白藜芦醇对奥沙利铂所致大鼠背根神经节TRP通道影响的作用及机制研究

Effects of Resveratrol on Transient Receptor Potential Channels in Dorsal Root Ganglion of Rats Induced by Oxaliplatin and Its Mechanism

目的探讨白藜芦醇对奥沙利铂所致大鼠背根神经节瞬时受体电位(Transient receptor potential,TRP)通道蛋白TRPV1和TRPM8的影响及其可能的机制。方法将雄性SD大鼠随机分为正常对照组,模型对照组,白藜芦醇低、中、高剂量组。除正常对照组外,其他各组均腹腔注射奥沙利铂4 mg·kg^-1,白藜芦醇各剂量组同时分别灌胃不同剂量(50、100、200 mg·kg^-1)的白藜芦醇。于规定时间处死大鼠,以qPCR和Western Blot法检测L4-5背根神经节TRPV1和TRPM8的表达水平。以Western Blot法检测蛋白激酶A(PKA)和蛋白激酶Cε(PKCε)的磷酸化水平以及蛋白酶激活受体2(PAR2)的表达水平。结果与正常对照组比较,模型对照组大鼠L4-5背根神经节TRPV1和TRPM8 mRNA及蛋白表达水平均明显升高(P<0.05,P<0.01),p-PKA、p-PKCε以及PAR2蛋白表达水平亦明显升高(P<0.05,P<0.01);而白藜芦醇不仅可明显降低L4-5背根神经节TRPV1和TRPM8 m RNA及蛋白的表达水平(P<0.05,P<0.01),同时还可明显抑制p-PKA、p-PKCε以及PAR2的蛋白表达水平(P<0.05)。结论白藜芦醇改善奥沙利铂所致大鼠周围神经毒性的作用机制可能与降低TRPV1、TRPM8表达,抑制PAR2-PKA和PAR2-PKCε的活化有关。

Objective To explore the effects and possible mechanism of resveratrol on transient receptor potential(TRP)proteins TRPV1 and TRPM8 in dorsal root ganglion in rats induced by oxaliplatin. Methods Male SD rats were randomly divided into control group,model group,the low-,medium-and high-dose of resveratrol groups(50,100 and 200 mg·kg^-1). Except the control group,the rat models of peripheral nerve toxicity were established by intraperitomeal injection with oxaliplatin(4 mg·kg^-1);meanwhile,different doses of resveratrol were given by gavage. At the end of the experiment,rats were sacrificed,the relative expression levels of mRNA and protein of TRPV1 and TRPM8 in L4-5 dorsal root ganglion were detected by qPCR and Western Blot. Relative expression levels of protein kinase A(PKA),protein kinase Cε(PKCε)and proteinase-activated receptors 2(PAR2)were detected by Western Blot. Results Compared with control group,the mRNA and protein levels of TRPV1 and TRPM8 in L4-5 dorsal root ganglion of model group were significantly increased(P < 0.05, P < 0.01). Furthermore, the protein expression levels of p-PKA,p-PKCε and PAR2 in L4-5 dorsal root ganglion were significantly increased(P < 0.05,P < 0.01). Conversely, resveratrol not only significantly decreased the mRNA and protein expression levels of TRPV1 and TRPM8(P < 0.05,P < 0.01),but also remarkably inhibited the protein expression levels of p-PKA,p-PKCε and PAR2 in L4-5 dorsal root ganglion of rats(P < 0.05). Conclusion Resveratrol improves oxaliplatininduced peripheral nerve toxicity by decreasing the expression levels of TRPV1 and TRPM8 and inhibiting the activation of PAR2-PKA and PAR2-PKCε.