天香丹对冠脉微循环障碍大鼠Nrf2/ARE信号通路表达的影响

Effect of Tianxiangdan on Nrf2/ARE Signaling Pathway Expression in Rats with Coronary Microcirculation Dysfunction

目的探究天香丹对冠脉微循环障碍大鼠核转录因子NF-E2相关因子2/抗氧化反应元件(Nrf2/ARE)信号通路相关因子mRNA及蛋白表达的影响。方法36只SPF级SD大鼠随机分为空白对照组、模型组、尼可地尔组、天香丹组,每组9只。除空白对照组外,其余大鼠开胸后左心室内注射月桂酸钠建立冠脉微循环障碍大鼠模型。各组大鼠分别给予天香丹[0.9408(大鼠体重)^2/3 g/d]、尼可地尔[0.7840(大鼠体重)^2/3mg/d]、生理盐水灌胃28天,每只大鼠给药体积均为2 mL/d。HE染色观察大鼠心肌形态学变化,RT-PCR及Western Blot法观察各组大鼠心肌中Nrf2/ARE信号通路核转录因子NF-E2相关因子2(Nrf2)、Kelch样环氧氯丙烷相关蛋白-1(Keap1)、醌氧化还原酶1(NQO1)、氧化氢酶(CAT)、血红素加氧酶-1(HO-1)mRNA及蛋白的表达变化。结果HE染色显示天香丹可减轻冠脉微循环障碍大鼠微血栓程度。与空白对照组比较,模型组大鼠Nrf2、NQO1、CAT mRNA表达降低(P<0.05),Keap1 mRNA及蛋白表达均降低(P<0.05);与模型组比较,尼可地尔组Nrf2 mRNA及蛋白表达量明显升高(P<0.05),CAT mR-NA表达升高(P<0.05),HO-1 mRNA表达降低(P<0.05);天香丹组Nrf2、NQO1、CAT、HO-1 mRNA及蛋白表达升高(P<0.05),Keap1 mRNA表达升高(P<0.05);与尼可地尔组比较,天香丹组Nrf2、Keap1、NQO1、CAT、HO-1 mRNA表达升高(P<0.05),CAT蛋白表达升高(P<0.05)。结论天香丹改善冠脉微循环障碍机制可能与调控Nrf2/ARE信号通路有关。

Objective To research the effect of Tianxiangdan(TXD)on coronary microcirculation dysfunction rat nuclear factor erythroid-2-related factor 2/antioxidant responsive element(Nrf2/ARE)signal pathway related factor mRNA and protein expression.Methods Totally 36 SPF SD rats were randomly divided into blank control group,model group,Nicorandil group,and TXD group,9 rats in each group.Except the rats in the blank control group,the rest rats were injected with sodium laurate into their left ventricle after thoracotomy,thus establishing a rat model of coronary microcirculation dysfunction.The rats in each group were administered intragastrically with TXD[0.9408(kg)^2/3 g/d],Nicorandil[0.7840(kg)^2/3 mg/d],and normal saline for 28 days.The volume of administration was 2 mL/d for per rat.HE staining method was used to observe the morphological changes of rat myocardium.The Nrf2/ARE signaling pathway nuclear factor NF-E2 related factor 2(nuclear factor erythroid-2-related factor 2,Nrf2),Kelch-like ECH-associated protein-1(Keap1),NAD(P)H:quinone oxidoreductase 1(NQO1),catalase(CAT),heme oxygenase-1(HO-1)mRNA and protein expression were detected by RT-PCR and Western Blot respectively.Results According to HE staining method,TXD could reduce the degree of micro thrombosis in coronary microcircu lation dysfunction rats.Compared with the blank control group,the expression of Nrf2,NQO1,CAT mRNA in the model group reduced(P<0.05),and the expression of Keap1 mRNA and protein decreased(P<0.05).Com-pared with the model group,the expression of Nrf2 mRNA and protein in the Nicorandil group was significantly increased(P<0.05).The expression of CAT mRNA increased(P<0.05),and the expression of HO-1 mRNA was decreased(P<0.05).In TXD group,Nrf2,NQO1,CAT,HO-1 mRNA and protein expression increased(P<0.05),the expression of Keap1 mRNA increased(P<0.05).Compared with the Nicorandil group,the expression of Nrf2,Keap1,NQO1,CAT,HO-1 mRNA in TXD group increased(P<0.05),and the expression of CAT protein increased(P<0.05).Conclusion TXD's mechanism for improving coronary microcirculation dysfunction may be as-sociated with the regulation of the Nrf2/ARE signal pathway.