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银杏二萜内酯葡胺注射液通过下调calpain信号通路抑制脑缺血神经细胞凋亡

Diterpene ginkgolides meglumine injection attenuates oxygen-glucose deprivation-induced apoptosis of nerve cells via inhibition of calpain signaling pathway

  • 摘要: 研究银杏二萜内酯葡胺注射液(diterpene ginkgolides meglumine injection,DGMI)对缺氧缺糖/复氧(oxygen-glucose deprivation/reoxygenation,OGD/R)损伤的人神经母细胞瘤细胞SH-SY5Y凋亡的抑制作用及其机制。采用试剂盒、Fluo-3 AM法、Western blot检测SH-SY5Y细胞氧糖剥离损伤4 h后,与药物一起复氧1 h细胞乳酸脱氢酶(LDH)的漏出量、caspase-3/7酶活力、细胞质中核小体含量、胞浆游离钙离子浓度以及calpain、cleaved caspase-12蛋白量的变化。结果显示DGMI能极显著降低OGD/R损伤的SH-SY5Y细胞LDH的漏出量,抑制caspase-3/7酶活性,减少细胞核核小体的释放量,下调细胞内游离钙离子浓度、calpain和cleaved caspase-12蛋白量,抑制calpain凋亡信号通路保护神经细胞。DGMI对OGD/R诱导的SH-SY5Y细胞凋亡具有显著的抑制作用,其作用机制可能与抑制细胞内Ca2+/calpain/caspase-12/capase-3信号通路有关。

     

    Abstract: To investigate the anti-apoptotic effect of diterpene ginkgolides meglumine injection(DGMI)on SH-SY5Y cells induced by oxygen-glucose deprivation/reoxygenation(OGD/R), and to explore its mechanisms. After 4 h of OGD, the SH-SY5Y cells were treated with 25 mg/L DGMI for 1 h. The release of lactic dehydrogenase(LDH)was measured by cytotoxicity detection kitplus. Cell apoptosis was detected by caspase-3/7 assays. Cell death was detected by ELISA. The concentration of [Ca2+i in cytoplasm was measured by Fluo-3 AM and the levels of calpain and cleaved capaease-12 were evaluated by western blot. As we expected, DGMI significantly decreased the release of LDH, the concentration of [Ca2+i, the protein levels of calpain and cleaved caspase-12. Furthermore, DGMI injection also attenuated the activities of caspase-3/7 and the contents of cytoplasmic histone-associated- DNA-fragments. These data demonstrated that the DGMI injection showed good anti-apoptotic effect in SH-SY5Y cells induced by OGD/R. The mechanisms may be associated with the inhibition of Ca2+/calpain/caspase-12/caspase-3 signaling pathway.

     

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