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目的:探讨不同浓度丹参酮对大鼠脑片缺氧缺糖损伤的保护作用及其机制。方法:建立大鼠脑片缺氧缺糖损伤模型,设立对照组、缺氧缺糖损伤组、丹参酮20mg·L-1组和丹参酮200mg·L-1组。利用2,3,5三苯基氯化四氮唑(TTC)染色定量比色、乳酸脱氢酶(LDH)、免疫组化、电镜评价不同浓度丹参酮对脑损伤的保护作用。同时激光共聚焦显微镜测定脑片胞内钙变化。结果:不同浓度丹参酮(20、200mg·L-1)抑制脑片OGD损伤所致的TTC染色降低,减少LDH释放,减轻神经元凋亡,改善神经元超微结构的病理损伤。OGD损伤增加bax和bcl2蛋白表达和胞内钙离子浓度。不同浓度丹参酮(20、200mg·L-1)进一步上调bcl2蛋白表达,降低bax蛋白表达,同时抑制胞内钙离子浓度。与丹参酮20mg·L-1相比,丹参酮200mg·L-1作用较强。结论:不同浓度丹参酮具有脑保护作用,能够抑制缺氧缺糖损伤导致的大鼠脑片神经元损伤及其凋亡过程,且丹参酮对于胞内钙离子的调控可能是其发挥保护作用的重要机制之一。
Objective: To investigate the protective effect and mechanism of different concentrations of tanshinone on oxygen-glucose deprivation injury in rat brain slices. METHODS: Rat models of hypoxia-glucose deprivation and hypoglycemia were established. The control group, hypoxic-hypoglycemia injury group, tanshinone 20 mg·L-1 group and tanshinone 200 mg·L-1 group were established. Using 2,3,5 triphenyltetrazolium chloride (TTC) staining quantitative colorimetric assay, lactate dehydrogenase (LDH), immunohistochemistry, and electron microscopy to evaluate the protective effect of different concentrations of tanshinone on brain injury. At the same time laser confocal microscopy determination of brain intracellular calcium changes. RESULTS: Different concentrations of tanshinone (20, 200 mg·L-1) inhibited the TTC staining caused by OGD injury in brain slices, reduced LDH release, reduced neuronal apoptosis, and improved neuron ultrastructural pathological injury. OGD injury increased bax and bcl2 protein expression and intracellular calcium ion concentration. Different concentrations of tanshinone (20, 200 mg·L-1) further up-regulated bcl2 protein expression, decreased bax protein expression, and inhibited intracellular calcium ion concentration. Compared with tanshinone 20 mg·L-1, tanshinone 200 mg·L-1 has a stronger effect. CONCLUSION: Different concentrations of tanshinone have brain protective effects and can inhibit the neuronal damage and apoptosis of rat brain slices caused by oxygen-glucose deprivation injury. The regulation of intracellular calcium by tanshinone may be an important mechanism for its protective effect. one.