【作者】 张程程；

【导师】 徐永平；

【作者基本信息】 大连理工大学 ， 生物工程， 2011， 硕士

【摘要】 鹿角盘有很高的药用价值,民间常用其来治疗乳腺炎、儿童腮腺炎和疮疖等。临床应用也表明鹿角盘具有良好的抗炎作用。但是目前还没有关于鹿角盘抗炎机理的研究报道,因此本论文拟通过炎症动物模型和细胞水平的研究来探讨鹿角盘提取物的抗炎功效及机理。1.采用二甲苯致小鼠耳肿胀模型评价相同生药量的鹿角盘微粉悬液(2.57 g/kg/d)、鹿角盘醇提物(0.12 g/kg/d)和鹿角盘水提物(0.44 g/kg/d)的抗炎效果,以氢化可的松(6mg/kg/d)为阳性对照(用药量皆参考《中国药典》)。正常小鼠灌胃给药十天后,给小鼠右耳涂抹二甲苯建小鼠耳肿胀模型。结果表明：微粉悬液组的肿胀率为132.46±15.26%,与空白对照组的肿胀率170.19±24.68%相比显著降低(P<0.05)；鹿角盘水提物组和氢化可的松组的肿胀率分别为65.43±11.95%和64.63±11.73%,与空白对照组相比极显著降低(P<0.01)；醇提物组肿胀率为154.03±22.43%,与空白对照组相比无显著差异。2.采用角叉菜胶致小鼠足跖肿胀模型来研究鹿角盘水提物的抗炎功效。正常小鼠灌胃给药十天后,给小鼠右后足注射角叉菜胶建小鼠足跖肿胀模型。结果表明：水提物高剂量组(0.44 g/kg/d)和氢化可的松组(6 mg/kg/d)小鼠的足跖肿胀率分别为43.68±2.32%、40.50±2.45%,与空白对照组(等体积生理盐水)小鼠的足跖肿胀率52.91±3.66%相比,均有极显著降低(P<0.01)；水提物低剂量组(0.18 g/kg/d)小鼠的足跖肿胀率为48.46±2.22%,与空白对照组相比显著降低(P<0.05)。表明鹿角盘水提物对角叉菜胶引起的小鼠足跖肿胀有很好的消炎作用。检测小鼠血清和炎足浸出液中的炎症因子,表明鹿角盘水提物可以降低血清中NO、MDA的含量；降低炎足浸出液中PGE2含量；并增强血清中LZM(溶菌酶)、SOD(超氧化物歧化酶)和GSH-PX(谷胱甘肽过氧化物酶)的活力。因此,鹿角盘水提物的抗炎作用可能是：通过抑制NO的过量表达来减少细胞损伤；增强LZM的活力来改善小鼠体内的黏多糖代谢；增强SOD和GSH-PX的活力来减少脂质过氧化,从而减轻炎症反应。另外,鹿角盘水提物能够显著降低足跖肿胀小鼠炎足浸出液中IL-6的含量,而不影响IFN-y和IL-2的含量。说明鹿角盘水提物可能是通过抑制巨噬细胞相关炎症介质的表达来发挥抗炎作用的。3.将LPS诱导的RAW264.7细胞在鹿角盘水提物浓度为0-40 mg/mL的细胞培养液中培养12h,检测细胞培养液中NO、PGE2、IL-6的含量结果表明,随着鹿角盘水提物浓度的增加,细胞培养液中NO、PGE2、IL-6的含量逐渐降低,呈剂量依赖关系。研究表明,鹿角盘水提物有良好的抗炎作用,其抗炎作用可能是通过抑制机体炎症介质过量表达,抑制巨噬细胞相关细胞因子的表达和增强机体内相关酶的活力来实现的。更多还原

【Abstract】 Antler Base has a high medicinal value. Over the past years people used it to treat mastitis, mumps and boils. Clinical applications have confirmed that Antler Base has sound anti-inflammatory effect. But there is no report for its anti-inflammatory mechanism. So this paper proposed to study the anti-inflammatory effect and its mechanism of Antler Base Extract by animal models and cell experiments.1. Xylene-induced ear edema mice model was used to evaluate the anti-inflammatory effect of Antler Base powder suspension(2.57 g/kg/d), ethanol extract of Antler Base(0.12 g/kg/d) and aqueous extract of Antler Base(0.44 g/kg/d), hydrocortisone(6 mg/kg/d) as a positive control. The results showed that Antler Base powder suspension group had lower swelling rate(132.46±15.26%) than the control(170.19±24.68%)(P<0.05). Aqueous extract group and hydrocortisone group had lower swelling rate (65.43±11.95%&64.63±11.73%) than the control (P<0.01). But ethanol extract didn’t have this effect.2. Carrageenan-induced paw edema mice model is one of the most classical models of anti-inflammatory drug research. This model was used to study the anti-inflammatory effect of aqueous extract of Antler Base. The results showed that high-doses aqueous extract of Antler Base group (0.44 g/kg/d) and hydrocortisone group (6 mg/kg/d) had lower swelling rate (43.68±2.32%&40.50±2.45%) than the control 52.91±3.66%(P＜0.01). Low-doses aqueous extract of Antler Base group (0.18 g/kg/d) had lower swelling rate (48.46±2.22%) than the control (P＜0.05).Aqueous extract of Antler Base could also reduce the content of serum NO and MDA, reduce the content of PGE2 in leachate of inflammation foot and enhanc the activity of LZM, SOD and GSH-PX in serum. Its anti-inflammatory effects is probably by inhibiting the over-expression of NO to reduce cell trauma, enhancing LZM activity to improve the metabolism of polysaccharides, enhancing SOD and GSH-PX activity to reduce lipid peroxidation to reduce tissue damage and promote inflammatory tissue injury and recuperate. Aqueous extract of Antler Base could significantly reduce the content of IL-6 in leachate of inflammation foot, but it didn’t affect IFN-y and IL-2 levels significantly. The results showed that aqueous extract of Antler Base might be playing anti-inflammatory effects by inhibiting monocyte-macrophage activation.3. Cultured·the LPS-induced RAW264.7 cells at 0-40 mg/mL concentration of aqueous extract of Antler Base for 12h, then analysis the content of NO,PGE2 and IL-6 in cell culture medium. The results showed that as the concentration of aqueous extract of Antler Base increase, the content of NO, PGE2 and IL-6 in cell culture medium decreased in a dose-dependent manner.The results show that aqueous extract of Antler Base has significant anti-inflammation effect. This effect may occur by inhabiting the body’s over-expression of inflammatory mediators, inhabiting of macrophage-related cytokines and enhancing the activity of the related enzymes of body.更多还原