摘要
目的研究银杏叶提取物(EGb)对新生儿窒息后血清攻击人近曲肾小管上皮细胞(HK-2)损伤模型的影响。方法以人近曲肾小管上皮细胞(HK-2)为研究对象,200ml/L窒息血清作为攻击因素。实验共分为两阶段,均设对照组、模型组和EGb干预组(EGb预处理24h)。第一阶段EGb干预组设立5个质量浓度亚组。观察三组细胞形态学,乳酸脱氢酶(LDH)漏出率和细胞存活(MTT法)变化。第二阶段以第一阶段选出的EGb最适质量浓度预处理细胞,观察三组NF-κB p65蛋白(免疫组化)和抑制亚基I-κBα(western blot)的变化。结果与对照组相比,模型组细胞形态发生改变,LDH漏出率增加,细胞活力降低(P〈0.05);与模型组比较,EGb预处理组细胞形态明显得到改善,LDH漏出率降低,细胞活力增加(P〈0.05),在5—50mg/L保护效应逐渐增加,50mg/L时保护效果最强,而〉50mg/L时保护效应反而下降。与对照组相比,模型组细胞NF-κB p65蛋白表达明显增加,I-κBα量减少(P〈0.05);与模型组比较,EGb干预组细胞NF-κB p65蛋白表达明显减少,I-κBα量增加(P〈0.05)。结论EGb具有减轻窒息血清所致HK-2细胞损伤的作用,其作用可能与抑制窒息血清所致NF—κB激活有关。
Objective To investigate the effects of Ginkgo biloba extract(EGb) on injury of renal tubular cells (HK- 2) induced by postasphyxial-serum of neonate. Methods Human renal proximal tubular eell line HK-2 cells were taken as subjects ,the serum of neonates 1 day after asphyxia, which concentration were 200 ml/L (volume fraction) ,were applied as attacking factor. The experiment was derided into two stages, each stage include control group, model group and EGb group ( pretreatment with EGb for 24 h). In the first stage, the EGb group had fivo subgroups. The injury of morphology were observed under inverted microscope. The cell viability was measured by MTT methods. The leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods. In the second stage, the expression of NF-κB p65 and I-κBα was detected by immunohistochemistry and Western blot respectively. Results In the first stage, compared with control group, cell morphous changed, leakage rate of LDH increased and cell survival rate decreased in model group, the variance was of significance (P 〈 0. 05 ) ; Compared with model group, however, cell morphous improved dramatically, leakage rate of LDH decreased and cell survival rate increased in EGb group, the variance was of significance (P 〈 0.05). The protection effect increased gradually when concentration of EGb was in the range of 5 - 50 rag/L, the protection effect was the most powerful at 50 mg/L, while it decreased when the concentartion was over 50 mg/L. In the second stage,the expression of NF-κB increased and the amout of I-κBα decreased in model group ,when compared with the control group, the variance was of significance ( P 〈 0.05). The expression of NF-κB decreased and the amout of I-κBα increased in EGb group ,when compared with the model group, the variance was of significance ( P 〈 0.05). Conclusion EGb could provide the protective effects on the injury of renal tubular ceils induced by postasphyxial-serum of neonate.
出处
《山东医药》
CAS
北大核心
2009年第49期9-12,共4页
Shandong Medical Journal
基金
四川省杰出青年学科带头人培养基金(04ZQ026-033)
四川省中医管理局重点科研基金(200239)
关键词
银杏叶提取物
新生儿
窒息
血清
肾损伤
Ginkgo biloba extract
newborn
asphyxia
serum
renal injury