中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2015年
2期
208-210
,共3页
陈倩%顾健腾%段家翔%袁碧英%鲁开智
陳倩%顧健騰%段傢翔%袁碧英%魯開智
진천%고건등%단가상%원벽영%로개지
再灌注损伤%肾%血清%内皮细胞
再灌註損傷%腎%血清%內皮細胞
재관주손상%신%혈청%내피세포
Reperfusion injury%Kidney%Serum%Endothelial cells
目的 探讨如何建立肾缺血再灌注损伤小鼠血清致肺微血管内皮细胞(PMVECs)损伤模型.方法 培养小鼠PMVECs,测定单层PMVECs标准电阻.待PMVECs生长成单层致密连接状态且达到标准电阻时,采用随机数字表法,将其分为4组(n=3):正常小鼠血清组(NS组)和不同浓度肾缺血再灌注损伤小鼠血清组(IRS5组、IRS10组和IRS20组).换成无血清内皮细胞培养基饥饿处理1h后,NS组上室和下室分别加入含20％正常小鼠血清的内皮细胞培养基0.8和0.2 ml进行培养;IRS5组、IRS10组和IRS20组上室和下室分别加入含5％、10％、20％肾缺血再灌注损伤小鼠血清的内皮细胞培养基0.8和0.2 ml进行培养.4组均在上室中加入100 μg/ml FITC-BSA 100μl作为通透物质.分别于培养3、6、9、12、15、18、21和24 h时,采用荧光白蛋白滤过法检测PMVECs的通透性,以通透系数(Pa)表示.结果 与NS组比较,IRS5组培养12和15 h时Pa升高,IRS10组和IRS20组培养6～24 h时Pa升高(P＜0.05);与IRS5组比较,IRS10组培养21和24 h时Pa升高,IRS20组培养9～24 h时Pa升高(P＜0.05);与IRS10组比较,IRS20组培养15～24 h时Pa升高(P＜0.05).结论 10％和20％肾缺血再灌注损伤小鼠血清均可成功建立PMVECs损伤模型,其中20％血清时损伤更为明显.
目的 探討如何建立腎缺血再灌註損傷小鼠血清緻肺微血管內皮細胞(PMVECs)損傷模型.方法 培養小鼠PMVECs,測定單層PMVECs標準電阻.待PMVECs生長成單層緻密連接狀態且達到標準電阻時,採用隨機數字錶法,將其分為4組(n=3):正常小鼠血清組(NS組)和不同濃度腎缺血再灌註損傷小鼠血清組(IRS5組、IRS10組和IRS20組).換成無血清內皮細胞培養基饑餓處理1h後,NS組上室和下室分彆加入含20％正常小鼠血清的內皮細胞培養基0.8和0.2 ml進行培養;IRS5組、IRS10組和IRS20組上室和下室分彆加入含5％、10％、20％腎缺血再灌註損傷小鼠血清的內皮細胞培養基0.8和0.2 ml進行培養.4組均在上室中加入100 μg/ml FITC-BSA 100μl作為通透物質.分彆于培養3、6、9、12、15、18、21和24 h時,採用熒光白蛋白濾過法檢測PMVECs的通透性,以通透繫數(Pa)錶示.結果 與NS組比較,IRS5組培養12和15 h時Pa升高,IRS10組和IRS20組培養6～24 h時Pa升高(P＜0.05);與IRS5組比較,IRS10組培養21和24 h時Pa升高,IRS20組培養9～24 h時Pa升高(P＜0.05);與IRS10組比較,IRS20組培養15～24 h時Pa升高(P＜0.05).結論 10％和20％腎缺血再灌註損傷小鼠血清均可成功建立PMVECs損傷模型,其中20％血清時損傷更為明顯.
목적 탐토여하건립신결혈재관주손상소서혈청치폐미혈관내피세포(PMVECs)손상모형.방법 배양소서PMVECs,측정단층PMVECs표준전조.대PMVECs생장성단층치밀련접상태차체도표준전조시,채용수궤수자표법,장기분위4조(n=3):정상소서혈청조(NS조)화불동농도신결혈재관주손상소서혈청조(IRS5조、IRS10조화IRS20조).환성무혈청내피세포배양기기아처리1h후,NS조상실화하실분별가입함20％정상소서혈청적내피세포배양기0.8화0.2 ml진행배양;IRS5조、IRS10조화IRS20조상실화하실분별가입함5％、10％、20％신결혈재관주손상소서혈청적내피세포배양기0.8화0.2 ml진행배양.4조균재상실중가입100 μg/ml FITC-BSA 100μl작위통투물질.분별우배양3、6、9、12、15、18、21화24 h시,채용형광백단백려과법검측PMVECs적통투성,이통투계수(Pa)표시.결과 여NS조비교,IRS5조배양12화15 h시Pa승고,IRS10조화IRS20조배양6～24 h시Pa승고(P＜0.05);여IRS5조비교,IRS10조배양21화24 h시Pa승고,IRS20조배양9～24 h시Pa승고(P＜0.05);여IRS10조비교,IRS20조배양15～24 h시Pa승고(P＜0.05).결론 10％화20％신결혈재관주손상소서혈청균가성공건립PMVECs손상모형,기중20％혈청시손상경위명현.
Objective To establish the model of serum-caused damage to pulmonary microvascular endothelial cells (PMVECs) of mice with renal ischemia-reperfusion (I/R) injury.Methods Mice PMVECs were cultured to measure the standard trans-endothelial electrical resistance (TER) in the monolayer of PMVECs.When PMVECs were cultured and arranged in compact monolayer and TER was achieved,they were divided into 4 groups (n =3 each) using a random number table:serum of normal mice group (NS group) and different concentrations (5％,10％ and 20％) of serum of mice with renal I/R injury groups (IRS5 group,IRS10group and IRS20 group).The PMVECs were cultured for 1 h in the serum-free endothelial culture medium.The 0.8 and 0.2 ml culture medium containing 20％ serum of normal mice were then added to the upper and lower chambers,respectively,in group NS.The 0.8 and 0.2 ml culture medium containing 5％,10％ and 20％ serum of mice with renal I/R injury were then added to the upper and lower chambers in IRS5,IRS10 and IRS20 groups,respectively.100 μg/ml FITC-BSA 100 μl was added to the upper chamber in the four groups.At 3,6,9,12,15,18,21 and 24 h of incubation,the PMVEC monolayer permeability (apparent permeability coefficient,Pa) was detected.Results Compared with NS group,the Pa was significantly increased at 12 and 15 h of incubation in IRS5 group,and the Pa was increased at 6-24 h of incubation in IRS10 and IRS20 groups.Compared with IRS5 group,the Pa at 21 and 24 h in IRS10 group and at 9-24 h in IRS20 group were significantly increased.Conclusion Both 10％ and 20％ serum of mice with renal I/R injury can successfully establish the model of damage to PMVECs,and 20％ serum causes a more severe damage.
