摘要
目的探究纳米细菌(nanobacteria,NB)损伤人肾小管上皮细胞HK-2并导致晶体滞留的机制以及对该损伤提出防治策略。方法应用肾结石患者尿液培养NB,实验分5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)组和四环素干扰组。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;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组和四环素干扰组中HK-2细胞的损伤程度明显低于NB组。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,四环素干扰组中晶体黏附量均显著少于NB组。结论NB可损伤HK-2细胞,且损伤程度和晶体黏附量与NB作用时间成正比。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 pathological mechanism of nanobacteria(NB)-induced damage to human tubular epithelial HK-2 cells and the subsequent stone formation,so as to put foward prevention&treatment of it.Methods NB was isolated from the urine of patients who were clinically diagnosed as kidney stones.Five groups were established:control group,NB group,calcium oxalate monohydrate(COM)group,nanograde hydroxuapatite(nHAP)group and tetracycline(TET)group.After incubated 6h,12h and 24h,respectively,the cell morphology was observed under transmission election microscope(TEM).The ATPase activity and a series of damage related molecules of cells were detected.Laser scanning confocal microscope(LSCM)was used to observe the crystals adhesion to cells.Results As shown by the observations of TEM,the degree of cell damage of nHAP and TET groups were significantly lower than that of NB group.With the prolongation of co-culture time,the crystal adhesion of cells in NB and COM groups grew continuously.A small account of crystal adhesion was observed in nHAP group after incubated 24h.The number of crystals adhesion in TET group was significantly lower than that in NB group regardless the incubation time.Conclusion NB may damage HK-2 cells by inducing lipid peroxidation.The degree of cell damage and the amount of crystal adhesion were proportional to the incubation time of NB.Tetracycline showed potential to inhibit this process.
作者
王敬珅
王议鹤
郝志强
杨恒
汪渊
李永乐
王勤章
钱彪
WANG Jing-shen;WANG Yi-he;HAO Zhi-qiang(Department of Urology Surgery,the First Affiliated Hospital of Medical College in Shihezi University,Shihezi,Xinjiang 832000,China)
出处
《实用医药杂志》
2020年第1期43-46,50,I0001,共6页
Practical Journal of Medicine & Pharmacy
基金
国家自然科学基金(81460140)
