Safaeian Leila, Shafiee Fatemeh, Haghighatnazar Samira
Department of Pharmacology and Toxicology, Isfahan Pharmaceutical Sciences Research Center, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran.
Department of Pharmaceutical Biotechnology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran.
Mol Biol Rep. 2023 Jan;50(1):389-397. doi: 10.1007/s11033-022-08042-4. Epub 2022 Nov 6.
Andrographolide (AG) is a lactone diterpene with valuable biological activities. This in vitro study evaluated whether AG can protect cardiomyocytes under toxicities triggered with anti-cancer chemotherapeutic agents, doxorubicin (DOX) and arsenic trioxide (ATO).
H9C2 cells were pretreated with AG (0.5-10 µM) for 24 h and then exposed to DOX (1 μM) or ATO (35 μM) for another 24 h period. For determination of cell viability or cytotoxicity, MTT and lactate dehydrogenase (LDH) assay were used. Total oxidant and antioxidant capacities were estimated by determining hydroperoxides and ferric reducing antioxidant power (FRAP) levels. Real time-polymerase chain reaction was also used for quantitative evaluation of TLR4 gene expression. AG inhibited cardiomyocytes proliferation at the concentrations of more than 20 μM. However, it considerably enhanced cell viability and decreased cytotoxicity of DOX and ATO at the concentration range of 2.5-10 μM in MTT and LDH assays. AG significantly declined hydroperoxides concentration in ATO-treated cardiomyocytes and raised FRAP value in DOX- and ATO-treated cells. Furthermore, AG notably lessened TLR4 expression in H9C2 cells after exposure to DOX- and ATO.
In conclusion, these data presented that AG was able to reverse DOX- and ATO-induced cardiotoxicity in vitro. The cardiomyocyte protective activities of AG may be due to the decrease in TLR4 expression and total oxidant capacity and increase in total antioxidant capacity.
穿心莲内酯（AG）是一种具有重要生物活性的二萜内酯。本体外研究评估了AG是否能在抗癌化疗药物阿霉素（DOX）和三氧化二砷（ATO）引发的毒性作用下保护心肌细胞。
用AG（0.5 - 10 μM）预处理H9C2细胞24小时，然后再用DOX（1 μM）或ATO（35 μM）处理24小时。使用MTT和乳酸脱氢酶（LDH）测定法来确定细胞活力或细胞毒性。通过测定氢过氧化物和铁还原抗氧化能力（FRAP）水平来估计总氧化剂和抗氧化剂能力。实时聚合酶链反应也用于TLR4基因表达的定量评估。AG在浓度超过20 μM时抑制心肌细胞增殖。然而，在MTT和LDH测定中，在2.5 - 10 μM的浓度范围内，它显著提高了细胞活力并降低了DOX和ATO的细胞毒性。AG显著降低了ATO处理的心肌细胞中的氢过氧化物浓度，并提高了DOX和ATO处理的细胞中的FRAP值。此外，AG显著降低了H9C2细胞在暴露于DOX和ATO后的TLR4表达。
总之，这些数据表明AG能够在体外逆转DOX和ATO诱导的心脏毒性。AG对心肌细胞的保护作用可能归因于TLR4表达的降低、总氧化剂能力的降低以及总抗氧化剂能力的增加。
