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α-细辛脑和β-细辛脑对 Aβ诱导的 PC12 细胞炎症反应的保护作用及其机制。

Protective effect of α-asarone and β-asarone on Aβ -induced inflammatory response in PC12 cells and its.

机构信息

3. School of Pharmacy, Hangzhou Medical College, Hangzhou 310053, China.

出版信息

Zhejiang Da Xue Xue Bao Yi Xue Ban. 2021 Oct 25;50(5):591-600. doi: 10.3724/zdxbyxb-2021-0162.

Abstract

To investigate effects of α-asarone and β-asarone on induced PC12 cell injury and related mechanisms. Aβ toxic injury cell model was induced by Aβ in PC12 cells. PC12 cells were divided into blank control group, model control group, α-asarone group (0.5, 1.0, β-asarone group (6.3, 12.5, vasoactive intestinal peptide (VIP) group, and VIP antagonist control group. Cell survival rate was detected by CCK-8 kit; cell apoptosis rate was detected by flow cytometry. The levels of inflammatory cytokines interleukin (IL)-1, , tumor necrosis factor (TNF)-α, oxidation-related inducible nitric oxide synthase (iNOS), nitric oxide (NO), apoptosis factors caspase-3 and p53 were detected by ELISA method. The expressions of C-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38MAPK) were detected by Western blotting. Compared with model control group, cell survival rates of group, β-asarone group and VIP group increased; the cell apoptosis rate decreased; levels of apoptosis-related factors caspase-3, p53, inflammatory factors IL-1, TNF-α decreased; IL-10 level increased; levels of oxidization-related factors iNOS and NO decreased; the expression of JNK and p38MAPK protein decreased (all <0.05). After VIP antagonist intervention, the survival rate of β-asarone group decreased; apoptosis rate increased; apoptosis related factors caspase-3, p53, inflammatory factors IL-1, TNF-α increased; IL-10 decreased; oxidation related factors iNOS and NO increased; the expression of JNK and p38MAPK protein increased (all <0.05); while there were no significant changes in these indicators of α-asarone group (all >0.05). α-asarone and β-asarone have protective effects on PC12 cell injury induced by Aβ. β-asarone may inhibit inflammatory factors and oxidation-related factors through promoting VIP secretion, regulating JNK/MAPK pathway, and reducing PC12 cell apoptosis; however, the effect of α-asarone may be not related to VIP secretion.

摘要

目的

探讨α-细辛脑和β-细辛脑对诱导的 PC12 细胞损伤的影响及其相关机制。

方法

将 PC12 细胞分为空白对照组、模型对照组、α-细辛脑组(0.5、1.0μmol/L)、β-细辛脑组(6.3、12.5μmol/L)、血管活性肠肽(VIP)组和 VIP 拮抗剂对照组。采用 CCK-8 试剂盒检测细胞存活率;采用流式细胞术检测细胞凋亡率;采用 ELISA 法检测白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α、氧化相关诱导型一氧化氮合酶(iNOS)、一氧化氮(NO)、凋亡因子 caspase-3 和 p53 的水平;采用 Western blot 法检测 c-Jun N-末端激酶(JNK)和 p38 丝裂原活化蛋白激酶(p38MAPK)的表达。

结果

与模型对照组比较,α-细辛脑组、β-细辛脑组和 VIP 组细胞存活率升高,细胞凋亡率降低,凋亡相关因子 caspase-3、p53,炎症因子 IL-1β、TNF-α水平降低,IL-10 水平升高,氧化相关因子 iNOS、NO 水平降低,JNK 和 p38MAPK 蛋白表达降低(均<0.05);给予 VIP 拮抗剂干预后,β-细辛脑组细胞存活率降低,细胞凋亡率升高,凋亡相关因子 caspase-3、p53,炎症因子 IL-1β、TNF-α水平升高,IL-10 水平降低,氧化相关因子 iNOS、NO 水平升高,JNK 和 p38MAPK 蛋白表达升高(均<0.05);而α-细辛脑组上述各指标均无明显变化(均>0.05)。

结论

α-细辛脑和β-细辛脑对 Aβ 诱导的 PC12 细胞损伤均具有保护作用,β-细辛脑可能通过促进 VIP 分泌,调节 JNK/MAPK 通路,减少氧化相关因子,降低 PC12 细胞凋亡,从而抑制炎症因子而发挥作用,而α-细辛脑的作用可能与 VIP 分泌无关。

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