Liu Juxiang, Zhang Lianli, Liu Dan, Li Baocai, Zhang Mi
Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, China.
Comb Chem High Throughput Screen. 2018;21(8):571-582. doi: 10.2174/1386207321666181005121457.
Aim & Objectives: Curcuminoids are characteristic constituents in Curcuma, displaying obviously neuroprotective activities against oxidative stress. As one of the Traditional Chinese Medicines from Curcuma, the radix of Curcuma aromatica is also rich in those chemicals, but its neuroprotective activity and mechanism remain unknown. The aim of the current study is to evaluate the neuroprotective effects of extracts from the radix of C. aromatica (ECAs) on H2O2-damaged PC12 cells.
The model of oxidative stress damage was established by treatment of 400 µM H2O2 on PC12 to induce cell damage. After the treatment of ECWs for 24 h, the cell viability, LDH, SOD, CAT and GSH were measured to evaluate the neuroprotection of ECAs on that model. The potential action mechanism was studied by measurement of level of ROS, cell apoptosis rate, mitochondrial membrane potential (MMP), morphologic change, the intracellular Ca2+ content (F340/F380) and the expressions of Bcl-2, Bax and Caspase-3. Additionally, the constituents from tested extracts were analyzed by HPLC-DAD-Q-TOF-MS method.
Compared with a positive control, Vitamin E, 10 µg/ml of 95% EtOH extract (HCECA) and 75% EtOH extract (MCECA) can markedly increase the rate of cell survival and enhance the antioxidant enzyme activities of SOD, CAT, increase the levels of GSH, decrease LDH release and the level of ROS, attenuate the intracellular Ca2+ overloading, reduce the cell apoptotic rate and stabilize MMP, down-regulate Bcl-2 expression, up-regulate Bax and caspase-3 expression, and improve the change of cell morphology. The chemical analysis showed that diarylheptanoids and sesquiterpenoids are the major chemicals in tested extracts and the former were richer in HCECA and MCECA than others.
These findings indicated that the effects of HCECA and MCECA on inhibiting the cells damage induced by H2O2 in PC12 are better than other extracts from the radix of C. aromatica, and the active constituents with neuroprotective effects consisting in those two active extracts are diarylheptanoids.
目的与目标：姜黄素类化合物是姜黄中的特征性成分，对氧化应激具有明显的神经保护活性。作为源自姜黄的传统中药之一，郁金的根也富含这些化学成分，但其神经保护活性及机制仍不清楚。本研究旨在评估郁金根提取物（ECAs）对H2O2损伤的PC12细胞的神经保护作用。
通过用400 μM H2O2处理PC12细胞以诱导细胞损伤，建立氧化应激损伤模型。在用ECWs处理24小时后，测量细胞活力、乳酸脱氢酶（LDH）、超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和谷胱甘肽（GSH），以评估ECAs对该模型的神经保护作用。通过测量活性氧（ROS）水平、细胞凋亡率、线粒体膜电位（MMP）、形态变化、细胞内钙离子含量（F340/F380）以及Bcl-2、Bax和Caspase-3的表达，研究其潜在作用机制。此外，采用高效液相色谱-二极管阵列-四极杆-飞行时间质谱（HPLC-DAD-Q-TOF-MS）法分析受试提取物中的成分。
与阳性对照维生素E相比，10 μg/ml的95%乙醇提取物（HCECA）和75%乙醇提取物（MCECA）可显著提高细胞存活率，增强SOD、CAT的抗氧化酶活性，增加GSH水平，降低LDH释放和ROS水平，减轻细胞内钙离子超载，降低细胞凋亡率并稳定MMP，下调Bcl-2表达，上调Bax和caspase-3表达，并改善细胞形态变化。化学分析表明，二芳基庚烷类化合物和倍半萜类化合物是受试提取物中的主要化学成分，且HCECA和MCECA中的前者比其他提取物中的含量更丰富。
这些发现表明，HCECA和MCECA对抑制PC12细胞中H2O2诱导的细胞损伤的作用优于郁金根的其他提取物，且这两种活性提取物中具有神经保护作用的活性成分是二芳基庚烷类化合物。
