Lacorte Livia M, Rinaldi Jaqueline C, Justulin Luis A, Delella Flávia K, Moroz Andrei, Felisbino Sérgio L
Univ Estadual Paulista - UNESP, Institute of Biosciences, Department of Morphology, Extracellular Matrix Laboratory, Botucatu, SP, Brazil.
Univ Estadual Paulista - UNESP, School of Pharmaceutical Sciences, Department of Bioprocess and Biotechnology, Cell Culture Laboratory, Araraquara, SP, Brazil.
Biochem Biophys Res Commun. 2015 Feb 20;457(4):538-41. doi: 10.1016/j.bbrc.2015.01.019. Epub 2015 Jan 16.
Matrix metalloproteinases (MMPs) are zinc (Zn(2+)) and calcium (Ca(2+)) dependant endopeptidases, capable of degradation of numerous components of the extracellular matrix. Cadmium (Cd(2+)) is a well known environmental contaminant which could impair the activity of MMPs. In this sense, this study was conducted to evaluate if Cd(2+) intake inhibits these endopeptidases activities at the rat prostate and testicles and if it directly inhibits the activity of MMP2 and MMP9 at gelatinolytic assays when present in the incubation buffer. To investigate this hypothesis, Wistar rats (5 weeks old), were given tap water (untreated, n = 9), or 15 ppm CdCl2 diluted in drinking water, during 10 weeks (n = 9) and 20 weeks (n = 9). The animals were euthanized and their ventral prostate, dorsal prostate, and testicles were removed. These tissue samples were processed for protein extraction and subjected to gelatin zymography evaluation. Additionally, we performed an experiment of gelatin zymography in which 5 μM or 2 mM cadmium chloride (CdCl2) was directly dissolved at the incubation buffer, using the prostatic tissue samples from untreated animals that exhibited the highest MMP2 and MMP9 activities in the previous experiment. We have found that CdCl2 intake in the drinking water led to the inhibition of 35% and 30% of MMP2 and MMP9 (p < 0.05) at the ventral prostate and testis, respectively, in Cd(2+) treated animals when compared to controls. Moreover, the activities of the referred enzymes were 80% and 100% inhibited by 5 μM and 2 mM of CdCl2, respectively, even in the presence of 10 mM of CaCl2 within the incubation buffer solution. These important findings demonstrate that environmental cadmium contamination may deregulate the natural balance in the extracellular matrix turnover, through MMPs downregulation, which could contribute to the toxic effects observed in prostatic and testicular tissue after its exposure.
基质金属蛋白酶(MMPs)是依赖锌(Zn(2+))和钙(Ca(2+))的内肽酶,能够降解细胞外基质的多种成分。镉(Cd(2+))是一种众所周知的环境污染物,可能会损害MMPs的活性。从这个意义上说,本研究旨在评估摄入Cd(2+)是否会抑制大鼠前列腺和睾丸中这些内肽酶的活性,以及在孵育缓冲液中存在时,它是否会在明胶酶解试验中直接抑制MMP2和MMP9的活性。为了研究这一假设,将5周龄的Wistar大鼠分为三组,分别给予自来水(未处理组,n = 9),或在饮用水中稀释的15 ppm CdCl2,持续10周(n = 9)和20周(n = 9)。对动物实施安乐死后,取出其腹侧前列腺、背侧前列腺和睾丸。对这些组织样本进行蛋白质提取,并进行明胶酶谱分析评估。此外,我们进行了一项明胶酶谱实验,将5 μM或2 mM氯化镉(CdCl2)直接溶解在孵育缓冲液中,使用来自未处理动物的前列腺组织样本,这些样本在之前的实验中表现出最高的MMP2和MMP9活性。我们发现,与对照组相比,在Cd(2+)处理的动物中,饮用水中摄入CdCl2分别导致腹侧前列腺和睾丸中MMP2和MMP9的活性抑制35%和30%(p < 0.05)。此外,即使在孵育缓冲溶液中存在10 mM的CaCl2,5 μM和2 mM的CdCl2分别使上述酶的活性抑制80%和100%。这些重要发现表明,环境镉污染可能通过下调MMPs来破坏细胞外基质周转的自然平衡,这可能导致在暴露后前列腺和睾丸组织中观察到的毒性作用。