Hou Qian-Ling, Wang Yan, Li Ying-Bo, Hu Xue-Lian, Wang Sha-Li
Institute of Neuroscience, School of Basic Medical Sciences, Chongqing Medical University, Chongqing 400016, China.
Department of Pharmacy, Second Affiliated Hospital, Third Military Medical University, Chongqing 400037, China.
Zhongguo Zhong Yao Za Zhi. 2017 Mar;42(6):1167-1174. doi: 10.19540/j.cnki.cjcmm.20170121.014.
Notoginsenoside R1(NGR1),a critical compound in traditional herb Panax notoginseng, is a kind of estrogen receptor agonist.It is reported to exhibit anti-apoptotic,anti-oxidative and anti-inflammatory properties activity, so it is widely used for treatment of various diseases.In order to investigate the potential neuroprotective effect of NGR1 in hypoxic-ischemic brain damage(HIBD), primary cortical neurons were used in this study to establish oxygen-glucose deprivation/reoxygenation(OGD/R) injury models. They were treated with NGR1 and estrogen receptor inhibitor ICI-182780 respectively, then the neuronal survival, cell membrane integrity and apoptosis were assessed by MTT assay,lactate dehydrogenase test(LDH) and Hoechst 33342 stain respectively, while the protein expression levels of ATF6α,p-Akt,Akt,Bax and Cleaved Caspase-3 were measured by Western blotting. Results indicated that as compared with the blank control group,OGD/R could induce cell injury and apoptosis(P<0.05), reduce relative integrity of cell membrane(P<0.05), decrease protein expression of ATF6α,p-Akt(P<0.05), and increase protein expression of Bax and Cleaved Caspase-3(P<0.05) in the primary cortical cells. After NGR1 treatment, the expression levels of ATF6α,p-Akt were obviously increased, and the expression levels of Bax and Cleaved Caspase-3 and the apoptosis of neuron were decreased(P<0.05). However, these neuroprotective properties of NGR1 against ODG/R-induced cell damage could be blocked by ICI-182780. This finding indicated that NGR1 may protect the primary cortical neurons against OGD/R induced injury,and the mechanism may be associated with accelerating the activation of the ATF6/Akt signaling pathway via estrogen receptors.
三七皂苷R1(NGR1)是传统草药三七中的一种关键化合物,是一种雌激素受体激动剂。据报道,它具有抗凋亡、抗氧化和抗炎活性,因此被广泛用于治疗各种疾病。为了研究NGR1在缺氧缺血性脑损伤(HIBD)中的潜在神经保护作用,本研究使用原代皮质神经元建立氧糖剥夺/复氧(OGD/R)损伤模型。分别用NGR1和雌激素受体抑制剂ICI-182780处理它们,然后分别通过MTT法、乳酸脱氢酶试验(LDH)和Hoechst 33342染色评估神经元存活、细胞膜完整性和凋亡情况,同时通过蛋白质印迹法检测ATF6α、p-Akt/Akt、Bax和裂解的半胱天冬酶-3的蛋白表达水平。结果表明,与空白对照组相比,OGD/R可诱导原代皮质细胞损伤和凋亡(P<0.05),降低细胞膜相对完整性(P<0.05),降低ATF6α、p-Akt的蛋白表达(P<0.05),并增加Bax和裂解的半胱天冬酶-3的蛋白表达(P<0.05)。NGR1处理后,ATF6α、p-Akt的表达水平明显升高,Bax和裂解的半胱天冬酶-3的表达水平及神经元凋亡减少(P<0.05)。然而,ICI-182780可阻断NGR1对OGD/R诱导的细胞损伤的这些神经保护特性。这一发现表明,NGR1可能保护原代皮质神经元免受OGD/R诱导的损伤,其机制可能与通过雌激素受体加速ATF6/Akt信号通路的激活有关。