摘要
目的:探究氧自由基在纳米细菌(Nanobacteria,NB)导致肾结石损伤过程中的作用。方法:实验分五组:空白对照组、NB组、一水草酸钙(Calcium oxalate monohydrate,COM)组、纳米级羟基磷灰石(Nanograde hydroxuapatite,nHAP)组和四环素(TET+NB)组。将HK-2细胞分别与各组作用物共培养6 h、12 h和24 h后,用透射电子显微镜观察HK-2细胞的形态学变化;超声粉碎细胞后检测Na^+/K^+ATP酶和Ca^2+/Mg^2+ATP酶活性;检测培养液中乳酸脱氢酶(LDH)、过氧化氢(H2O2)和丙二醛(MDA)含量;最后加入COM晶体后用激光共聚焦显微镜观察各组细胞的晶体黏附情况。结果:电镜结果可见,nHAP组和四环素组中HK-2细胞的损伤程度明显低于NB组。nHAP对HK-2细胞酶活性的抑制作用显著低于NB,四环素组细胞在各时间点的酶活性均高于NB组,差异有统计学意义(P<0.05)。各时间点,NB组培养液中H 2O 2和MDA的含量显著高于对照组,差异有统计学意义(P<0.05),nHAP组无明显变化(P>0.05),四环素组显著低于NB组,差异有统计学意义(P<0.05)。随着共培养时间延长,NB组、COM组细胞的晶体黏附量持续增加。当共培养24 h时,nHAP组可观察到少量晶体黏附。各时间点,四环素组中晶体黏附量均显著少于NB组。结论:NB可能通过诱导脂质过氧化反应损伤HK-2细胞,损伤程度和晶体黏附量与NB作用时间成正比,氧自由基物质,可加重过氧化反应,形成恶性循环。四环素可以抑制NB对HK-2细胞的损伤并减少损伤后的晶体滞留。
Objective To explore the role of oxygen free radicals in the process of kidney stone injury caused by nanobacteria.Method The experiment was divided into five groups:blank control group,NB group,calcium oxalate monohydrate(COM)group,nano hydroxyapatite(nHAP)group and tetracycline(TET+NB)group.HK-2 cells were co cultured with each other for 6 h,12 h and 24 h respectively.The morphological changes of HK-2 cells were observed by transmission electron microscope;the activities of Na^+/K^+ ATPase and Ca^2+/Mg^2+ ATPase were detected after ultrasonic comminution;the contents of lactate dehydrogenase(LDH),hydrogen peroxide(H2O2)and malondialdehyde(MDA)in the culture medium were detected;At last,com crystal was added and the crystal adhesion of each group was observed by laser confocal microscope.Results The damage degree of HK-2 cells in nHAP group and tetracycline group was significantly lower than that in NB group.The inhibitive effect of nHAP on HK-2 cells was significantly lower than that of NB,and the enzyme activity of tetracycline group was higher than that of NB group at each time point,the difference was statistically significant(P<0.05).At each time point,the content of H2O2 and MDA in NB group was significantly higher than that in control group(P<0.05),but there was no significant change in nHAP group(P>0.05),while the tetracycline group was significantly lower than that of NB group(P<0.05).With the prolongation of co culture time,the cell adhesion of NB group and COM Group continued to increase.When co cultured for 24 h,a small amount of crystal adhesion was observed in nHAP group.At each time point,the amount of crystal adhesion in the tetracycline group was significantly less than that in the NB group.Conclusion NB may damage HK-2 cells by inducing lipid peroxidation.The degree of injury and the amount of crystal adhesion are directly proportional to the time of NB treatment.Oxygen free radicals can aggravate the peroxidation and form a vicious cycle.Tetracycline can inhibit the damage of NB to HK-2 cells
作者
王敬珅
郝志强
杨恒
汪渊
李永乐
钱彪
郑丽英
冷欣
王艳军
张志强
WANG Jing-shen;HAO Zhiqiang;YANG Heng;无(Shihezi University Medical College,Shihezi 832000,China;Department of Urology,The First Affiliated Hospital of Gannan Medical University,Ganzhou 341000,China;Department of Scientific Research of The First Affiliated Hospital of Gannan Medical College,Ganzhou 341000,China;Department of Urology,The Ninth Division Hospital of Xinjiang Production and Construction Corps,Tacheng 834601,China;Department of Urology,Red Star Hospital,Thirteenth Division,Xinjiang Production and Construction Corps,Hami 839000,China;Department of Urology,The Seventh Division Hospital of Xinjiang Production and Construction Corps,Kuitun,833200,China)
出处
《吉林医学》
CAS
2020年第9期2053-2057,共5页
Jilin Medical Journal
基金
国家自然科学基金[项目编号:81460140]
石河子大学成果转化与技术推广[项目编号:CGZH201810]。
