铁皮石斛不同部位对辣椒水复合氨水刺激致慢性咽炎模型大鼠的作用研究  

Effects of different extracts of Dendrobium officinale on rats with chronic pharyngitis induced by pepper water combined with ammonia

作　　者：笈文娜 颜美秋[1] 苏洁[1] 俞静静[1] 陈素红[2] 吕圭源[1] 陈建真[1] JI Wen-na;YAN Mei-qiu;SU Jie;YU Jing-ing;CHEN Su-hong;LYU Gui-yuan;CHEN Jian-zhen(Zhejiang Chinese Medical Universiy,Hangzhou 310053,China;Zhejiang Unirersity of Technology,Hangzhou 310014,China)

机构地区：[1]浙江中医药大学,浙江杭州310053 [2]浙江工业大学,浙江杭州310014

出　　处：《中国中药杂志》2022年第9期2525-2532,共8页China Journal of Chinese Materia Medica

基　　金：国家重点研发计划项目(2017YFC1702203);国家自然科学基金项目(81703772);浙江省中医药科技计划项目(2022ZB095);浙江省省级重点实验室项目(2012E10002)。

摘　　要：铁皮石斛“滋阴清热”“生津”,民间认为其能“清咽利喉”,但物质基础不清。为明确铁皮石斛对慢性咽炎的具体药效部位和潜在机制,该研究采用辣椒水复合低浓度氨水刺激法建立大鼠慢性咽炎模型,连续6周灌胃给予铁皮石斛茎粗多糖、非多糖部位及总提取物(0.33 g·kg^(-1),按生药计)。观测慢性咽炎大鼠口腔分泌物量、咽部表观状态变化,采用XT-2000i全自动血液分析仪测定血液学指标,ELISA法检测血清促炎因子肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、白细胞介素6(IL-6)和T淋巴细胞因子干扰素γ(IFN-γ)、白细胞介素4(IL-4)、白细胞介素17(IL-17)、转化生长因子β1(TGF-β1)水平,流式细胞仪测定外周血T淋巴细胞亚群CD3^(+)、CD4^(+)、CD8^(+)细胞比例,通过苏木素-伊红染色法观察咽部组织形态学变化,免疫组化法和蛋白质免疫印迹法检测咽部核因子-κB P65(NF-κB P65)、环氧化酶-2(COX-2)、F4/80、单核细胞趋化因子-1(MCP-1)的蛋白表达。结果显示,铁皮石斛粗多糖部位和非多糖部位均能显著减轻咽部病变,降低白细胞(WBC)、淋巴细胞(LYMP)数目以及促炎因子TNF-α、IL-6及IL-1β水平,抑制咽部NF-κB P65、COX-2、F4/80、MCP-1的蛋白表达;粗多糖部位降低口腔分泌物量和血清IL-17水平作用优于非多糖部位;非多糖部位增加CD4^(+)/CD8^(+)比值的作用优于粗多糖。提示铁皮石斛粗多糖和非多糖部位均有抗慢性咽炎作用,其抗炎机制可能与调节T淋巴细胞分布,抑制NF-κB P65介导的炎症信号通路有关;粗多糖部位抗炎作用可能与调节Th17/Treg平衡有关,非多糖部位则可能与调节Th、Tc比例有关。Dendrobium officinale can serve as Chinese medicinal material effective in nourishing yin,clearing heat,and producing fluid,and is used to treat throat diseases,but its active substances and mechanism are not clear.To clarify the active fraction and underlying mechanism of D.officinale against chronic pharyngitis(CP),the present study induced a CP model in rats by pepper water combined with low-concentration ammonia,and crude polysaccharides of D.officinale(DOP),non-polysaccharides of D.officinale(DON),and total extract of D.officinale(DOT)(0.33 g·kg^(-1),calculated according to the crude drug)were administered by gavage for six weeks.The changes in oral secretions and pharyngeal conditions of rats with CP were observed and rated.The hematological indicators were determined by an automatic hematology analyzer.The serum levels of pro-inflammatory factors,such as tumor necrosis factor-alpha(TNF-α),interleukin 1β(IL-1β),and interleukin 6(IL-6),and T-lymphocyte cytokines,including interferonγ(IFN-γ),interleukin 4(IL-4),interleukin 17(IL-17),and transforming growth factorβ1(TGF-β1)were detected by the enzyme-linked immunosorbent assay(ELISA).The proportions of CD3^(+),CD4^(+),and CD8^(+)cells in peripheral blood T lymphocyte subsets were determined by the flow cytometry.The histomorphological changes of the pharynx were observed by hematoxylin-eosin(HE)staining.The protein expression of nuclear factor-κB P65(NF-κB P65),cyclooxygenase-2(COX-2),F4/80,and monocyte chemoattractant protein-1(MCP-1)in the pharynx were detected by immunohistochemistry and Western blot.The results showed that DOP and DON could significantly relieve pharyngeal lesions,reduce white blood cells(WBC)and lymphocytes(LYMP),decrease the levels of pro-inflammatory factors TNF-α,IL-6,and IL-1β,and inhibit the protein expression of NF-κB P65,COX-2,F4/80,and MCP-1 in the pharynx.DOP was superior in reducing oral secretions and serum IL-17 level and inferior in increasing CD4^(+)/CD8^(+)ratio to DON.It is suggested that both polysaccharides

关 键 词：铁皮石斛 粗多糖部位 非多糖部位 慢性咽炎 T淋巴细胞 NF-κB P65 

分 类 号：R285.5[医药卫生—中药学]