Department of Anesthesiology, Shanxian Central Hospital, Heze 274300, Shandong Province, China.
Department of Anesthesiology, Shanxian Central Hospital, Heze 274300, Shandong Province, China.
Life Sci. 2020 Oct 15;259:118279. doi: 10.1016/j.lfs.2020.118279. Epub 2020 Aug 13.
Bupivacaine, a common local anesthetic, can induce neurotoxicity and neurological complications. Capillarisin, a bioactive ingredient of Artemisia capillaris root extracts, has been reported to protect SH-SY5Y cells against oxidative stress-mediated neuronal cell death. Nevertheless, the effects of capillarisin on bupivacaine-induced neurotoxicity in SH-SY5Y cells remain unclear.
Cell viability, lactate dehydrogenase (LDH) release, reactive oxygen species (ROS) production, and apoptosis were detected. Malondialdehyde (MDA) content, glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) activities were measured for evaluation of oxidative stress. Western blot was performed to detect the changes of phosphatidylinositol-3-kinase (PI3K)/protein kinase B (PKB) pathway, and expression of cleaved poly ADP ribose polymerase (PARP), cleaved caspase-3, glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP). Activities of mitochondrial respiratory chain complexes I-III and adenosine triphosphate (ATP) content were measured to evaluate mitochondrial damage.
Bupivacaine treatment dose-dependently reduced cell viability, increased LDH release, and induced ROS production and PI3K/PKB pathway inactivation in SH-SY5Y cells, which were overturned by capillarisin treatment. Capillarisin inhibited bupivacaine-induced apoptosis in SH-SY5Y cells by decreasing cleaved PARP and cleaved caspase-3 expression. Capillarisin inhibited bupivacaine-induced oxidative stress, decrease of mitochondrial respiratory chain complex I, II, and III activities and ATP content, and increase of GRP78 and CHOP expression in SH-SY5Y cells. However, treatment with LY294002 abolished the effects of capillarisin on bupivacaine-induced neurotoxicity in SH-SY5Y cells.
Capillarisin protected SH-SY5Y cells against bupivacaine-induced apoptosis by inhibiting oxidative stress, mitochondrial injury, and endoplasmic reticulum stress via ROS-mediated of PI3K/PKB pathway.
布比卡因是一种常用的局部麻醉剂，可诱导神经毒性和神经并发症。青蒿素是青蒿根提取物的一种生物活性成分，已被报道可保护 SH-SY5Y 细胞免受氧化应激诱导的神经元细胞死亡。然而，青蒿素对 SH-SY5Y 细胞中布比卡因诱导的神经毒性的影响尚不清楚。
检测细胞活力、乳酸脱氢酶（LDH）释放、活性氧（ROS）产生和细胞凋亡。通过测定丙二醛（MDA）含量、谷胱甘肽过氧化物酶（GSH-Px）、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活性来评估氧化应激。通过 Western blot 检测磷脂酰肌醇-3-激酶（PI3K）/蛋白激酶 B（PKB）通路的变化，以及裂解多聚 ADP 核糖聚合酶（PARP）、裂解半胱天冬酶-3、葡萄糖调节蛋白 78（GRP78）和 C/EBP 同源蛋白（CHOP）的表达。测定线粒体呼吸链复合物 I-III 的活性和三磷酸腺苷（ATP）含量，以评估线粒体损伤。
布比卡因处理呈剂量依赖性降低 SH-SY5Y 细胞活力，增加 LDH 释放，并诱导 ROS 产生和 PI3K/PKB 通路失活，青蒿素处理可逆转上述作用。青蒿素通过降低裂解 PARP 和裂解半胱天冬酶-3 的表达抑制 SH-SY5Y 细胞中布比卡因诱导的细胞凋亡。青蒿素抑制布比卡因诱导的 SH-SY5Y 细胞氧化应激、线粒体呼吸链复合物 I、II 和 III 活性和 ATP 含量降低以及 GRP78 和 CHOP 表达增加。然而，LY294002 处理可消除青蒿素对 SH-SY5Y 细胞中布比卡因诱导的神经毒性的作用。
青蒿素通过 ROS 介导的 PI3K/PKB 通路抑制氧化应激、线粒体损伤和内质网应激，保护 SH-SY5Y 细胞免受布比卡因诱导的细胞凋亡。
