| 左右,赵庆锁,罗史科,等.血管紧张素Ⅱ调控基质金属蛋白酶 9的表达在蛛网膜下腔出血发病中的机制研究[J].安徽医药,2020,24(3):473-477. |
| 血管紧张素Ⅱ调控基质金属蛋白酶 9的表达在蛛网膜下腔出血发病中的机制研究 |
| Mechanism of AngⅡ regulating MMP?9 expression in pathogenesis of SAH |
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| DOI：10.3969/j.issn.1009?6469.2020.03.013 |
| 中文关键词: 蛛网膜下腔出血 /病因学 基因表达调控，酶学 血管紧张素 Ⅱ 基质金属蛋白酶 9 p38丝裂原活化蛋白激酶类 酶联免疫吸附测定 印迹法，蛋白质 |
| 英文关键词: Subarachnoid hemorrhage/etiology Gene expression regulation,enzymologic Angiotensin Ⅱ Matrix metallopro? teinase?9 p38 Mitogen?activated protein kinases Enzyme?linked immunosorbent assay Blotting,western |
| 基金项目:深圳市盐田区科技计划项目(20170315) |
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| 中文摘要: |
| 目的 探讨血管紧张素 Ⅱ(AngⅡ)调控基质金属蛋白酶 9(MMP?9)的表达在蛛网膜下腔出血( SAH)发病中的机制。方法人脑微血管内皮细胞( HBMEC)株分为 AngⅡ组、 AngⅡ+SB203580组和对照组。其中 AngⅡ组以 100 μg/L浓度 AngⅡ处理 90 min、2h、4h、6h、12 h后取细胞上清液进行相关指标检测； AngⅡ+SB203580组以 5 μΜ的 SB203580处理 20 min后以 100 μg/L浓度 AngⅡ处理 90 min、2h、4h、6h、12 h后取细胞上清液进行相关指标检测；对照组加入 RPMI?1640培养液培养 90 min、2h、4h、6h、12 h后取细胞上清液进行相关指标检测。以酶联免疫吸附法( ELISA)检测各组细胞上清液 MMP?9水平，以实时荧光定量 PCR和蛋白质印迹法检测各组细胞 p38丝裂原活化蛋白激酶( p38 MAPK)的 mRNA和蛋白表达水平。结果并与对照组比较， AngⅡ组细胞上清液 MMP?9［处理 12 h:(4.21±1.36)μg/L比( 5.81±1.72)μg/L，P＜0.01］水平升高， AngⅡ+ SB203580组细胞上清液 MMP?9［处理 12 h:(4.21±1.36)μg/L比( 3.64±1.45)μg/L，P＜0.01］水平降低( P＜0.05)。与 AngⅡ组比较， AngⅡ+SB203580组细胞上清液 MMP?9水平降低( P＜0.001)。与对照组比较， AngⅡ组细胞 p38 MAPK的 mRNA［处理 12 h:(0.422±0.057)比( 0.538±0.071)，P＜0.05)］和蛋白表达水平［( 0.452±0.105)比( 0.635±0.133)P＜0.001］升高， AngⅡ+ SB203580组细胞 p38 MAPK的 mRNA［处理 12 h:(0.422±0.057)比( 0.375±0.066)，P＜0.05］和蛋白表达，水平［(0.452±0.105)比(0.276±0.081)P＜0.001］降低( P＜0.05)。与 AngⅡ组比较， AngⅡ+SB203580组细胞 p38 MAPK的 mRNA和蛋白表达水平降低( P＜0.001)。结论，AngⅡ可能通过激活 p38 MAPK信号通路上调 MMP?9表达从而促进 SAH发生发展。 |
| 英文摘要: |
| Objective To investigate the mechanism of angiotensin Ⅱ(AngⅡ)regulating the expression of matrix metalloprotein? ase?9(MMP?9)in the pathogenesis of subarachnoid hemorrhage(SAH).Methods Human brain microvascular endothelial cell(HBMEC)lines were divided into the AngⅡ group，the AngⅡ +SB203580 group and the control group.The AngⅡ group was treat? ed with 100 μg/L AngⅡ for 90 minutes，2 hours，4 hours，6 hours，12 hours，then the cells supernatant was taken for the relevantindicators detection.The AngⅡ +SB203580 group was treated with 5 μΜ SB203580 for 20 minutes and then treated with 100 μg/LAngⅡ for 90 minutes，2 hours，4 hours，6 hours，12 hours，then the cells supernatant was taken for the relevant indicators detection.The control group was added with RPMI?1640 medium for 90 min，2h，4h，6 h and 12 h，and then cell supernatant was taken fordetection of related indexes.The levels of MMP?9 in cell supernatant were detected by ELISA method，and the expression of p38 mi? togen activated protein kinase(p38 MAPK)messenger ibonucleic acid(mRNA)and protein were detected by real?time quantita? tive polymerase chain reaction and Western Blot.Results Compared with the control group，the level of MMP?9［intervention 12 h:(4.21±1.36)μg/L vs.(5.81±1.72)μg/L，P＜0.001］in the supernatant of AngⅡ group increased，and MMP?9［intervention 12 h:(4.21±1.36)μg/L vs.(3.64±1.45)μg/L，P＜0.01］in the supernatant of AngⅡ +SB203580 group decreased(P＜0.001).Compared with the AngⅡgroup，the level of MMP?9 in supernatant of AngⅡ+SB203580 group decreased(P＜0.05).Compared with the con? trol group，the mRNA［intervention 12 h:(0.422±0.057)vs.(0.538±0.071)，P＜0.05］and protein expression［( 0.452±0.105)vs.(0.635±0.133)，P＜0.001］of p38 MAPK increased in AngⅡ group cells，and the mRNA［intervention 12 h:(0.422±0.057)vs.(0.375±0.066)，P＜0.05］and protein expression［( 0.452±0.105)vs.(0.276±0.081)，P＜0.001］of p38 MAPK decreased in Ang Ⅱ +SB203580 group cells(P＜0.05).Compared with the AngⅡ group，the mRNA and protein expression of p38 MAPK in AngⅡ +SB203580 group decreased(P＜0.001).Conclusion AngⅡ may promote the development of SAH by activating the p38 MAPKsignaling pathway and upregulating the MMP?9 expression. |
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