Palo Titchaporn, Thaworn Athiwat, Charoenkij Phornnapa, Thamsermsang Onusa, Chotewuttakorn Sirikul, Tripatara Pinpat, Laohapand Tawee, Akarasereenont Pravit
Department of Pharmacology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.
Center of Applied Thai Traditional Medicine, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.
Evid Based Complement Alternat Med. 2017;2017:9383272. doi: 10.1155/2017/9383272. Epub 2017 Oct 29.
To investigate the modulated effects of HRF on cyclooxygenase isoform expression and its activity, using the human umbilical vein endothelial cell (HUVEC) model induced by interleukin-1 beta (IL-1).
Cells were treated with indomethacin (positive control), HRF, and its components at various concentrations prior to treatment with IL-1 at 24 h. Cell viability was determined by MTT assay. Moreover, the anti-inflammatory effects of HRF and its components through mRNA and protein expression were established using real-time quantitative PCR and Western blot, respectively. COX activity was identified via exogenous and endogenous PGE productions using the EIA.
There was no cytotoxicity in HUVECs treated with HRF. None of the experimental conditions used in the study affected the expression of COX-1, but COX-2 protein expression was inhibited at concentrations under 10 g/mL. Despite the significantly increased levels of exogenous PGE, HRF had no effect on COX-2 mRNA expression. However, the production of PGE was lower at a concentration of 100 g/mL HRF than at a concentration below 10 g/mL. Interestingly, each component of HRF revealed different effects of the Ha-Rak formula.
Our preliminary findings suggest that HRF and its components provide diverse modulation of COX-2 and PGE at the level.
利用白细胞介素-1β（IL-1）诱导的人脐静脉内皮细胞（HUVEC）模型，研究热射病方剂（HRF）对环氧合酶同工型表达及其活性的调节作用。
在24小时用IL-1处理之前，细胞用吲哚美辛（阳性对照）、HRF及其不同浓度的成分进行处理。通过MTT法测定细胞活力。此外，分别使用实时定量PCR和蛋白质印迹法，通过mRNA和蛋白质表达确定HRF及其成分的抗炎作用。通过酶免疫分析（EIA），利用外源性和内源性前列腺素E（PGE）的产生来鉴定COX活性。
用HRF处理的HUVECs没有细胞毒性。该研究中使用的任何实验条件均未影响COX-1的表达，但在浓度低于10μg/mL时，COX-2蛋白表达受到抑制。尽管外源性PGE水平显著升高，但HRF对COX-2 mRNA表达没有影响。然而，HRF浓度为100μg/mL时的PGE产生量低于浓度低于10μg/mL时。有趣的是，HRF的每种成分都显示出热射病方剂的不同作用。
我们的初步研究结果表明，HRF及其成分在水平上对COX-2和PGE具有多种调节作用。
