Department of Pharmacy, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200025, China.
Department of Pharmacy, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200025, China.
J Ethnopharmacol. 2020 Dec 5;263:113059. doi: 10.1016/j.jep.2020.113059. Epub 2020 Jul 11.
Shikonin, one of the main active ingredients of Chinese herbal medicine Lithospermum erythrorhizon, has been widely used to treat various disease including virus infection and inflammation in clinical. Its anti-tumor activity has been recorded in "Chinese herbal medicine". Recently, some studies about its anti-glioma effects have been reported. However, little is known about the molecular pharmacological activity of Shikonin in glioma.
This study aimed to systematically uncover and validate the pharmacological mechanism of Shikonin against glioma.
Network pharmacology approach, survival analysis, and Pearson co-expression analysis were performed to uncover and test the pharmacological mechanisms of Shikonin in glioma. Apoptosis assay, Caspase-3 activity assay and immunoblot analysis were practiced to validate the mechanisms.
Network pharmacology results suggested, anti-glioma effect of Shikonin by interfering endoplasmic reticulum (ER) stress-mediated tumor apoptosis targeting Caspase-3, and Bax/Bak-induced mitochondrial outer membrane permeabilization (MOMP) triggering cancer cell apoptosis. Survival analysis suggested the association of CASP3 with glioma (P < 0.05). Pearson correlation analysis indicated possible interaction of CASP3 with PERK through positive feedback regulation. Shikonin or in combination with 14G2a induced cell apoptosis in oligodendroglioma Hs683 cells in a dose-dependent manner with at a maximum apoptosis rate of 33%-37.5%, and 73%-77% respectively. Immunoblot analysis showed that Shikonin increased Caspase-3 activity to about 4.29 times, and increased 9 times when it combined with 14G2a. Shikonin increased also the expression levels of the proteins PERK and CHOP by about 4.4 and 5.6 folds, respectively, when it combined with 14G2a.
This study highlights the pharmacological mechanisms of Shikonin in the induction of tumor apoptosis in glioma cells.
紫草素是中药紫草中的主要活性成分之一，已广泛用于治疗各种疾病，包括病毒感染和炎症。其抗肿瘤活性已被《中药》所记载。最近，一些关于其抗神经胶质瘤作用的研究已经报道。然而，对于紫草素在神经胶质瘤中的分子药理学活性知之甚少。
本研究旨在系统地揭示和验证紫草素对神经胶质瘤的药理作用机制。
采用网络药理学方法、生存分析和 Pearson 共表达分析，揭示和验证紫草素治疗神经胶质瘤的药理机制。进行凋亡检测、Caspase-3 活性检测和免疫印迹分析以验证机制。
网络药理学结果表明，紫草素通过干扰内质网（ER）应激介导的肿瘤细胞凋亡靶向 Caspase-3，以及 Bax/Bak 诱导的线粒体膜通透性（MOMP）触发癌细胞凋亡，从而发挥抗神经胶质瘤作用。生存分析表明，CASP3 与神经胶质瘤有关（P<0.05）。Pearson 相关性分析表明，CASP3 可能通过正反馈调节与 PERK 相互作用。紫草素或与 14G2a 联合在寡树突胶质瘤 Hs683 细胞中呈剂量依赖性诱导细胞凋亡，最大凋亡率分别为 33%-37.5%和 73%-77%。免疫印迹分析显示，紫草素使 Caspase-3 活性增加约 4.29 倍，与 14G2a 联合使用时增加 9 倍。紫草素还使 PERK 和 CHOP 的蛋白表达水平分别增加约 4.4 倍和 5.6 倍，与 14G2a 联合使用时增加 9 倍。
本研究强调了紫草素在诱导神经胶质瘤细胞肿瘤细胞凋亡中的药理作用机制。
