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三七皂苷R_1(notoginsenoside R_1,NGR_1)是传统中药三七的重要活性成分,也是雌激素受体激动剂。因其蕴含抗炎、抗氧化、抗凋亡等多种功能,故在疾病治疗中被普遍运用。为阐明NGR_1在缺血缺氧性脑病(hypoxic-ischemic brain damage,HIBD)中的潜在神经保护作用机制,该研究采用原代皮层神经元建立氧糖剥夺再灌注(oxygen-glucose deprivation/reoxygenation,OGD/R)损伤模型,并且给予NGR_1及雌激素受体阻断剂ICI-182780处理,然后分别采用MTT,LDH和Hochest 33342染色检测神经元存活率、细胞膜完整性和凋亡,Western blot检测抗凋亡通路ATF6α,p-Akt,Akt蛋白及凋亡相关蛋白Bax,Cleaved Caspase-3的表达。结果显示:神经元在OGD/R损伤后,其细胞存活率明显下降(P<0.05)、细胞膜相对完整性显著降低(P<0.05)、细胞凋亡加重(P<0.05),ATF6α,p-Akt表达下调且促凋亡蛋白Bax,Cleaved Caspase-3表达增加(P<0.05)。NGR_1处理后能够减轻OGD/R造成的神经元细胞损伤,上调ATF6α表达、促进Akt磷酸化并且减少Bax,Cleaved Caspase-3蛋白的表达(P<0.05),但是,NGR_1的这些神经保护性作用能够被雌激素受体阻断剂ICI-182780阻断。研究结果证实:NGR_1在缺血缺氧情况下对神经元具有保护性作用,该保护作用可能是NGR_1通过雌激素受体调控ATF6/Akt信号通路实现的。
Notoginsenoside R_1 (NGR_1) is an important active ingredient of the traditional Chinese medicine Panax notoginseng and is also an estrogen receptor agonist. Because of its contains anti-inflammatory, anti-oxidation, anti-apoptosis and other functions, it is widely used in the treatment of diseases. In order to elucidate the potential neuroprotective mechanism of NGR_1 in hypoxic-ischemic brain damage (HIBD), we used primary cortical neurons to establish oxygen-glucose deprivation / reoxygenation (OGD) / R) injury model, and given NGR_1 and estrogen receptor blocker ICI-182780 treatment, and then used MTT, LDH and Hochest 33342 staining to detect neuronal survival, cell membrane integrity and apoptosis, Western blot detection of anti-apoptosis Expression of ATF6α, p-Akt, Akt and Apoptosis Associated Proteins Bax, Cleaved Caspase-3 in Peritoneal Cavity. The results showed that after OGD / R injury, the survival rate of neurons decreased significantly (P <0.05), the relative integrity of cell membrane decreased significantly (P <0.05), the apoptosis of cells increased (P <0.05) Akt expression was down-regulated and the expression of pro-apoptotic proteins Bax and Cleaved Caspase-3 were increased (P <0.05). NGR-1 treatment attenuated the neuronal damage induced by OGD / R, up-regulated the expression of ATF6α, promoted the phosphorylation of Akt and decreased the expression of Bax and Cleaved Caspase-3 protein (P <0.05). However, these neuroprotective effects of NGR_1 Blocked by estrogen receptor blocker ICI-182780. The results confirmed that NGR - 1 has a protective effect on neurons in the condition of hypoxia and hypoxia, and this protective effect may be that NGR - 1 regulates ATF6 / Akt signaling through the estrogen receptor.