Kono Toru, Kaneko Atsushi, Matsumoto Chinami, Miyagi Chika, Ohbuchi Katsuya, Mizuhara Yasuharu, Miyano Kanako, Uezono Yasuhito
Hokkaido University, Sapporo, Japan Sapporo Higashi Tokushukai Hospital, Sapporo, Japan
Tsumura Research Laboratories, Tsumura and Co, Inashiki-gun, Ibaraki, Japan.
Integr Cancer Ther. 2014 Sep;13(5):435-45. doi: 10.1177/1534735413520035. Epub 2014 Feb 4.
Chemotherapy-induced oral mucositis (COM) is characterized by painful inflammation with prolonged damage that involves the pathological pain-evoking prostaglandin E2 (PGE2). We previously found that gargling with hangeshashinto (HST), a traditional Japanese medicine, was effective for the treatment of COM. However, little is known regarding the mechanisms. Our aim was to identify the active ingredients and clarify the characteristic effects of HST on the PGE2 system.
Prostanoids produced by human oral keratinocytes (HOK) stimulated with IL-1β were measured by enzyme immunoassay. Active ingredients that regulate PGE2 production were identified and quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and a culture system of HOK cells.
Inducible PGE2, PGD2, and PGF2α, metabolites of cyclooxygenase (COX) pathways, were reduced by HST (10-300 µg/mL) without inducing cytotoxicity. The active ingredients of HST were quantified by LC-MS/MS, and [6]-shogaol, [6]-gingerol, wogonin, baicalein, baicalin, and berberine were shown to reduce PGE2 production. A mixture of these 6 ingredients at concentrations equal to 300 µg/mL of HST strongly suppressed PGE2 production to the same level as HST. [6]-Shogaol and [6]-gingerol did not decrease COX-2 mRNA expression and mostly inhibited PGE2 metabolic activity in an assay using intact HOK cells, suggesting that they regulate PGE2 synthesis at the posttranscriptional level. Wogonin, baicalin, and berberine inhibited expression of COX-2 mRNA without affecting PGE2 metabolic activity. Moreover, wogonin, but not [6]-shogaol, suppressed phosphorylation of mitogen-activated protein kinases (p38s and JNKs).
These lines show that HST includes several PGE2-regulating ingredients that have different mechanisms and can function as a multicomponent and multitarget agent for treatment of COM, indicating that HST may be beneficial in a new medical strategy for COM treatment.
化疗诱导的口腔黏膜炎（COM）的特征是伴有长期损伤的疼痛性炎症，涉及可引发病理性疼痛的前列腺素E2（PGE2）。我们之前发现，用日本传统药物半夏泻心汤（HST）漱口对COM治疗有效。然而，其作用机制尚不清楚。我们的目的是确定其活性成分，并阐明HST对PGE2系统的独特作用。
通过酶免疫测定法测量白细胞介素-1β刺激的人口腔角质形成细胞（HOK）产生的前列腺素。通过液相色谱-串联质谱法（LC-MS/MS）和HOK细胞培养系统鉴定并定量调节PGE2产生的活性成分。
HST（10 - 300μg/mL）可降低环氧合酶（COX）途径的诱导型PGE2、PGD2和PGF2α代谢产物，且不诱导细胞毒性。通过LC-MS/MS对HST的活性成分进行定量，结果显示[6]-姜辣素、[6]-姜酚、汉黄芩素、黄芩苷、黄芩素和小檗碱可降低PGE2的产生。这6种成分以等于300μg/mL HST的浓度混合时，可强烈抑制PGE2的产生，使其达到与HST相同的水平。在使用完整HOK细胞的实验中，[6]-姜辣素和[6]-姜酚并未降低COX-2 mRNA表达，且主要抑制PGE2代谢活性，这表明它们在转录后水平调节PGE2合成。汉黄芩素、黄芩苷和小檗碱抑制COX-2 mRNA的表达，但不影响PGE2代谢活性。此外，汉黄芩素可抑制丝裂原活化蛋白激酶（p38和JNK）的磷酸化，而[6]-姜辣素则无此作用。
这些结果表明，HST含有多种调节PGE2的成分，其作用机制不同，可作为治疗COM的多组分、多靶点药物，这表明HST可能有利于COM治疗的新医学策略。
