Jiang Enhui, Chen Xuanbo, Bi Yi, Pan Chuanying, Li Xiangchen, Lan Xianyong
Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China.
College of Animal Science and Technology, Zhejiang A&F University, Hangzhou 311300, China.
Animals (Basel). 2024 Nov 26;14(23):3421. doi: 10.3390/ani14233421.
In livestock production, oxidative stress (OS) is ubiquitous, reducing animal productivity and product quality. Hence, investigating the mechanisms of oxidative stress in livestock and inhibiting oxidative stress-induced damage is crucial. Curcumin, a plant-derived bioactive compound, exhibits antioxidant and anti-apoptotic properties. Adipose-derived stem cells (ADSCs) from animal adipose tissue are easily accessible and possess multilineage differentiation potential. Therefore, this work utilized bovine ADSCs to establish an oxidative stress model and investigated the effects of curcumin on oxidative stress and apoptosis. Firstly, bovine ADSCs were isolated and cultured from fetal calf subcutaneous adipose tissue. Their surface markers were identified by immunofluorescence, confirming the expression of CD29, CD44, CD73, CD90, CD105 and Vimentin, but not CD34, indicative of mesenchymal stem/progenitor cell characteristics. Secondly, to explore the effects of curcumin on oxidative damage and apoptosis in bovine ADSCs, an oxidative stress model was induced using HO. CCK-8 assays showed significantly reduced cell viability and SOD activity, along with increased malondialdehyde (MDA) and reactive oxygen species (ROS) levels, indicating successful modeling. RT-qPCR further confirmed that 500 μM of HO treatment for 24 h promoted apoptosis. Herein, CCK-8 assays indicated a significant reduction in cell viability at >8 μM of curcumin. Thirdly, using 4 μM and 8 μM of curcumin for pre-protection, 8 μM maintained SOD activity, reduced MDA and ROS, inhibited apoptosis-related gene changes (Bcl-2, Bax, Caspase-3), and suppressed apoptosis according to a TUNEL assay. Fourthly, curcumin's autophagy-inducing potential was hypothesized, which was confirmed by increased LC3-II and decreased P62 expression upon co-treatment with 3-MA. 3-MA inhibited curcumin's antioxidant and anti-apoptotic effects, suggesting that curcumin's antioxidant and anti-apoptotic roles may involve autophagy induction. In conclusion, bovine ADSCs are abundant, easily accessible, and multipotent, making them suitable for in vitro expansion. Curcumin alleviated HO-induced oxidative stress in bovine ADSCs, with curcumin also inhibiting apoptosis, likely through autophagy induction. This study validates the protective role of curcumin in bovine ADSCs, with potential applications in livestock production.
在畜牧生产中，氧化应激（OS）普遍存在，会降低动物的生产性能和产品质量。因此，研究家畜氧化应激的机制并抑制氧化应激引起的损伤至关重要。姜黄素是一种植物源生物活性化合物，具有抗氧化和抗凋亡特性。动物脂肪组织来源的脂肪干细胞（ADSCs）易于获取且具有多向分化潜能。因此，本研究利用牛ADSCs建立氧化应激模型，研究姜黄素对氧化应激和细胞凋亡的影响。首先，从胎牛皮下脂肪组织中分离培养牛ADSCs。通过免疫荧光鉴定其表面标志物，证实CD29、CD44、CD73、CD90、CD105和波形蛋白表达，但CD34不表达，表明具有间充质干/祖细胞特征。其次，为了探究姜黄素对牛ADSCs氧化损伤和细胞凋亡的影响，使用H₂O₂诱导建立氧化应激模型。CCK-8法检测显示细胞活力和超氧化物歧化酶（SOD）活性显著降低，丙二醛（MDA）和活性氧（ROS）水平升高，表明建模成功。RT-qPCR进一步证实500 μM H₂O₂处理24小时可促进细胞凋亡。在此，CCK-8法检测表明姜黄素浓度>8 μM时细胞活力显著降低。第三，分别用4 μM和8 μM姜黄素进行预处理，8 μM可维持SOD活性，降低MDA和ROS水平，抑制凋亡相关基因变化（Bcl-2、Bax、Caspase-3），并通过TUNEL检测抑制细胞凋亡。第四，推测姜黄素具有诱导自噬的潜能，与3-MA共同处理后LC3-II表达增加、P62表达降低证实了这一点。3-MA抑制了姜黄素的抗氧化和抗凋亡作用，表明姜黄素的抗氧化和抗凋亡作用可能涉及自噬诱导。总之，牛ADSCs数量丰富、易于获取且具有多能性，适合体外扩增。姜黄素减轻了H₂O₂诱导的牛ADSCs氧化应激，并抑制细胞凋亡，可能是通过诱导自噬实现的。本研究证实了姜黄素对牛ADSCs具有保护作用，在畜牧生产中具有潜在应用价值。
