骆驼蓬子活性成分的筛选及小鼠乳腺炎治疗效果评价

Screening of Active Ingredients of Peganum harmala Seed and Evaluation of Therapeutic Effect in Mouse Mastitis

为探明骆驼蓬(Peganum harmala L.)种子的抑菌作用及小鼠乳腺炎的治疗效果。本研究采用溶剂提取和超声辅助提取等方式对骆驼蓬子中的多糖、多酚、生物碱、黄酮类活性成分进行提取;通过琼脂平板扩散法(打孔法)和微量肉汤法对各提取物的抑菌效果进行检测;通过急性毒性预试验和最大耐受量试验对抑菌效果显著的提取物进行安全性检测;最后用于小鼠乳腺炎模型,对其治疗效果进行评价。结果表明,骆驼蓬子中含有多糖、多酚、生物碱、黄酮类活性物质,相应提取物的含量分别为36.90%±0.27%、23.18%±0.36%、22.24%±0.08%、20.46%±0.14%。抑菌试验结果显示,骆驼蓬子多酚和黄酮的抑菌效果较为显著,对大肠杆菌(E.coli)、金黄色葡萄球菌(S.aureus)、无乳链球菌(S.agalactiae)的最小抑菌浓度(MIC)和最小杀菌浓度(MBC)分别在0.7813~1.5625 mg/mL和0.7813~12.5000 mg/mL。安全性试验结果显示,骆驼蓬子多酚和黄酮对小鼠的半数致死量(LD_(50))值均大于1200 mg/kg,其最大耐受量是4.5 g/kg。小鼠乳腺炎治疗结果显示,骆驼蓬子多酚和黄酮能够显著降低乳腺组织中的TNF-α、IL-6含量及菌落数量(P<0.05),减少血管扩张和炎性细胞浸润程度,加快腺泡结构和功能的恢复。其中,骆驼蓬子多酚的作用效果更为显著(P<0.05)。骆驼蓬子多酚不但对引发奶牛乳腺炎的大肠杆菌、金黄色葡萄球菌、无乳链球菌具有较强的抑制效果,而且能够降低金黄色葡萄球所致的乳腺炎小鼠乳腺组织中的菌落数量、IL-6和TNF-α含量,增强乳腺组织修复能力。

To investigate the antibacterial effect of Peganum harmala seed and the therapeutic effect in the mouse mastitis.In this study,the polysaccharides,polyphenols,alkaloids,and flavonoids in Peganum harmala seed were extracted by solvent extraction and ultrasound-assisted extraction;the in vitro antibacterial effects of each extract were examined by plate diffusion method and micro-broth method;the safety of the extracts with significant antibacterial effects was examined by acute toxicity pre-test and maximum tolerance test;Finally,the extracts were used in a mouse mastitis model to evaluate their therapeutic effects.The results showed that Peganum harmala seed contained polysaccharides,polyphenols,alka-loids,and flavonoids,and the contents of the corresponding extracts were 36.90%±0.27%,23.18%±0.36%,22.24%±0.08%and 20.46%±0.14%,respectively.In vitro antibacterial tests showed that Pe-ganum harmalaseed polyphenols and flavonoids had more significant antibacterial effects,and the inhibi-tory effects on E.coli,S.aureus,and S.agalactiae were extremely and highly sensitive,with MIC and MBC ranging from 0.7813-1.5625 mg/mL and 0.7813-12.5000 mg/mL,respectively.The safety test showed that the LD50 values of Peganum harmala seed polyphenols and flavonoids in mice were greater than 1200 mg/kg,and their maximum tolerable dose was 4.5 g/kg.The effect of mastitis treatment in mice showed that Peganum harmala seed polyphenols and flavonoids could significantly reduce the TNF-αand IL-6 contents and the number of colonies in mammary tissue(P<0.05),reduce the degree of vasodi-lation and inflammatory cell infiltration,and accelerate gradual recovery of glandular follicle structure and function.Among them,the effect of Peganum harmala seed polyphenols was more significant(P<0.05).Peganum harmala seed polyphenols not only had strong inhibitory and killing effects on E.coli,S.aureus and S.agalactiae,which caused mastitis in dairy cows,but also it reducd the number of colo-nies,IL-6 and TNF-αcontent in the mammary tissue of mastitis mice caused by S.aureus and enhanced the repair ability of mammary tissue.