超声联合微泡对内皮细胞一氧化氮合酶磷酸化程度的影响  

Effects of ultrasound combined with microbubble on endothelial nitric oxide synthase phosphorylation

作　　者：吴玉鹏 李岩峰[1] 马逸[1] WU Yu-peng;LI Yan-feng;MA Yi(The People's Hospital of Liaoning Province,Shenyang 110016,Liaoning,China)

机构地区：[1]辽宁省人民医院,辽宁沈阳110016

出　　处：《生物医学工程与临床》2019年第1期14-18,共5页Biomedical Engineering and Clinical Medicine

摘　　要：目的探索超声联合微泡对内皮细胞一氧化氮合酶(e NOS)磷酸化程度的影响。方法将人脐静脉内皮细胞分为3组(对照组、单纯超声组、超声微泡组),超声微泡组给予超声(频率1 MHz,输出功率100 k Pa,占空比20%)联合微泡(500μL)刺激2 min,单纯超声组给予单纯超声(频率1 MHz,输出功率100 kPa,占空比20%)刺激2 min,对照组无任何干预。应用Western blot方法检测磷酸化一氧化氮合酶(S1177)[p-eNOS(S1177)]和总e NOS表达、4’,6-二脒基-2-苯基吲哚(DAPI)染色检测细胞核数目及台盼蓝染色检测3组细胞存活率。结果 3组总e NOS相对表达量分别为1.00±0.09、1.00±0.11、0.99±0.02,差异无统计学意义(P> 0.05)。3组p-eNOS(S1177)相对表达量分别为0.41±0.10、0.49±0.08、0.98±0.05,与对照组和单纯超声组比较,超声微泡组p-eNOS (S1177)表达水平显著升高(P <0.05)。p-eNOS(S1177)与总e NOS的比值亦显著升高(P <0.05)。而3组细胞存活率、细胞核数量及细胞核形态未见明显差异(P> 0.05)。结论超声联合微泡可以促进e NOS磷酸化、p-eNOS(S1177)表达增加,并且不会对内皮细胞的存活产生影响;为临床应用超声联合微泡治疗微血管阻塞提供了理论基础,也为心血管疾病的治疗提供一个新思路。Objective To observe the effects of ultrasound combined microbubble on the expression of endothelial nitric oxide synthase(eNOS)and its phosphorylation at serine 1177[p-eNOS(S1177)]in human umbilical vein endothelial cells(HUVEC).Methods The HUVEC were divided into 3 groups:control group(without any treatment),ultrasound group[ultrasound(frequency1 MHz,output power 100 kPa,duty cycle 20%)simulation for 2 minutes]and ultrasound combined microbubble group[ultrasound(frequency 1 MHz,output power 100 kPa,duty cycle 20%)combined with microbubble(500μL)stimulation for 2 minutes].The expressions of eNOS and p-eNOS(S1177)were detected by Western blotting method.The number of cell nucleus was tested by2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride(DAPI)staining,and cell viability was estimated by trypan blue staining.Results The relative expressions of total eNOS in 3 groups were 1.00±0.09,1.00±0.11,0.99±0.02,respectively,and there was no significant difference(P>0.05).The relative expression levels of p-eNOS(S1177)in 3 groups were 0.41±0.10,0.49±0.08,and 0.98±0.05,respectively.Compared with control group and ultrasound group,the expression level of p-eNOS(S1177)in ultrasound combined microbubble group was significantly increased(P<0.05),the ratios of p-eNOS(S1177)and total eNOS were also significantly increased(P<0.05).There was no significant difference in cell survival rate,nucleus and morphology in 3 groups(P>0.05).Conclusion It is demonstrated that ultrasound combined microbubble could promote endothelial cell eNOS phosphorylation and induce expression of p-eNOS(S1177),which provide theoretic basis for further clinical application of ultrasound combined microbubble in treatment of microvascular occlusion,as well as a novel idea for the treatment of cardiovascular disease.

关 键 词：超声 微泡 内皮细胞 一氧化氮合酶 磷酸化 

分 类 号：R454.3[医药卫生—治疗学]