金银花对金黄色葡萄球菌致山羊乳腺上皮细胞损伤的修复作用
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摘要
金银花在家畜乳房炎中有广泛的应用，但是关于其如何防治乳房炎、对乳腺上皮细胞作用的研究少见，为研究其相关及预防、治疗机制，本试验分别观察金银花水提取物、金黄色葡萄球菌、金银花水提取物及金黄色葡萄球菌共同对体外培养的山羊乳腺上皮细胞的作用，为乳房炎的预防治疗提供相关资料。
本试验主要采用分子生物学方法检测不同处理后山羊乳腺上皮细胞分泌标志性蛋白，β-casein的蛋白及基因水平的表达变化。试剂盒检测乳腺上皮细胞损伤标志性酶活力的变化。包括以下三个方面：①不同浓度中药金银花对山羊乳腺上皮细胞体外增殖和β-casein表达的影响，②不同量的金黄色葡萄球菌裂解液对体外山羊乳腺上皮细胞不同时间的细胞活性、细胞凋亡、western-bloting及实时荧光定量检测β-casein的蛋白和mRNA表达量、NAG-ase、LDH、MPO酶活力的变化。③金银花和不同量的金黄色葡萄球菌裂解液共同对山羊乳腺上皮细胞的细胞凋亡、酶活力及β-casein的表达变化。获得以下实验结果：
1金银花水提取物在1000μg/ml时能极显著提高细胞活性和β-casein的表达（P<0.01）。
2不同量的金黄色葡萄球菌裂解液对乳腺上皮细胞有损伤作用，同一时间裂解液量越高、同一剂量的时间越长，细胞呈现活性下降，凋亡率升高、β-casein表达下降，NAG-ase、LDH、MPO酶活力升高的变化（P<0.01）。
31000μg/ml金银花水提取物能抑制金黄色葡萄球菌裂解液所引起的乳腺上皮细胞β-casein的表达降低、凋亡率升高和酶活力升高的变化。
以上试验结果说明金黄色葡萄球菌可以引起乳腺上皮细胞损伤的炎性变化，金银花提取物能够有效的修复损伤的炎性变化。
The mastitis is a very harmful disease on milk production,which is highlyincidence and has a low cure rate. In order to study the mechanism to prevention andtreatment of mastitis, we observed interaction of the extracts of Lonicera japonica andStaphylococcus aureus on mammary epithelial cells to provide information. So we tookmethods of molecular biology to research the effects on goat mammary epithelial cellsof Lonicera japonica and Staphylococcus aureus.This experiment mainly includes thefollowing three aspects:(1) different concentrations of traditional Lonicera japonicaeffect the milk protein on goats mammary epithelial cells in vitro (2) goat mammaryepithelial cells have toxicity action to different amount of staphylococcus aureus liysisbuffer in vitro.(3) Lonicera japonica suppression inflammatory reaction which initiateby different amount of staphylococcus aureus liysis buffer on goat mammary epithelialcells in vitro. We got the following results:
1Lonicera japonica extract at the concentration of1000μg/ml significantlypromoted the expression of beta casein (P <0.01).
2Different amount of staphylococcus aureus liysis buffer influence expression ofgoat mammary epithelial cells,the expression of beta-casein was significant highcompared with control group when trained after48and72hours,the same change withthe three symbol enzymes of goat mammary epithelial cells damaged (P <0.01).
3Different amount of staphylococcus aureus liysis buffer together with Lonicerajaponica extract at the concentration of1000μg/ml influence three symbol enzymes ofgoat mammary epithelial cells damaged, which was obviously lower than no Lonicerajaponica joined. It said that the Lonicera japonica has the effect to cure the damage ongoat mammary epithelial cells.
The experiment results show that the staphylococcus aureus can causeinflammatory of goat mammary epithelial cells, Lonicera japonica extract caneffectively restrain the inflammatory caused by staphylococcus aureus.
本试验主要采用分子生物学方法检测不同处理后山羊乳腺上皮细胞分泌标志性蛋白，β-casein的蛋白及基因水平的表达变化。试剂盒检测乳腺上皮细胞损伤标志性酶活力的变化。包括以下三个方面：①不同浓度中药金银花对山羊乳腺上皮细胞体外增殖和β-casein表达的影响，②不同量的金黄色葡萄球菌裂解液对体外山羊乳腺上皮细胞不同时间的细胞活性、细胞凋亡、western-bloting及实时荧光定量检测β-casein的蛋白和mRNA表达量、NAG-ase、LDH、MPO酶活力的变化。③金银花和不同量的金黄色葡萄球菌裂解液共同对山羊乳腺上皮细胞的细胞凋亡、酶活力及β-casein的表达变化。获得以下实验结果：
1金银花水提取物在1000μg/ml时能极显著提高细胞活性和β-casein的表达（P<0.01）。
2不同量的金黄色葡萄球菌裂解液对乳腺上皮细胞有损伤作用，同一时间裂解液量越高、同一剂量的时间越长，细胞呈现活性下降，凋亡率升高、β-casein表达下降，NAG-ase、LDH、MPO酶活力升高的变化（P<0.01）。
31000μg/ml金银花水提取物能抑制金黄色葡萄球菌裂解液所引起的乳腺上皮细胞β-casein的表达降低、凋亡率升高和酶活力升高的变化。
以上试验结果说明金黄色葡萄球菌可以引起乳腺上皮细胞损伤的炎性变化，金银花提取物能够有效的修复损伤的炎性变化。
The mastitis is a very harmful disease on milk production,which is highlyincidence and has a low cure rate. In order to study the mechanism to prevention andtreatment of mastitis, we observed interaction of the extracts of Lonicera japonica andStaphylococcus aureus on mammary epithelial cells to provide information. So we tookmethods of molecular biology to research the effects on goat mammary epithelial cellsof Lonicera japonica and Staphylococcus aureus.This experiment mainly includes thefollowing three aspects:(1) different concentrations of traditional Lonicera japonicaeffect the milk protein on goats mammary epithelial cells in vitro (2) goat mammaryepithelial cells have toxicity action to different amount of staphylococcus aureus liysisbuffer in vitro.(3) Lonicera japonica suppression inflammatory reaction which initiateby different amount of staphylococcus aureus liysis buffer on goat mammary epithelialcells in vitro. We got the following results:
1Lonicera japonica extract at the concentration of1000μg/ml significantlypromoted the expression of beta casein (P <0.01).
2Different amount of staphylococcus aureus liysis buffer influence expression ofgoat mammary epithelial cells,the expression of beta-casein was significant highcompared with control group when trained after48and72hours,the same change withthe three symbol enzymes of goat mammary epithelial cells damaged (P <0.01).
3Different amount of staphylococcus aureus liysis buffer together with Lonicerajaponica extract at the concentration of1000μg/ml influence three symbol enzymes ofgoat mammary epithelial cells damaged, which was obviously lower than no Lonicerajaponica joined. It said that the Lonicera japonica has the effect to cure the damage ongoat mammary epithelial cells.
The experiment results show that the staphylococcus aureus can causeinflammatory of goat mammary epithelial cells, Lonicera japonica extract caneffectively restrain the inflammatory caused by staphylococcus aureus.
引文
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