Muniz Jaqueline J, Leite Letícia N, Lacchini Riccardo, Tanus-Santos José E, Tirapelli Carlos R
a Escola de Enfermagem de Ribeirão Preto, DEPCH, Universidade de São Paulo (USP), Ribeirão Preto, SP, Brazil.
b Departamento de Farmacologia, Faculdade de Medicina de Ribeirão Preto, USP, Ribeirão Preto, SP, Brazil.
Can J Physiol Pharmacol. 2018 Mar;96(3):266-274. doi: 10.1139/cjpp-2017-0082. Epub 2017 Aug 18.
We evaluated the effects of ethanol consumption on the mitogen-activated protein kinases (MAPK) and metalloproteinases (MMP) pathways in the rat cavernosal smooth muscle (CSM). Male Wistar rats were treated with ethanol (20% v/v) for 6 weeks. Quantitative real-time polymerase chain reaction experiments showed that ethanol consumption did not alter mRNA levels of p38MAPK, SAPK/JNK, ERK1/2, MMP-2, or MMP-9 in the rat CSM. Western immunoblotting experiments revealed decreased protein expression of p38MAPK and phosphorylation of SAPK/JNK in the CSM from ethanol-treated rats. Additionally, ethanol consumption decreased the expression of MMP-2. Functional assays showed that SP600125, an inhibitor of SAPK/JNK, prevented the increase in endothelin (ET)-1-induced contraction in the CSM from ethanol-treated rats. Treatment with ethanol decreased MMP-2 activity, but did not change net MMP activity in the rat CSM. Ethanol consumption increased the circulating levels of MMP-2, MMP-9, and TIMP-2 as well as the MMP-9/TIMP-1 ratio. The major finding of our study is that ethanol consumption down-regulates both MAPK and MMP pathways in the rat CSM, whereas it increases the circulating levels of MMP-9. Additionally, we found that SAPK/JNK plays a role in ethanol-induced increase on ET-1 contraction in the isolated rat CSM.
我们评估了乙醇摄入对大鼠海绵体平滑肌（CSM）中丝裂原活化蛋白激酶（MAPK）和金属蛋白酶（MMP）信号通路的影响。雄性Wistar大鼠用乙醇（20% v/v）处理6周。定量实时聚合酶链反应实验表明，乙醇摄入并未改变大鼠CSM中p38MAPK、应激激活蛋白激酶/应激活化蛋白激酶（SAPK/JNK）、细胞外信号调节激酶1/2（ERK1/2）、基质金属蛋白酶-2（MMP-2）或基质金属蛋白酶-9（MMP-9）的mRNA水平。蛋白质免疫印迹实验显示，乙醇处理大鼠的CSM中p38MAPK的蛋白表达降低，SAPK/JNK的磷酸化水平降低。此外，乙醇摄入降低了MMP-2的表达。功能分析表明，SAPK/JNK抑制剂SP600125可阻止乙醇处理大鼠CSM中内皮素（ET）-1诱导的收缩增加。乙醇处理降低了大鼠CSM中MMP-2的活性，但未改变净MMP活性。乙醇摄入增加了循环中MMP-2、MMP-9和金属蛋白酶组织抑制因子-2（TIMP-2）的水平以及MMP-9/TIMP-1的比值。我们研究的主要发现是，乙醇摄入下调了大鼠CSM中的MAPK和MMP信号通路，而增加了循环中MMP-9的水平。此外，我们发现SAPK/JNK在乙醇诱导的离体大鼠CSM中ET-1收缩增加中起作用。
