二氮嗪对新生儿窒息后血清诱导人近曲肾小管上皮细胞损伤的保护作用

Protective Effects of Diazoxide on Injury of Human Renal Tubular Cell Induced by Serum Obtained from Neonates with Asphyxia

目的探讨二氮嗪对窒息后血清诱导人近曲肾小管上皮细胞(HK-2)损伤的保护作用。方法以HK-2为研究对象,以200 mL/L新生儿窒息后血清作为攻击因素处理HK-2细胞。将HK-2细胞分为对照组、窒息组、二氮嗪组。对照组加入100 mL/L胎牛血清培养液;窒息组加入等量200 mL/L新生儿窒息后血清培养液;二氮嗪组将二氮嗪加入等量200 mL/L新生儿窒息后血清培养液,使终浓度为100μmol/L。倒置相差显微镜下观察HK-2细胞的形态学改变并照相,四甲基偶氮唑蓝比色法检测HK-2细胞活力,生物化学法检测细胞培养液LDH漏出率。结果对照组细胞贴壁良好,呈铺路石样,为扁平的多角形,分裂相多,细胞排列紧密,连接成片,数量多。与对照组相比,窒息组细胞受损明显,形态改变,由典型的扁平多角形细胞变为不规则圆形或椭圆形,细胞间空隙加大,连接松散,细胞间可见较多细胞碎片;细胞活力下降,LDH漏出率增加(P<0.05)。与窒息组相比,二氮嗪组细胞损伤状况明显得到改善,细胞生长状况明显好转,形态明显变好;细胞活力增加,LDH漏出率降低(P<0.05),但未恢复至对照组水平。结论二氮嗪具有减轻窒息后血清诱导的HK-2损伤的作用。

Objective To explore the protective effects of diazoxide on injury of human renal tubular cell （ HK - 2 ） induced by serum obtained from neonates with asphyxia. Methods HK - 2 cells was used as the target cell. The attacking concentration of serum obtained from neonates with asphyxia was 200 mL/L. The experiment was designed as 3 groups. HK - 2 cells were divided into control group, asphyxia group,and diazoxide group. Control group ：joined nutrient fluid including 100 mL/L embryo cow blood serum. Asphyxia group ：joined nutrient fluid including the isometric 200 mL/L serum obtained from neonates with asphyxia. Diazoxide group：the diazoxide was joined nutrient including the isometric 200 mL/L serum obtained from neonates with asphyxia fluid. The diazoxide density finally was 100 μmol/L. Then the change of morphology was observed and photographed under inverted microscope ,and the cell viability was measured by methyl thiazolyl tetrazolium method,and the leakage rate oflactate dehydrogenase（LDH） was determined by biochemical methods. Results Under inverted microscopy, HK -2 cells in control group pastes the wall to be good, assumes the paving stone type, into flat polygon, fission many, the cell arrangement was close, connection large expanse,quantity were many. Compared with control group, the HK -2 cell to suffer injury obviously, the shape changed, become the anomalous circular or the ellipse by the model flat polygonal cell, the intercellular space crevice enlarged, the connection was loose, intercellular space obviously many cell fragmented. Living cell quantity reduced obviously, the cell vigor dropped, and the leakage rate of LD14 increased significantly in asphyxia group（ P 〈 0.05 ）. But compared with asphyxia group, the change of morphology of HK -2 was greatly improved, the cell injury decreased obviously, the leakage rate of LDH decreased and viability increased in diazoxide group （ P 〈 0.05 ）, but not to the same as that of control group. Conclusion Diazoxide can relieve HK - 2 injury induced by postasphyxial - serum of neonate.