丹参素干预体外低温保存大鼠肝脏血红素氧合酶1的表达

Danshensu interferes in heme oxygenase-1 expression during in vitro liver cold preservation

背景:研究发现丹参能够降低诱导型一氧化氮合酶mRNA的表达,减少一氧化氮的产生,抑制肿瘤坏死因子、白细胞介素等炎性因子的分泌,具有抗氧化作用。丹参素作为丹参的重要单体成分,是否具有相同的作用?目的:验证丹参素干预大鼠肝脏血红素氧合酶1的表达,以及对体外肝脏低温保存肝脏的保护。方法:建立大鼠肝脏低温灌注保存模型。分为3组,对照组术前腹腔注射生理盐水,术中用乳酸林格液灌注;实验组术前腹腔注射生理盐水,术中用丹参素+乳酸林格液灌注;抑制剂组:术前腹腔注射锌原朴啉,术中用丹参素+乳酸林格液灌注。保存0,1,3,6h,分别RT-PCR检测血红素氧合酶1mRNA及蛋白表达的情况,检测细胞线粒体钙离子含量及钙离子ATP酶活性,电镜观察各组肝细胞、线粒体形态改变,光镜观察肝细胞、肝小叶形态改变。结果与结论:实验组肝血红素氧合酶1mRNA及蛋白的表达水平明显比其他两组高(P<0.05)。实验组的肝细胞线粒体钙离子含量明显较其他两组低,Ca2+-ATP酶活性较其他两组高。结果提示,丹参素灌注保存液可以诱导大鼠肝脏中血红素氧合酶1的过表达,延长肝脏低温保存时间。

BACKGROUND：Previous studies have demonstrated that salvia miltiorrhiza can decrease inducible nitric oxide synthase（iNOS） mRNA expression,reduce nitrogen monoxidum production,inhabit tumor necrosis factor and interleukin secretion,as well as antioxidation.As a monomer component of salvia miltiorrhiza,whether Dasensu has the similar effects remain poorly understood.OBJECTIVE：To verify the intervention of Danshensu on heme oxygenase-1（HO-1） expression protection on liver cold preservation in vitro.METHODS：Rats were prepared for models of liver hypothermic perfusion preservation and divided into 3 groups.Rats in the control group were intraperitoneal injected physiological saline preoperatively and infused by Ringer lactate solution;those in the experimental group were received intraperitoneal injection of physiological saline preoperatively and infused by Danshensu plus infused by Ringer lactate solution in the surgery;those in the inhibitor group were intraperitoneal injected original zinc porphyrin preoperatively and infused by Danshensu plus infused by Ringer lactate solution in the surgery.HO-1mRNA and protein expression was determined by RT-PCR;Mitochondrial calcium content and calcium ions ATP activity were detected,and the hepatocyte and mitochondrial morphological changes were observed under a electron microscopy,and the morphological changes of liver cells and lobular were observed under a light microscope.RESULTS AND CONCLUSION：Hepatic HO-1 mRNA and protein levels of the experimental group were significantly higher than that of the other groups（P 0.05）.Compared with the other groups,the liver mitochondrial calcium content was notably lower in the experimental group,but the Ca2＋-ATP activity was greater.The results reveled that,Danshensu perfusion fluid can induce over-expression of HO-1 and prolong the liver cold preservation time.