,-M and-H groups respectively were cultured by 5.65,11.30 and 22.60 μg · mL-1(Litsea combined with Lonicera japonica) + 10% FBS + DMEM/F12.The cell proliferation inhibition rate was examined by methyl thiazolyl tetrazolium assay.The cell apoptosis was determined by TUNEL method.The levels of signal transducers and activators of transduction 6(STAT-6) and IL-17 were detected by enzyme linked immunosorbent assay.The relative protein expressions of apoptosis-related factor(FAS),ERK1/2 and B-cell lymphoma-2(Bcl-2) were detected by Western blot.Results The cell proliferation inhibition rates in the experimental-L,-M and-H groups were(0.42±0.03),(0.64±0.06) and(0.81±0.08),the apoptosis indexes were(0.22±0.04),(0.27±0.05) and(0.35±0.06).In the experimental-L,experimental-M,experimental-H, model and blank groups, the relative protein expression of FAS were(0.21±0.04),(0.64±0.05),(0.82±0.07),(0.12±0.02) and(0.72±0.06),the relative protein expression of ERK1/2 were(0.38±0.06),(0.33±0.05),(0.18±0.02),(0.87±0.09) and(0.32±0.05),the relative protein expression of Bcl-2 were(0.32±0.06),(0.28±0.05),(0.22±0.04),(0.89±0.09) and(0.26±0.06),the levels of IL-17 were(121.02±26.65),(86.35±24.31),(61.55±22.01),(286.22±30.21) and(53.21±21.26)pg·mL-1, the levels of STAT-6 were(189.36±26.69),(98.34±19.89),(56.39±15.61),(346.11±35.02) and(39.25±11.22) pg ·mL-1.There were significant differences in the above-mentioned indexes between the model group and the experimental-L,-M and-H groups(all P < 0.05).Conclusion Litsea combined with Lonicera japonica was suggested to inhibit proliferation of airway smooth muscle cell and induce apoptosis.The possible mechanism might be related to the inhibition of ERK1/2-IL-17 pathway,the upregulation of the protein expression of FAS,and the inhibition of Bcl-2 expression.
作者
黄小珊
蓝凰齐
唐汉庆
李克明
梁凌玲
赵秋华
HUANG Xiao-shan;LAN Huang-qi;TANG Han-qing;LI Ke-ming;LIANG Ling-ling;ZHAO Qiu-hua(.School of Basic Medicine,Youjiartg Medical University for Nationalities,Baise 533000,Guangxi Zhuang Autonomous Region,China;School of Basic Medicine,Guangxi University of Chinese Medicine,Naming 530001,Guangxi Zhuang Autonomous Region,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2020年第7期775-778,共4页
The Chinese Journal of Clinical Pharmacology
基金
国家自然科学基金资助项目(81760759)。
关键词
哮喘
气道平滑肌细胞
细胞外信号调节激酶1/2
凋亡
asthma
airway smooth muscle cell
extracellular signal-regulated kinases 1 and 2
apoptosis
