CAJ | 학술논문

目的探讨早产儿氧暴露后,患儿外周血单个核细胞(PBMCs)内烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶亚单位p47phox调节细胞内活性氧(ROS)升高的机制.方法胎龄＜32周需吸氧早产儿根据吸入氧体积分数(FiO2)不同分为3组:FiO2＜30％为低氧组、FiO2在30％～ 40％为中氧组、FiO2＞ 40％为高氧组.同期未吸氧的＜32周早产儿为对照组.氧疗48 h后,各组经桡动脉采血3 mL,分离PBMCs及血清,应用激光共聚焦显微镜检测PBMCs内ROS生成量,硫代巴比妥酸比色法检测血清丙二醛(MDA)水平,免疫荧光检测p47phox在细胞内定位及p47phox的活化率.结果早产儿氧暴露后,随着FiO2的升高,ROS与MDA逐渐升高,p47phox由胞质向胞膜移位的细胞数增多,p47phox的移位率也在增加;与对照组相比,余3组ROS明显升高,差异有统计学意义(q =4.48、6.5、16.22,P均＜0.05);MDA水平显著增加,差异有统计学意义(q=5.08、8.22、12.76,P均＜0.05);p47phox的活化率比较差异也具有显著性差异(x2=134.008,P＜0.05);与中氧组相比,高氧组ROS和MDA显著升高,差异有统计学意义(q=15.03、4.53,P均＜0.05);p47phox的活化率明显增加,差异有统计学意义(x2=19.26,P＜0.05).结论早产儿氧暴露后,p47phox可能通过向胞膜移位来调节PBMCs内NADPH氧化酶源性活性氧升高.
목적 탐토조산인양폭로후,환인외주혈단개핵세포(PBMCs)내연선알선표령이핵감산린산(NADPH)양화매아단위p47phox조절세포내활성양(ROS)승고적궤제.방법 태령＜32주수흡양조산인근거흡입양체적분수(FiO2)불동분위3조:FiO2＜30％위저양조、FiO2재30％～ 40％위중양조、FiO2＞ 40％위고양조.동기미흡양적＜32주조산인위대조조.양료48 h후,각조경뇨동맥채혈3 mL,분리PBMCs급혈청,응용격광공취초현미경검측PBMCs내ROS생성량,류대파비타산비색법검측혈청병이철(MDA)수평,면역형광검측p47phox재세포내정위급p47phox적활화솔.결과 조산인양폭로후,수착FiO2적승고,ROS여MDA축점승고,p47phox유포질향포막이위적세포수증다,p47phox적이위솔야재증가;여대조조상비,여3조ROS명현승고,차이유통계학의의(q =4.48、6.5、16.22,P균＜0.05);MDA수평현저증가,차이유통계학의의(q=5.08、8.22、12.76,P균＜0.05);p47phox적활화솔비교차이야구유현저성차이(x2=134.008,P＜0.05);여중양조상비,고양조ROS화MDA현저승고,차이유통계학의의(q=15.03、4.53,P균＜0.05);p47phox적활화솔명현증가,차이유통계학의의(x2=19.26,P＜0.05).결론 조산인양폭로후,p47phox가능통과향포막이위래조절PBMCs내NADPH양화매원성활성양승고.
Objective To explore the mechanism for the increase in reactive oxygen species regulated by p47phox of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunit in peripheral blood mononuclear cells (PBMCs) after oxygen therapy in premature infants.Methods According to different volume fractions of oxygen,premature infants less than 32 weeks were divided into 3 groups:fractional concentration of inspired oxygen (FiO2) ＜ 30％ was low concentration oxygen group,FiO2 between 30％ and 40％ as middle concentration oxygen group,and FiO2 ＞ 40％ as high concentration oxygen group.Premature infants less than 32 weeks without oxygen was control group.After 48 h,3 mL blood was collected via radial artery from each group,PBMCs and serum were separated.Then intracellular reactive oxygen species (ROS) by confocal laser scanning microscopy,malondialdehyde (MDA) within serum by thiobarbituric acid colorimetric,and the location and activation rate of p47phox through immunofluorescence.Results After premature infants were exposed to oxygen,as the oxygen volume fraction was increasing,ROS and MDA gradually rised.More PBMCs with p47phox translocated to membrane,then the translocation rate of p47phox also increased.Compared with the control group,ROS were significantly higher(q =4.48,6.5,16.22,all P ＜ 0.05) among the other 3 groups ; MDA significantly increased as well(q =5.08,8.22,12.76,all P ＜ 0.05) ; the activation rate of p47phox also had significant differences (x2 =134.008,P ＜ 0.05);compared with the middle concentration oxygen group,the high concentration oxygen group had higher ROS and MDA(q =15.03,4.53,all P ＜ 0.05) ; the activation rate of p47phox increased significantly(x2 =19.26,P ＜ 0.05).Conclusions After oxygen exposure,p47phox translocated to membrane may regulate the NADPH oxidase-derived ROS increase in extremely premature infants.