Renal effects of metallothionein induction by zinc in vitro and in vivo.
Renal effects of metallothionein induction by zinc in vitro and in vivo.
复制标题
DOI：
10.1186/s12882-017-0503-z
复制
发表时间：
2017-03-16
期刊：
影响因子：
2.3
通讯作者：
Kimmel M
中科院分区：
文献类型：
--
作者：
Schanz M;Schaaf L;Dippon J;Biegger D;Fritz P;Alscher MD;Kimmel M
金属硫蛋白（MTT）是一种内源性抗氧化剂，可由锌（Zn）和缺血诱导。在肾脏中，MTT表达增加在糖尿病和缺氧中发挥假定的保护作用。我们的目的是进一步研究MTT在体外和体内锌和缺氧的影响下的行为。MTT的表达测定在体外培养的近端肾小管细胞（LCC-PK 1）的免疫组织化学和实时荧光PCR孵育后，在缺氧和非缺氧条件下与浓度增加的锌。此外，在54名患者中进行了体内研究，以研究通过Zn的MTT诱导。这是一项使用安慰剂、锌和N-乙酰半胱氨酸预防造影剂诱导肾病的前瞻性、随机、双盲试验的子研究。在p.o.之前和之后2天获得血液样品。用或不用Zn（60 mg）处理。采用ELISA法检测细胞MTT值，评价锌对细胞增殖的影响。对于体内分析，我们相应地考虑了MTT与基线MTT的比值（MTT 1/MTT 0）和eGFR的比值（eGFR 1/eGFR 0）。 体外定量免疫组织化学分析（IHC）和实时PCR显示，随着锌水平的增加，（5、10和15 μg/ml）导致MTTs进行性增加：IHC的中值（IQR）表达也从0.10（0.09-0.12）、0.15（0.12-0.18）、0.25（0.25-0.27）、0.59（0.48-0.70）（p < 0.0001）逐渐增加。PCR表达中位数（IQR）：0.59（0.51-1.72）、1.62（1.38-4.70）、3.58（3.06-10.42）和10.81（9.24-31.47）（p < 0.0001）。与此相反，缺氧并没有改变MTT-水平在体外（p > 0.05）。 在体内没有显着差异（p = 0.96）发生后，锌给药2天的MTT水平相比，没有锌摄入。然而，在锌给药的情况下，MTT（MTT 1/MTT 0）和eGFR（eGFR 1/eGFR 0）之间存在显著相关性（rho = −0.49; 95%-CI：−0.78至−0.03; p = 0.04）。我们发现，锌诱导MTTs在体外，而缺氧没有显着的影响。与此相反，没有显着增加MTTs后，在体内施用锌检测。然而，在Zn给药的情况下，体内MTT和eGFR之间存在显著的负相关性，这可能表明MTTs在肾功能降低的情况下具有保护作用，这可能受到Zn的影响。ClinicalTrials.gov标识符：NCT 00399256。2006年11月13日进行了追溯登记。
Metallothionein (MTT) is an endogenous antioxidant that can be induced by both zinc (Zn) and ischemia. In kidneys, increased MTT expression exerts a putative protective role in diabetes and hypoxia. Our goal was to further investigate the behavior of MTT under the influence of Zn and hypoxia in vitro and in vivo. MTT expression was measured in vitro in cell cultures of proximal tubular cells (LCC-PK1) by immune-histochemistry and real-time PCR after incubation with increasing concentrations of Zn under hypoxic and non-hypoxic conditions. In addition, in vivo studies were carried out in 54 patients to study MTT induction through Zn. This is a sub-study of a prospective, randomized, double-blind trial on prevention of contrast-media-induced nephropathy using Placebo, Zn and N-Acetylcysteine. Blood samples were obtained before and after 2 days p.o. treatment with or without Zn (60 mg). ELISA-based MTT level measurements were done to evaluate the effects of Zn administration. For in vivo analysis, we considered the ratio of MTT to baseline MTT (MTT1/MTT0) and the ratio of eGFR (eGFR1/eGFR0), correspondingly. In vitro quantitative immuno-histochemical analysis (IHC) and real-time PCR showed that at increasing levels of Zn (5, 10, and 15 μg/ml) led to a progressive increase of MTTs: Median (IQR) expression of IHC also increased progressively from 0.10 (0.09–0.12), 0.15 (0.12–0.18), 0.25 (0.25–0.27), 0.59 (0.48–0.70) (p < 0.0001). Median (IQR) expression of PCR: 0.59 (0.51–1.72), 1.62 (1.38–4.70), 3.58 (3.06–10.42) and 10.81 (9.24–31.47) (p < 0.0001). In contrast, hypoxia did not change MTT-levels in vitro (p > 0.05). In vivo no significant differences (p = 0.96) occurred in MTT-levels after 2 days of Zn administration compared with no Zn intake. Nevertheless, there was a significant correlation between MTT (MTT1/MTT0) and eGFR (eGFR1/eGFR0) in case of Zn administration (rho = −0.49; 95%-CI: −0.78 to −0.03; p = 0.04). We found that Zn did induce MTTs in vitro, whereas hypoxia had no significant impact. In contrast, no significant increase of MTTs was detected after in vivo administration of Zn. However, there was a significant negative correlation between MTT and eGFR in vivo in case of Zn administration, this could indicate a protective role of MTTs in a setting of reduced kidney function, which is possibly influenced by Zn. ClinicalTrials.gov Identifier: NCT00399256. Retrospectively registered 11/13/2006.