芍药苷调节人支气管上皮细胞系16HBE细胞表达HBD-2的分子机制研究
发布时间：2019-02-28 09:48
【摘要】：[目的]本研究旨在探讨芍药苷能否调节支气管上皮细胞系16HBE表达HBD-2及其相关的信号通路。 [方法]1.CCK8法测定不同浓度芍药苷作用下16HBE细胞活性。2.实时荧光定量RT-PCR测试芍药苷刺激16HBE细胞HBD-2mRNA的动力学表达。3.免疫印迹法(Western Blotting)测定芍药苷刺激16HBE细胞后p-ERK、ERK、p-P38MAPK、P38MAPK和p-JNK、JNK表达水平。4细胞免疫荧光法检测芍药苷刺激16HBE细胞后NF-κB核转位情况。5.实时荧光定量RT-PCR及酶联免疫吸附法(ELISA)分析PD98059(ERK抑制剂),SB203580(P38抑制剂),SP600125(JNK抑制剂),PDTC(NF-κB抑制剂)对芍药苷诱导16HBE细胞HBD-2表达及释放的影响。 [结果]1.浓度在100μM以下芍药苷对16HBE细胞无毒。2.芍药苷诱导16HBE细胞HBD-2mRNA表达具有时间和剂量依赖关系。3.芍药苷可显著诱导16HBE细胞ERK,P38MAPK, JNK磷酸化。4.芍药苷可刺激NF-κB核转位。5.PD98059,SB203580,PDTC显著抑制芍药苷诱导的16HBE细胞HBD-2生成及释放,而SP600125的抑制作用不明显。 [结论]芍药苷通过激活16HBE细胞的MAPK通路中的ERK, p38及NF-κB信号途径显著诱导支气管上皮细胞系16HBE表达及释放HBD-2。中药芍药苷通过增加内源性抗感染物质β防御素2表达,增强特异性和非特异性免疫功能,从而发挥抗感染重要作用。
[Abstract]:[objective] to investigate whether paeoniflorin can regulate the expression of HBD-2 and its signal pathway in bronchial epithelial cell line 16HBE. [methods] the activity of 16HBE cells exposed to different concentrations of paeoniflorin was determined by 1.CCK8 assay. Dynamic expression of HBD-2mRNA in 16HBE cells stimulated by Paeoniflorin by Real-time fluorescence quantitative RT-PCR. 3. Western blot (Western Blotting) was used to detect the expression levels of pEK, ERK, P38MAPK, p38MAPK, and JNK in 16HBE cells stimulated by paeoniflorin. 4 cell immunofluorescence assay was used to detect the translocation of NF- 魏 B nucleus in 16HBE cells stimulated by paeoniflorin. The effects of PD98059 (ERK inhibitor), SB203580 (P38 inhibitor) and SP600125 (JNK inhibitor), PDTC (NF- 魏 B inhibitor) on HBD-2 expression and release in 16HBE cells induced by paeoniflorin were analyzed by real-time fluorescence quantitative RT-PCR and enzyme-linked immunosorbent assay (ELISA). [result] 1. Paeoniflorin below 100 渭 M was non-toxic to 16HBE cells. Paeoniflorin induced HBD-2mRNA expression in 16HBE cells in a time-and dose-dependent manner. 3. Paeoniflorin significantly induced phosphorylation of ERK,P38MAPK, JNK in 16HBE cells. Paeoniflorin could stimulate the nuclear translocation of NF- 魏 B. 5.PD98059, SB203580,PDTC significantly inhibited the production and release of HBD-2 in 16HBE cells induced by paeoniflorin, but the inhibitory effect of SP600125 was not obvious. [conclusion] Paeoniflorin can significantly induce the expression of 16HBE and release of HBD-2. from bronchial epithelial cell lines by activating ERK, p38 and NF- 魏 B signaling pathways in the MAPK pathway of 16HBE cells. Paeoniflorin plays an important role in anti-infection by increasing the expression of endogenous anti-infective substance 尾-defensin 2 and enhancing the specific and non-specific immune function.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R56
本文编号：2431708
[Abstract]:[objective] to investigate whether paeoniflorin can regulate the expression of HBD-2 and its signal pathway in bronchial epithelial cell line 16HBE. [methods] the activity of 16HBE cells exposed to different concentrations of paeoniflorin was determined by 1.CCK8 assay. Dynamic expression of HBD-2mRNA in 16HBE cells stimulated by Paeoniflorin by Real-time fluorescence quantitative RT-PCR. 3. Western blot (Western Blotting) was used to detect the expression levels of pEK, ERK, P38MAPK, p38MAPK, and JNK in 16HBE cells stimulated by paeoniflorin. 4 cell immunofluorescence assay was used to detect the translocation of NF- 魏 B nucleus in 16HBE cells stimulated by paeoniflorin. The effects of PD98059 (ERK inhibitor), SB203580 (P38 inhibitor) and SP600125 (JNK inhibitor), PDTC (NF- 魏 B inhibitor) on HBD-2 expression and release in 16HBE cells induced by paeoniflorin were analyzed by real-time fluorescence quantitative RT-PCR and enzyme-linked immunosorbent assay (ELISA). [result] 1. Paeoniflorin below 100 渭 M was non-toxic to 16HBE cells. Paeoniflorin induced HBD-2mRNA expression in 16HBE cells in a time-and dose-dependent manner. 3. Paeoniflorin significantly induced phosphorylation of ERK,P38MAPK, JNK in 16HBE cells. Paeoniflorin could stimulate the nuclear translocation of NF- 魏 B. 5.PD98059, SB203580,PDTC significantly inhibited the production and release of HBD-2 in 16HBE cells induced by paeoniflorin, but the inhibitory effect of SP600125 was not obvious. [conclusion] Paeoniflorin can significantly induce the expression of 16HBE and release of HBD-2. from bronchial epithelial cell lines by activating ERK, p38 and NF- 魏 B signaling pathways in the MAPK pathway of 16HBE cells. Paeoniflorin plays an important role in anti-infection by increasing the expression of endogenous anti-infective substance 尾-defensin 2 and enhancing the specific and non-specific immune function.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R56
【参考文献】
相关期刊论文 前1条
1 王卓;姚婉贞;夏国光;;慢性阻塞性肺部疾病患者血清及痰中β-防御素-2水平及临床意义[J];北京医学;2008年10期
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