摘要
目的观察血管内皮细胞生长因子(VEGF)对内皮细胞血清类黏蛋白(ORM)1表达的影响,探讨转录调控核因子-κB(NF-κB)通路在其中的作用。方法Real-time PCR检测ORM1基因表达。免疫荧光分析P65蛋白核移位。Western blot检测ORM1和P65蛋白表达。结果对照组、低剂量VEGF组、高剂量VEGF组、PDTC组ORM1基因的2^-△△Ct值分别为0.324±0.053、0.252±0.042、0.192±0.034、0.166±0.027,各组间差异有统计学意义(P<0.05)。低剂量VEGF组、高剂量VEGF组、PDTC组细胞核内的P65蛋白表达显著低于对照组。对照组、低剂量VEGF组、高剂量VEGF组、PDTC组细胞核内P65蛋白表达的相对灰度值分别为0.527±0.114、0.316±0.065、0.242±0.038、0.105±0.012,组间比较差异有统计学意义(P<0.05);各组ORM1蛋白表达的相对灰度值分别为0.874±0.142、0.462±0.083、0.305±0.047、0.126±0.014,组间比较差异有统计学意义(P<0.01)。结论VEGF通过抑制NF-κB通路下调ORM1基因、蛋白表达,可能是VEGF致脑水肿的作用机制。
Objective To observe the effect of VEGF on ORM1 expression in endothelial cells via NF-κB pathway,and study the role of NF-κB pathway in it.Methods The expression of ORM1 mRNA was detected by Real-time PCR analysis.The P65 protein nuclear translocation was detected by immunofluorescence.The expression of ORM1 and P65 was determined by Western blot.Results The 2^-△△Ct value of control group,low-dose VEGF group,high-dose VEGF and PDTC group was 0.324±0.053,0.252±0.042,0.192±0.034,and 0.166±0.027,respectively.There were significant differences among the groups(P<0.05).The expression of P65 protein in low-dose VEGF group,high-dose VEGF group and PDTC group was lower than that of control group.The relative gray value of nuclear P65 in control group,low-dose VEGF group,high-dose VEGF group and PDTC group was 0.527±0.114,0.316±0.065,0.242±0.038,and 0.105±0.012,respectively(P<0.05);the relative gray value of ORM1 in the above groups was 0.874±0.142,0.462±0.083,0.305±0.047,and 0.126±0.014,respectively(P<0.01).Conclusion VEGF down-regulates the gene and protein expression of ORM1 by inhibiting NF-κB pathway,which may be the mechanism of brain edema induced by VEGF.
作者
陈琳
周经霞
曾超胜
CHEN Lin;ZHOU Jing-xia;ZENG Chao-sheng(Department of Neurology,Second Affiliated Hospital of Hainan MedicalCollege,Haikou 570311,China)
出处
《实用药物与临床》
CAS
2020年第2期114-117,共4页
Practical Pharmacy and Clinical Remedies
基金
2018年海南省医药卫生科研项目(1801032054A2011)
2018年海南省自然科学基金(818MS149)
