芍藥苷調(diào)節(jié)人支氣管上皮細胞系16HBE細胞表達HBD-2的分子機制研究
發(fā)布時間：2019-02-28 09:48
【摘要】：[目的]本研究旨在探討芍藥苷能否調(diào)節(jié)支氣管上皮細胞系16HBE表達HBD-2及其相關(guān)的信號通路。 [方法]1.CCK8法測定不同濃度芍藥苷作用下16HBE細胞活性。2.實時熒光定量RT-PCR測試芍藥苷刺激16HBE細胞HBD-2mRNA的動力學表達。3.免疫印跡法(Western Blotting)測定芍藥苷刺激16HBE細胞后p-ERK、ERK、p-P38MAPK、P38MAPK和p-JNK、JNK表達水平。4細胞免疫熒光法檢測芍藥苷刺激16HBE細胞后NF-κB核轉(zhuǎn)位情況。5.實時熒光定量RT-PCR及酶聯(lián)免疫吸附法(ELISA)分析PD98059(ERK抑制劑),SB203580(P38抑制劑),SP600125(JNK抑制劑),PDTC(NF-κB抑制劑)對芍藥苷誘導16HBE細胞HBD-2表達及釋放的影響。 [結(jié)果]1.濃度在100μM以下芍藥苷對16HBE細胞無毒。2.芍藥苷誘導16HBE細胞HBD-2mRNA表達具有時間和劑量依賴關(guān)系。3.芍藥苷可顯著誘導16HBE細胞ERK,P38MAPK, JNK磷酸化。4.芍藥苷可刺激NF-κB核轉(zhuǎn)位。5.PD98059,SB203580,PDTC顯著抑制芍藥苷誘導的16HBE細胞HBD-2生成及釋放,而SP600125的抑制作用不明顯。 [結(jié)論]芍藥苷通過激活16HBE細胞的MAPK通路中的ERK, p38及NF-κB信號途徑顯著誘導支氣管上皮細胞系16HBE表達及釋放HBD-2。中藥芍藥苷通過增加內(nèi)源性抗感染物質(zhì)β防御素2表達,增強特異性和非特異性免疫功能,從而發(fā)揮抗感染重要作用。
[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.
【學位授予單位】：南京醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R56
本文編號：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.
【學位授予單位】：南京醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R56
【參考文獻】
相關(guān)期刊論文 前1條
1 王卓;姚婉貞;夏國光;;慢性阻塞性肺部疾病患者血清及痰中β-防御素-2水平及臨床意義[J];北京醫(yī)學;2008年10期
，本文編號：2431708
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