不同浓度丹参酮对大鼠脑片缺氧缺糖损伤的保护作用

Protective effects of tanshinone on brain slices injury induced by oxygen glucose deprivation

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。

AIM: To investigate the protective effects of tanshinone on the brain slices injury induced by oxygen glucose deprivation(OGD). METHODS: Rat brain slices were made and randomly assigned to four groups (n=10 in each):control group,OGD group,tanshinone 20 mg·L -1 group and tanshinone 200 mg·L -1 group. Changes of the neuron injury and apoptosis were observed with TTC staining, LDH releases, TUNEL staining, immunohistochemistry and electromicroscope. In addition, changes of intracellular calcium were measured by confocal laser-scanning microscopy. RESULTS: Tanshinone 20 mg·L -1 and 200 mg·L -1 attenuated the decrease of TTC staining and the increase of LDH release induced by OGD in brain slices. Neuronal apoptosis and changes of neuronal ultrastructures were attenuated by different concentrations of tanshinone. Bcl-2 and Bax protein expression was increased after OGD. Bax protein expression was decreased by different concentrations of tanshinone, while Bcl-2 protein expression was increased by different concentrations of tanshinone. Intracellular calcium concentration was increased by OGD and then it was lowered by different concentrations of tanshinone. The protective effects of 200 mg·L -1 tanshinone were more obvious better than those in the 20 mg·L -1 tanshinone group. CONCLUSION: Different concentrations of tanshinone have neuronprotective effects against OGD injury in rat brain slices, and 200 mg·L -1 tanshinone shows more obvious protective effects than those in the 20 mg·L -1 tanshinone.