中国中药杂志

2016, v.41(09) 1627-1634

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基于致炎毒性,胃和十二指肠AQP3,AQP4表达的藤黄炮制减毒机制研究
Attenuation mechanism of gamboges during processing based on inflammatory toxicity and AQP3,AQP4 protein and m RNA expressions in rat gastric and duodenal tissues

赵琴;房芸;潘凌云;徐敏;王海颖;修彦凤;
ZHAO Qin;FANG Yun;PAN Ling-yun;XU Min;WANG Hai-ying;XIU Yan-feng;School of Pharmacy,Shanghai University of Traditional Chinese Medicine;Shanghai Fengxian District Hospital of Traditional Chinese Medicine;

摘要(Abstract):

为了拓展藤黄在中医临床的应用,研究其内服的毒性及炮制减毒的机制是必要的。通过巨噬细胞RAW264.7释放炎症介质(一氧化氮NO、肿瘤坏死因子TNF-α和白细胞介素IL-6)和灌胃给予藤黄生品和炮制品后大鼠胃和十二指肠组织的病理表现,判断其毒性作用;采用免疫组化和实时荧光定量PCR技术检测灌胃给药后,大鼠胃和十二指肠组织AQP3,AQP4蛋白和m RNA的表达,研究藤黄炮制减毒的机制。结果表明,藤黄生品可促进炎症介质NO,TNF-α和IL-6的释放,且与剂量呈相关性;藤黄制品组与生品组比较,NO和IL-6的释放量降低,TNF-α的释放量增加;藤黄生品可引起大鼠腹泻、白细胞升高、淋巴细胞降低,使胃黏膜充血水肿,肠黏膜坏死和炎细胞浸润,从多个角度证明内服生藤黄对胃和十二指肠组织的毒性为致炎毒性,致炎毒性与给药剂量呈相关性,炮制后藤黄的致炎毒性降低。在藤黄对胃和十二指肠组织致炎的同时,藤黄生品高剂量组大鼠胃和十二指肠组织水通道蛋白AQP3,AQP4 m RNA和蛋白表达量显著增加(P<0.05),相应剂量藤黄制品组大鼠AQP3,AQP4表达量较生藤黄组低,说明AQP3,AQP4蛋白和m RNA表达量的高低与藤黄的致炎作用强弱有一致性。通过降低AQP3,AQP4的表达水平可能是藤黄炮制减毒的作用机制之一。
To expand the clinical application of gamboges,it is necessary to study crude gamboges' toxicity after oral administration and attenuation mechanism during processing. In this study,crude gamboges' toxicity was judged by multiple assays,including inflammatory mediums [such as nitric oxide( NO),tumor necrosis factor alpha( TNF-α),and interleukin 6( IL-6) ] released by macrophage RAW264. 7,and pathological manifestations of rat stomach and duodenal tissues after oral administration with crude and processed gamboges. The attenuation mechanism during processing was studied by detecting AQP3,AQP4 protein and m RNA expression in rat gastric and duodenal tissues using immunohistochemical assay and real-time fluorescent quantitative PCR technique. According to the results,crude gamboges group showed promotion in release of NO,TNF-α and IL-6 by macrophage RAW264. 7 in a dose-dependent manner; Compared with crude gamboges group,processed gamboges group showed reduction in release of NO and IL-6,with increase in TNF-α. Crude gamboges could cause rat diarrhea,white blood cells increase,lymphocytes decrease,hyperemia and edema in rat gastric mucosa,duodenal mucosal necrosis and inflammatory cells infiltration. All of these results proved that gamboges had the inflammatory toxicity in gastric and duodenal tissues after oral administration in a dose-dependent manner,which however reduced after processing. In addition to the inflammatory toxicity,the m RNA and protein expressions of aquaporin 3( AQP3),aquaporin 4( AQP4)in gastric and duodenal tissues of high-dose crude gamboges group were increased significantly( P < 0. 05),while the protein and m RNA expressions of AQP3,AQP4 were weakened in processed gamboges group. The results showed that AQP3,AQP4 protein and m RNA expressions were positively correlated with the inflammatory toxicity. In conclusion,down-regulation of AQP3,AQP4 protein and m RNA expressions may be one of attenuation mechanisms in processing gamboges.

关键词(KeyWords): 藤黄;炮制;毒性;致炎;AQP3;AQP4
gamboges;processing;toxicity;inflammation;aquaporin 3(AQP3);aquaporin 4(AQP4)

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基金项目(Foundation): 上海市自然科学基金项目(13ZR1442000);; 上海市教委预算内课题项目(2014YSN20)

作者(Author): 赵琴;房芸;潘凌云;徐敏;王海颖;修彦凤;
ZHAO Qin;FANG Yun;PAN Ling-yun;XU Min;WANG Hai-ying;XIU Yan-feng;School of Pharmacy,Shanghai University of Traditional Chinese Medicine;Shanghai Fengxian District Hospital of Traditional Chinese Medicine;

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