桂枝拮抗麻黄中枢氧化应激损伤及NLRP3炎症小体上调作用分析

Effect of Cinnamomi Ramulus in Resisting Ephedrine Herba Central Oxidant Damages and NLRP3 Inflammasome Up-regulation

目的:研究桂枝对麻黄中枢神经系统损伤的保护作用,并初步探讨其保护作用机制。方法:KM小鼠按体重随机分为生理盐水组,麻黄组(10 g·kg^(-1)),麻黄-桂枝3∶2组(10 g·kg^(-1)+6.67 g·kg^(-1)),麻黄-桂枝3∶1组(10 g·kg^(-1)+3.33 g·kg^(-1)),桂枝组(6.67 g·kg^(-1)),灌胃给药14 d,酶联免疫吸附(ELISA)法检测各组小鼠脑组织中超氧化物歧化酶(SOD),丙二醛(MDA),一氧化氮(NO),一氧化氮合酶(NOS)活性,蛋白免疫印迹法(Western blot)测定小鼠脑组织NLRP3炎症小体表达水平。结果:与生理盐水组比较,麻黄组小鼠SOD活力下降(P<0.01),MDA,NO,TNOS,iNOS含量增加(P<0.01),Nod样受体蛋白3(NLRP3),凋亡相关斑点样蛋白(ASC)和半胱氨酸天冬氨酸蛋白酶-1(Caspase-1)蛋白表达上调(P<0.01),桂枝组无明显差异;与麻黄组比较,化学法结果显示麻黄-桂枝3∶2组和3∶1组均能够明显升高脑组织中SOD活力(P<0.05,P<0.01),降低MDA,NO,TNOS,iNOS含量(P<0.05,P<0.01),Western blot结果显示3∶2组能够显著降低NLRP3,ASC和Caspase-1蛋白表达(P<0.05,P<0.01),3∶1组能够降低Caspase-1蛋白表达(P<0.05)。结论:桂枝对麻黄中枢神经系统损伤具有保护作用,且具有一定的量效关系,随着桂枝比例的增加,桂枝拮抗麻黄中枢毒性更为显著,其作用机制可能与其增加抗氧化活性,减少氧化应激损伤,抑制NLRP3炎症小体上调有关。

Objective： To investigate the underlying mechanism of Cinnamomi Ramulus （CR）＇s protective effect on the central nervous system damages in mice induced by Ephedrine Herba （EH）. Method： According to weight, KM mice were randomly assigned into saline group, EH group, EH-CR 3 ： 2 group, EH-CR 3：1 group and CR group. After intragastric administration for 14 days, superoxide dismutase （SOD）, malondialdehyde （MDA）, nitric oxide （NO）, and nitric oxide synthase （NOS） levels in brain tissues were detected with ELISA, and NLRP3 inflammasome was detected by Western blot. Result： As compared with the normal saline group, SOD activity was reduced in EH group, while the levels of MDA, NO, TNOS and iNOS were increased （P 〈 0.01 ） ; the expression levels of Nod-like receptor protein 3 （NLRP3）, apoptosis-associated specklike protein （ASC） and cysteine aspartic acid protease-1 （ Caspase-1 ） were up-regulated （P 〈 0.01 ） ; but CR group showed no significant difference. As compared with EH group, CR combined with EH could significantly increase SOD activity in brain tissue （P 〈 0.05, P 〈 0.01 ）, reduce the MDA, NO, TNOS, iNOS production （P 〈 0.05, P 〈 0.01 ） , and down-regulate the NLRP3, ASC and Caspase-1 protein expressions （P 〈 0.05, P 〈 0.01）. Conclusion： CR has protective effect on the central nervous system damages induced by EH, and its mechanism may be associated with increasing antioxidant ability, reducing oxidant damage and attenuating NLRP3 inflammasome expression.