Supplementary Material for: The Pharmacological Effects of Plant-Derived versus Synthetic Cannabidiol in Human Cell Lines

<b><i>Introduction:</i></b> Cannabidiol (CBD) can be isolated from <i>Cannabis sativa</i> L. or synthetically produced. The aim of this study was to compare the in vitro effects of purified natural and synthetic CBD to establish any pharmacological differences or superiority between sources. <b><i>Methods:</i></b> Six purified samples of CBD were obtained, 4 of these were natural and 2 synthetic. The anticancer effects of CBD were assessed in a human ovarian cancer cell line (SKOV-3 cells). The neuroprotective effects of CBD were assessed in human pericytes in a model of stroke (oxygen glucose deprivation [OGD]). The ability of CBD to restore inflammation-induced intestinal permeability was assessed in differentiated human Caco-2 cells (a model of enterocytes). <b><i>Results:</i></b> (1) In proliferating and confluent SKOV-3 cells, all CBD samples similarly reduced resazurin metabolism as a marker of cell viability in a concentration-dependent manner (<i>p</i> &lt; 0.001). (2) In pericytes exposed to OGD, all CBD samples similarly reduced cellular damage (measured by lactate dehydrogenase) at 24 h by 31–48% and reduced inflammation (measured by IL-6 secretion) by 30–53%. Attenuation of IL-6 was inhibited by 5HT_1A receptor antagonism for all CBD sources. (3) In differentiated Caco-2 cells exposed to inflammation (TNFα and IFNγ, 10 ng/mL for 24 h), each CBD sample increased the speed of recovery of epithelial permeability compared to control (<i>p</i> &lt; 0.05–0.001), which was inhibited by a CB₁ receptor antagonist. <b><i>Conclusion:</i></b> Our results suggest that there is no pharmacological difference in vitro in the antiproliferative, anti-inflammatory, or permeability effects of purified natural versus synthetic CBD. The purity and reliability of CBD samples, as well as the ultimate pharmaceutical preparation, should all be considered above the starting source of CBD in the development of new CBD medicines.

**引言：** 大麻二酚（Cannabidiol, CBD）可从大麻（Cannabis sativa L.）中提取，亦可通过合成方式制备。本研究旨在对比纯化后天然与合成型CBD的体外生物学效应，以明确两种来源间是否存在药理学差异或优劣性。
**方法：** 本研究共获取6份纯化CBD样品，其中4份为天然来源，2份为合成来源。采用人卵巢癌细胞系SKOV-3细胞，评估CBD的抗癌活性；采用氧糖剥夺（oxygen glucose deprivation, OGD）卒中模型中的人周细胞，评估CBD的神经保护作用；采用分化型人Caco-2细胞（肠上皮细胞模型），评估CBD对炎症诱导的肠道通透性损伤的修复能力。
**结果：** (1) 在增殖期及汇合期SKOV-3细胞中，所有CBD样品均以浓度依赖性方式降低作为细胞活力标志物的刃天青代谢活性（*p* < 0.001），且各组效应无显著差异。(2) 在经OGD处理的周细胞中，所有CBD样品均可在24小时时将细胞损伤（以乳酸脱氢酶释放量表征）降低31%~48%，并将炎症反应（以白细胞介素6（IL-6）分泌量表征）减轻30%~53%；且所有来源CBD的IL-6分泌抑制效应均可被5HT₁A受体拮抗剂阻断。(3) 在经炎症因子（肿瘤坏死因子α（TNFα）与干扰素γ（IFNγ），各10 ng/mL，作用24 h）处理的分化型Caco-2细胞中，相较于对照组，每份CBD样品均可加快上皮通透性的恢复速度（*p* < 0.05~0.001），且该效应可被CB₁受体拮抗剂阻断。
**结论：** 本研究结果表明，纯化后的天然与合成型CBD在体外的抗增殖、抗炎及肠道通透性修复效应上无药理学差异。在新型CBD药物的研发过程中，相较于CBD的初始来源，样品的纯度、可靠性以及最终的药物制剂形式均应作为更优先的考量因素。