Huang Chien-Hsun, Wang Fu-Ting, Chan Wen-Hsiung
Department of Obstetrics and Gynecology, Taoyuan General Hospital, Ministry of Health & Welfare, hongshan Road, Taoyuan District, Taoyuan City 33004, Taiwan.
Rehabilitation and Technical Aid Center, Taipei Veterans General Hospital, Shipai Road, Beitou District, Taipei City 11217, Taiwan.
Toxicol Res (Camb). 2020 Jul 1;9(4):431-443. doi: 10.1093/toxres/tfaa043. eCollection 2020 Jul.
Previous studies have shown that berberine, an isoquinoline alkaloid isolated from several traditional Chinese herbal medicines, suppresses growth and induces apoptosis in some tumor cell lines. It has also been shown that berberine possesses anti-atherosclerosis and antioxidant activities in hyperlipidemic model rats. Our previous study in mice found that berberine causes harmful effects on preimplantation and postimplantation embryonic development, both and , by triggering reactive oxygen species (ROS)-mediated apoptotic cascades in mouse blastocysts. In the current investigation, we further showed that berberine treatment has distinct dose-dependent effects on oocyte maturation and subsequent development. Preincubation of oocytes with 2.5 μM berberine significantly enhanced maturation and fertilization (IVF) rates, with subsequent beneficial effects on embryonic development. In contrast, preincubation with 10 μM berberine negatively impacted mouse oocyte maturation, decreased IVF rates and impaired subsequent embryonic development. Similar dose-dependent effects were also demonstrated . Specifically, intravenous injection of berberine significantly enhanced mouse oocyte maturation, IVF rate and early-stage embryo development after fertilization at a dose of 1 mg/kg body weight but significantly impaired oocyte maturation and IVF rates and caused harmful effects on early embryonic development at a dose of 5 mg/kg. Mechanistically, we found that berberine enhanced intracellular ROS production and apoptosis of oocytes at a concentration of 10 μM but actually significantly decreased total intracellular ROS content and had no apoptotic effect at a concentration of 2.5 μM. Moreover, pretreatment of oocytes with Ac-DEVD-cho, a caspase-3-specific inhibitor, effectively blocked berberine-induced negative impacts on oocyte maturation, fertilization and subsequent development. Collectively, these findings establish the dose-dependent beneficial versus deleterious effects of berberine and suggest that the mechanism underlying the deleterious effects of berberine involves a caspase-3-dependent apoptotic process acting downstream of an increase in intracellular ROS levels.
先前的研究表明，黄连素是从多种传统中草药中分离出的一种异喹啉生物碱，可抑制某些肿瘤细胞系的生长并诱导其凋亡。研究还表明，黄连素在高脂血症模型大鼠中具有抗动脉粥样硬化和抗氧化活性。我们之前在小鼠身上的研究发现，黄连素通过触发小鼠囊胚中活性氧（ROS）介导的凋亡级联反应，对植入前和植入后胚胎发育均产生有害影响。在当前的研究中，我们进一步表明，黄连素处理对卵母细胞成熟和随后的发育具有明显的剂量依赖性影响。用2.5 μM黄连素预孵育卵母细胞可显著提高成熟率和体外受精（IVF）率，并对胚胎发育产生后续有益影响。相比之下，用10 μM黄连素预孵育会对小鼠卵母细胞成熟产生负面影响，降低IVF率并损害随后的胚胎发育。在[具体实验或模型]中也证明了类似的剂量依赖性影响。具体而言，静脉注射黄连素在剂量为1 mg/kg体重时可显著提高小鼠卵母细胞成熟率、IVF率和受精后的早期胚胎发育，但在剂量为5 mg/kg时会显著损害卵母细胞成熟和IVF率，并对早期胚胎发育产生有害影响。从机制上讲，我们发现黄连素在浓度为10 μM时会增强卵母细胞内ROS的产生和凋亡，但在浓度为2.5 μM时实际上会显著降低细胞内总ROS含量，且无凋亡作用。此外，用caspase-3特异性抑制剂Ac-DEVD-cho预处理卵母细胞可有效阻断黄连素对卵母细胞成熟、受精和随后发育的负面影响。总的来说，这些发现确定了黄连素的剂量依赖性有益和有害作用，并表明黄连素有害作用的潜在机制涉及一个依赖caspase-3的凋亡过程，该过程在细胞内ROS水平升高的下游起作用。
