中草药提取物对东北林蛙源嗜水气单胞菌体内外抑菌试验及其抗炎免疫活性研究  

Antibacterial activity anti-inflammatory and immune activity of Chinese herbal extracts against Aeromonas hydrophila from northeastern forest frogs in vivo and in vitro

作　　者：孟欣 孙大庆 张冬星[1] 单晓枫[1] 钱爱东[1] MENG Xin;SUN Daqing;ZHANG Dongxing;SHAN Xiaofeng;QIAN Aidong(College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China)

机构地区：[1]吉林农业大学动物科学技术学院,长春130118

出　　处：《黑龙江畜牧兽医》2022年第14期112-118,123,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：吉林省教育厅项目(JJKH20210367KJ)。

摘　　要：为了研究五倍子、红花、黄连、地榆、射干、黄芪六味中药对嗜水气单胞菌的体内外抑菌作用,试验采用牛津杯打孔法和二倍稀释法测定嗜水气单胞菌对六味中药提取物的敏感性,以及中药提取物的最小抑菌浓度(MIC)和最小杀菌浓度(MBC),筛选出能够有效抑制嗜水气单胞菌的中药提取物,并选取健康的东北林蛙为试验对象进行体内试验,选取抑菌效果较好的中药提取物混合饲料对试验林蛙进行饲喂,共饲喂56 d,于饲喂第0,14,28,42,56天取样,检测在中药混合饲料饲喂后林蛙机体相关免疫指标[血清中酸性磷酸酶(AKP)、超氧化物歧化酶(SOD)和溶菌酶(LZM)的活性,心脏、脾脏、肺脏组织中白细胞介素-10(IL-10)、白细胞介素1β(IL-1β)、肿瘤坏死因子(TNF-α)和γ干扰素(IFN-γ)基因mRNA相对表达量]变化及嗜水气单胞菌攻毒后林蛙的存活率。结果表明:六味中药中,供试菌对五倍子、红花、黄连表现为中度敏感,五倍子、红花、黄连、地榆、射干、黄芪对嗜水气单胞菌的MIC分别是0.05,0.10,0.10,0.20,0.25,0.20 g/mL,MBC分别是0.10,0.15,0.25,0.25,0.40,0.40 g/mL。五倍子、红花对试验菌株的抑菌作用较强。综合体外抑菌试验结果,选择五倍子和红花两味中药进行体内试验,分为对照组、五倍子组和红花组。与对照组相比,五倍子组和红花组饲喂2周后均可使林蛙血清中AKP、SOD和LZM活性上升;心脏、肝脏和脾脏(除红花组脾脏组织第42天外)中IL-10、IL-1β、TNF-α和IFN-γ基因表达水平均有不同程度升高。五倍子和红花可有效缓解嗜水气单胞菌对东北林蛙的侵袭。说明五倍子和红花两味中药可对嗜水气单胞菌产生抑制作用,并且对感染嗜水气单胞菌的东北林蛙产生免疫调节作用。In order to study the in vitro and in vivo antibacterial effects of six Chinese herbal medicines,namely,Chinese gallnut,safflower,golden thread,garden burnet root,blackberry lily rhizome and milkvetch root on Aeromonas hydrophila,the sensitivity of Aeromonas hydrophila to the six herbal extracts was measured by Oxford cup drilling method and two-fold dilution method.The minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of the herbal extracts were selected to effectively inhibit the bacteria.Healthy northeastern forest frogs were selected as the test subjects for the in vivo test,and the frogs were fed with the herbal mixture with good antibacterial effect for 56 d.Samples were taken on days 0,14,28,42 and 56,and the immune indexes(acid phosphatase[AKP],superoxide dismutase[SOD]and lysozyme[LZM]activities in serum,and the relative mRNA expression of interleukin-10[IL-10],interleukin-1β[IL-1β],tumor necrosis factor-α[TNF-α]and interferon-γ[IFN-γ]genes in heart,spleen and lung tissues)were detected and the survival rate of Aeromonas hydrophila was determined.The results showed that among the six traditional Chinese medicines,the tested bacteria were moderately sensitive to Chinese gallnut,safflower and golden thread.The MIC of Chinese gallnut,safflower,golden thread,garden burnet root,blackberry lily rhizome and milkvetch root were 0.05,0.10,0.10,0.20,0.25,0.20 g/mL,and MBC were 0.10,0.15,0.25,0.25,0.40,0.40 g/mL,respectively.Chinese gallnut and safflower had strong antibacterial effect on the tested strains.Based on the results of bacteriostatic test in vitro,Chinese gallnut and safflower were divided into control group,Chinese gallnut group and safflower group.Compared with the control group,the activities of AKP,SOD and LZM in serum in gallnut group and safflower group were increased after feeding for 2 weeks.The expression levels of IL-10,IL-1β,TNF-αand IFN-γgenes in the heart,liver and spleen(except for the tissue of spleen in safflower group on days 42)were increased to d

关 键 词：东北林蛙 嗜水气单胞菌 中草药 五倍子 红花 免疫调节 

分 类 号：S853.7[农业科学—临床兽医学]