Zeng Hai-Rong, Wang Bing, Zhao Zhen, Zhang Qi, Liang Mei-Yun, Yao Ya-Qi, Bian Ka, Zhang Wei-Rong
Experiment Center for Teaching and Learning, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Rood, Pudong New District, Shanghai 201203, China; Murad Research Center for Moderniszed Chinese Medicine, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Rood, Pudong New District, Shanghai 201203, China.
Experiment Center for Teaching and Learning, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Rood, Pudong New District, Shanghai 201203, China; School of Pharmacy, Shanghai University of Traditional Chinese Medicine, 1200, Cailun Road, Pudong New District, Shanghai 201203, China.
J Ethnopharmacol. 2016 Aug 2;189:132-8. doi: 10.1016/j.jep.2016.05.030. Epub 2016 May 16.
The Chinese herb compound prescription Viola yedoensis Makino Anti-itching Compound (VYAC), which consists of Viola yedoensis Makino, herb, Sophora flavescens Aiton, root, and Dictamnus dasycarpus Turcz, root and rhizome, has been traditionally used to treat various skin allergic inflammatory diseases in clinic.
The aim of this study is to investigate the effects of VYAC on degranulation and to determine its anti-inflammatory mechanism in RBL-2H3 mast cells.
VYAC was extracted with water-coction extraction (Shufen et al., 2012). The aqueous extracts were concentrated in vacuum under reduced pressure and lyophilized using a freeze dryer, and lyophilized powder was obtained. MTT was used to evaluate the cytotoxic of VYAC on RBL-2H3 cells. Degranulation was carried out with RBL-2H3 cell model, which was stimulated with A23187 plus PMA. β-Hexosaminidase and histamine were measured to evaluate degranulation. The mRNA levels of inflammation cytokines (IL-1β, TNF-α, IL-6, and iNOS) were investigated by RT-PCR to explain the anti-inflammatory mechanism of VYAC.
VYAC did not show cytotoxic effect on RBL-2H3 cells in the range of 25-400μg/mL. A higher dose of VYAC (800μg/mL) showed significant cytotoxicity (P<0.05). VYAC could significantly inhibit β-hexosaminidase and histamine release when treated with 100, 200, and 400μg/mL (P<0.05), but could not significantly inhibit β-Hexosaminidase and histamine release when treated with 25 and 50μg/mL (p>0.05). The mRNA levels of inflammatory cytokines (TNF-α, IL-1β, IL-6, and iNOS) could significantly decrease when treated with 200 and 400μg/mL (P<0.05) of VYAC, which were associated with the development of inflammation.
Results showed that VYAC inhibited β-hexosaminidase and histamine release, which was inhibit A23187 plus PMA stimulated RBL-2H3 cell degranulation and downregulated inflammatory cytokines (IL-1β, TNF-α, IL-6, and iNOS) expression to block inflammatory development.
中药复方紫花地丁止痒复方（VYAC）由紫花地丁、全草、苦参、根以及白鲜皮、根和根茎组成，在临床上传统用于治疗各种皮肤过敏性炎症疾病。
本研究旨在探讨VYAC对脱颗粒的影响，并确定其在RBL-2H3肥大细胞中的抗炎机制。
采用水煎煮法提取VYAC（舒芬等人，2012年）。水提取物在减压下真空浓缩，并用冷冻干燥机冻干，得到冻干粉。采用MTT法评价VYAC对RBL-2H3细胞的细胞毒性。用A23187加PMA刺激RBL-2H3细胞模型进行脱颗粒实验。检测β-己糖胺酶和组胺以评估脱颗粒情况。通过RT-PCR研究炎症细胞因子（IL-1β、TNF-α、IL-6和iNOS）的mRNA水平，以解释VYAC的抗炎机制。
在25-400μg/mL范围内，VYAC对RBL-2H3细胞未显示细胞毒性作用。较高剂量的VYAC（800μg/mL）显示出显著的细胞毒性（P<0.05）。当用100、200和400μg/mL处理时，VYAC能显著抑制β-己糖胺酶和组胺释放（P<0.05），但当用25和50μg/mL处理时，不能显著抑制β-己糖胺酶和组胺释放（P>0.05）。当用200和400μg/mL的VYAC处理时，炎症细胞因子（TNF-α、IL-1β、IL-6和iNOS）的mRNA水平可显著降低（P<0.05），这与炎症的发展有关。
结果表明，VYAC抑制β-己糖胺酶和组胺释放，即抑制A23187加PMA刺激的RBL-2H3细胞脱颗粒，并下调炎症细胞因子（IL-1β、TNF-α、IL-6和iNOS）表达以阻断炎症发展。
