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

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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 study the protective effect of Guizhi on the central nervous system injury of Ephedra and to explore its protective mechanism. Methods KM mice were randomly divided into normal saline group, ephedrine group (10 g · kg -1), Ephedra sinicola 3: 2 group (10 g · kg -1) kg -1 in the group of Ephedra sinica and Guizhi 3:1 group (10 g · kg -1 and +3.33 g · kg -1), while those in the Guizhi group (6.67 g · kg -1 )) Were administered intragastrically for 14 days. The contents of superoxide dismutase (SOD), malondialdehyde (MDA), nitric oxide (NO), monoxide Nitrogen synthase (NOS) activity and Western blot were used to detect the expression of NLRP3 inflammasome in mouse brain. Results: Compared with the saline group, the activity of SOD in ephedra decreased (P <0.01), the contents of MDA, NO, TNOS and iNOS increased (P <0.01), Nod like receptor 3 (NLRP3) The expression of ASC and Caspase-1 was up-regulated (P <0.01), but there was no significant difference in Guizhi group. Compared with the control group, The levels of MDA, NO, TNOS and iNOS in the 3: 2 and 3: 1 groups were significantly increased (P <0.05, P <0.01) The results of blot showed that 3: 2 group could significantly reduce the expression of NLRP3, ASC and Caspase-1 protein (P <0.05, P <0.01), and the 3: 1 group could reduce the expression of Caspase-1 protein. Conclusion: Guizhi has a protective effect on central nervous system injury of ephedra and has a certain dose-effect relationship. With the increase of Guizhi ratio, the Guizhi antagonistic ephedra central toxicity is more significant, its mechanism may be related to its anti-oxidation activity, Reduce oxidative stress injury, inhibition of NLRP3 inflammasome upregulation.
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