Pharmacen™, Centre of Excellence for Pharmaceutical Sciences, North-West University, Private Bag X6001, Potchefstroom, 2520, South Africa.
Department of Biochemistry and Molecular Biology, University of Southern Denmark, DK-5230, Odense M, Denmark; Celvivo IVS, Blommenslyst, Denmark.
J Ethnopharmacol. 2019 Jul 15;239:111897. doi: 10.1016/j.jep.2019.111897. Epub 2019 Apr 19.
Traditional herbal medicines are utilized by 27 million South Africans. Xysmalobium undulatum (Uzara) is one of the most widely used traditional medicinal plants in Southern Africa. A false belief in the safety of herbal medicine may result in liver injury. Herb-induced liver injury (HILI) range from asymptomatic elevation of liver enzymes, to cirrhosis and in certain instances even acute liver failure. Various in vitro and in vivo models are available for the pre-clinical assessment of drug and herbal hepatotoxicity. However, more reliable and readily available in vitro models are needed, which are capable of bridging the gap between existing models and real human exposure. Three-dimensional (3D) spheroid cultures offer higher physiological relevance, overcoming many of the shortcomings of traditional two-dimensional cell cultures.
This study investigated the hepatotoxic and anti-prolific effects of the crude X. undulatum aqueous extract during a sub-chronic study (21 days), in both a 3D HepG2/C3A spheroid model and the Sprague Dawley rat model.
HepG2/C3A spheroids were treated with a known hepatotoxin, valproic acid, and crude X. undulatum aqueous extract for 21 days with continuous evaluation of cell viability and proliferation. This was done by evaluating cell spheroid growth, intracellular adenosine triphosphate (ATP) levels and extracellular adenylate kinase (AK). Sprague Dawley rats were treated with the same compounds over 21 days, with evaluation of in vivo toxicity effects on serum chemistry.
The results from the in vitro study clearly indicated hepatotoxic effects and possible liver damage following treatment with valproic acid, with associated growth inhibition, loss of cell viability and increased cytotoxicity as indicated by reduced intracellular ATP levels and increased AK levels. These results were supported by the increased in vivo levels of AST, ALT and LDH following treatment of the Sprague Dawley rats with valproic acid, indicative of hepatic cellular damage that may result in hepatotoxicity. The in vitro 3D spheroid model was also able to predict the potential concentration dependant hepatotoxicity of the crude X. undulatum aqueous extract. Similarly, the results obtained from the in vivo Sprague Dawley model indicated moderate hepatotoxic potential.
The data from both the 3D spheroid model and the Sprague Dawley model were able to indicate the potential concentration dependant hepatotoxicity of the crude X. undulatum aqueous extract. The results obtained from this study also confirmed the ability of the 3D spheroid model to effectively and reliably predict the long-term outcomes of possible hepatotoxicity.
有 2700 万南非人使用传统草药。Xysmalobium undulatum（Uzara）是南非最广泛使用的传统药用植物之一。对草药安全性的错误信念可能导致肝损伤。草药引起的肝损伤（HILI）范围从无症状的肝酶升高，到肝硬化，在某些情况下甚至急性肝衰竭。有各种体外和体内模型可用于临床前评估药物和草药的肝毒性。然而，需要更可靠和易于获得的体外模型，这些模型能够弥合现有模型与真实人体暴露之间的差距。三维（3D）球体培养提供了更高的生理相关性，克服了传统二维细胞培养的许多缺点。
本研究在为期 21 天的亚慢性研究中，研究了粗 X. undulatum 水提取物的肝毒性和抗增殖作用，包括 3D HepG2/C3A 球体模型和 Sprague Dawley 大鼠模型。
用已知的肝毒素丙戊酸和粗 X. undulatum 水提取物处理 HepG2/C3A 球体 21 天，并连续评估细胞活力和增殖。通过评估细胞球体生长、细胞内三磷酸腺苷（ATP）水平和细胞外腺苷酸激酶（AK）来实现。用相同的化合物处理 Sprague Dawley 大鼠 21 天，评估血清化学的体内毒性作用。
体外研究结果清楚地表明，在用丙戊酸处理后，细胞毒性作用和可能的肝损伤，与生长抑制、细胞活力丧失和细胞毒性增加有关，这表现为细胞内 ATP 水平降低和 AK 水平升高。这些结果得到了 Sprague Dawley 大鼠用丙戊酸治疗后 AST、ALT 和 LDH 水平升高的支持，表明可能导致肝毒性的肝细胞损伤。体外 3D 球体模型还能够预测粗 X. undulatum 水提取物的潜在浓度依赖性肝毒性。同样，从 Sprague Dawley 大鼠体内模型获得的结果表明存在中度肝毒性潜力。
3D 球体模型和 Sprague Dawley 模型的数据均能够表明粗 X. undulatum 水提取物的潜在浓度依赖性肝毒性。本研究的结果还证实了 3D 球体模型能够有效可靠地预测可能肝毒性的长期结果。
