Olanlokun Oludele John, Oloke Kemi, Olorunsogo Olufunso Olabode
Laboratories for Biomembrane Research and Biotechnology, Department of Biochemistry, Faculty of Basic Medical Sciences, College of Medicine, University of Ibadan, Nigeria.
Avicenna J Phytomed. 2020 Mar-Apr;10(2):190-201.
Extracts of (AD) are used in folkloric medicine to treat several diseases and infections. However, their roles in mitochondrial permeability transition pore opening are not known.
The viability of mitochondria isolated from Wistar rat liver used in this experiment, was assessed by monitoring their swelling amplitude in the absence of calcium and reversal of calcium-induced pore opening by spermine. The effects of methanol extract and fraction of (MEAD and MFAD, respectively) on Mitochondrial Membrane Permeability Transition (MMPT) pore opening, ATPase activity, cytochrome c release and ferrous-induced lipid peroxidation were assessed spectrophotometrically. Phytochemical constituents of MEAD and MFAD were assessed using Gas Chromatography- Mass Spectrometry (GC-MS).
The MEAD (10, 20, 40 and 80 μg/ ml) had no effect on MMPT pore opening in the absence of Ca, whereas MFAD at 80 μg/ml had a large amplitude pore opening effect. Both MEAD and MFAD reversed Ca-induced swelling with inhibition values of 18, 21, 24, 23% (for MEAD) and 41, 36, 35, and 26% (for MFAD) at 10, 20, 40 and 80 μg/ml, respectively. MFAD significantly enhanced FF ATPase activity and caused cytochrome c release. Both MEAD and MFAD significantly inhibited ferrous-induced lipid peroxidation by 33.0, 64.0, 66, and 75% (for MEAD) and 24, 25, 30, and 45% (for MFAD), respectively. The GC-MS results revealed the presence of squalene as one of the major constituents of MEAD.
These findings suggest that MFAD can be used to induce cell death via mitochondrial permeability transition in isolated rat liver. Inhibition of lipid peroxidation by MEAD and MFAD showed that the pore opening effect of the extract and fraction was not mediated via peroxidation of mitochondrial membrane lipids.
（某物质，原文未明确写出具体名称）提取物在民间医学中用于治疗多种疾病和感染。然而，它们在线粒体通透性转换孔开放中的作用尚不清楚。
通过监测在无钙情况下线粒体的肿胀幅度以及精胺对钙诱导的孔开放的逆转情况，评估本实验中从Wistar大鼠肝脏分离的线粒体的活力。采用分光光度法评估甲醇提取物及其组分（分别为MEAD和MFAD）对线粒体膜通透性转换（MMPT）孔开放、ATP酶活性、细胞色素c释放和亚铁诱导的脂质过氧化的影响。使用气相色谱 - 质谱联用仪（GC - MS）评估MEAD和MFAD的植物化学成分。
在无钙情况下，MEAD（10、20、40和80μg/ml）对MMPT孔开放无影响，而80μg/ml的MFAD具有大幅度的孔开放效应。MEAD和MFAD均可逆转钙诱导的肿胀，在10、20、40和80μg/ml时，抑制率分别为18%、21%、24%、23%（MEAD）和41%、36%、35%、26%（MFAD）。MFAD显著增强FF ATP酶活性并导致细胞色素c释放。MEAD和MFAD均分别显著抑制亚铁诱导的脂质过氧化，抑制率为33.0%、64.0%、66%和75%（MEAD）以及24%、25%、30%和45%（MFAD）。GC - MS结果显示角鲨烯是MEAD的主要成分之一。
这些发现表明，MFAD可用于通过分离的大鼠肝脏中的线粒体通透性转换诱导细胞死亡。MEAD和MFAD对脂质过氧化的抑制表明提取物及其组分的孔开放效应不是通过线粒体膜脂质过氧化介导的。
