雷公藤红素及其衍生物的体外神经保护作用与机制研究
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篇名: | 雷公藤红素及其衍生物的体外神经保护作用与机制研究 |
TITLE: | Neuroprotective effect and mechanism of celastrol and its derivatives in vitro |
摘要: | 目的 从神经炎症和氧化损伤2个方面探究雷公藤红素(Cel)及其衍生物Cel-1、Cel-2的神经保护作用及可能机制。方法利用1μg/mL脂多糖(LPS)诱导建立小胶质BV2细胞神经炎症模型、200μmol/L过氧化氢(H2O2)诱导建立人神经母细胞瘤SH-SY5Y细胞氧化损伤模型,考察不同浓度(0.625~20μmol/L)Cel、Cel-1、Cel-2对2种细胞的毒性;并在安全浓度(0.039~0.625μmol/L)范围内检测LPS诱导BV2细胞培养液中一氧化氮(NO)、肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、IL-6含量,检测H2O2诱导SH-SY5Y细胞的存活率。检测0.156、0.313、0.625μmol/L活性化合物Cel-2作用后H2O2诱导SH-SY5Y细胞中磷脂酰肌醇3激酶(PI3K)、磷酸化PI3K(p-PI3K)、蛋白激酶B(Akt)、磷酸化Akt(p-Akt)、胱天蛋白酶3(caspase-3)、B淋巴细胞瘤2(Bcl-2)和Bcl-2相关X蛋白(Bax)表达水平。结果在0.039~0.625μmol/L浓度梯度区间,神经炎症模型实验显示,Cel、Cel-1、Cel-2均可降低BV2细胞培养液中NO、TNF-α、IL-1β、IL-6的含量(P<0.05或P<0.01),其对神经炎症的半数抑制浓度分别为(0.25±0.04)、(0.61±0.14)、(0.11±0.02)μmol/L;氧化损伤模型实验显示,Cel、Cel-1、Cel-2在一定浓度下可逆转H2O2处理后SH-SY5Y细胞存活率降低的现象(P<0.05或P<0.01),其神经保护的半数效应浓度分别为(0.43±0.08)、(0.45±0.04)、(0.28±0.03)μmol/L。经H2O2诱导,Cel-2干预后SH-SY5Y细胞中PI3K、Akt蛋白的磷酸化水平和Bcl-2蛋白表达水平、Bcl-2/Bax比值均显著升高(P<0.05或P<0.01),caspase-3、Bax蛋白表达水平均显著降低(P<0.05或P<0.01)。结论Cel、Cel-1、Cel-2在一定浓度下均具有显著的神经保护活性,且以Cel-2的活性最强;Cel-2的作用机制可能与调控PI3K/Akt、caspase-3/Bcl-2/Bax信号通路,减轻炎症反应、氧化应激损伤,抑制神经细胞凋亡有关。 |
ABSTRACT: | OBJECTIVE To explore the neuroprotective effect and possible mechanism of celastrol (Cel) and its derivatives (Cel-1, Cel-2) in terms of neuroinflammation and oxidative damage. METHODS Neuroinflammation model of microglial BV2 cells was induced by 1 μg/mL lipopolysaccharide (LPS); oxidative damage model of human neuroblastoma SH-SY5Y cells was induced by 200 μmol/L hydrogen peroxide (H2O2). The toxicity of different concentrations of Cel, Cel-1 and Cel-2 (0.625-20 μmol/L) to the two types of cells was investigated. The levels of nitric oxide (NO), tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and IL-6 in BV2 cells induced by LPS at safe concentrations (0.039-0.625 μmol/L) were all detected. The survival rate of SH-SY5Y cells induced by H2O2 was also determined. The expression levels of phosphoinositide 3-kinase (PI3K), p-PI3K, protein kinase B (Akt), p-Akt, cystatinase 3 (caspase-3), B-cell lymphoma 2 (Bcl-2) and Bcl-2-related X protein (Bax) in SH- SY5Y cells induced by H2O2 at 0.156, 0.313, 0.625 μmol/L of active compound 2 were all detected. RESULTS In the concentration gradient range between 0.039 and 0.625 μmol/L, the results of neuroinflammation model experiments showed that Cel, Cel-1 and Cel-2 could reduce the contents of NO, TNF-α, IL-1β, and IL-6 in culture medium of BV2 cells (P<0.05 or P< 0.01); their IC50 values for neuroinflammation were (0.25±0.04), (0.61±0.14) and (0.11±0.02) μmol/L respectively. Meanwhile, all of them could reverse the phenomenon of decreased cell survival rate after H2O2 treatment in the oxidative damage experiments at a certain concentration (P< 0.05 or P<0.01), with neuroprotective EC50 values of (0.43± XJC2023009) 0.08), (0.45±0.04) and (0.28±0.03) μmol/L, respectively.Induced by H2O2, the phosphorylation of PI3K and Akt protein, protein expressions of Bcl-2 and Bcl-2/Bax ratio were all increased significantly (P<0.05 or P<0.01), while the protein expressions of caspase-3 and Bax were decreased significantly (P<0.05 or P<0.01). CONCLUSIONS Cel, Cel-1, and Cel-2 all have significant neuroprotective activities at certain concentrations, and Cel-2 shows the most significant protective effect. The mechanism of action of Cel-2 may be related to regulating the PI3K/Akt and caspase-3/Bcl-2/Bax signaling pathways, reducing the inflammatory response, oxidative stress damage and inhibiting neuronal apoptosis. |
期刊: | 2024年第35卷第05期 |
作者: | 陈佩佩;袁晓煊;张鑫;徐伟;许少华 |
AUTHORS: | CHEN Peipei,YUAN Xiaoxuan,ZHANG Xin,XU Wei,XU Shaohua |
关键字: | 雷公藤红素;衍生物;神经保护作用;神经炎症;氧化应激 |
KEYWORDS: | celastrol; derivatives; neuroprotective; neuroinflammation; oxidative stress |
阅读数: | 62 次 |
本月下载数: | 26 次 |
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