论文部分内容阅读
目的探讨乳杆菌细胞壁成分(Lactobacilluscell wall extract,LCWE)对小鼠阴道组织β-防御素2诱导表达的影响。方法将雌性ICR小鼠随机分成5组,实验组给予不同浓度的(5、20和100 mg/L)LCWE 200μL阴道接种处理;阴性对照组小鼠阴道接种生理盐水,阳性对照组小鼠阴道接种大肠埃希菌EPEC104菌液(3×109CFU/mL),5 d后处死,取小鼠阴道组织,做阴道菌群分析评价,部分阴道组织提取总RNA,以β-actin为内参照,采用实时荧光定量PCR和Western blot方法检测小鼠阴道组织β-防御素2表达。结果 20 mg/L和100 mg/L浓度LCWE处理后,小鼠阴道菌群较生理盐水组的肠杆菌、葡萄球菌明显减少(P<0.05),乳杆菌无显著性变化(P>0.05)。生理盐水组小鼠阴道组织β-防御素2 mRNA表达量极低,大肠埃希菌阳性对照组能显著诱导小鼠阴道组织β-防御素2 mRNA的表达;5 mg/L浓度LCWE处理组开始,β-防御素2表达量开始提高,在100 mg/L浓度LCWE组其表达量达到最高。Western blot结果显示:生理盐水组小鼠阴道组织未检测到β-防御素2蛋白表达,而100 mg/L浓度LCWE处理组和大肠埃希菌EPEC104组均出现特异性条带。结论 LCWE可上调小鼠阴道组织β-防御素2的表达且存在着剂量依赖关系。
Objective To investigate the effect of Lactobacilluscell wall extract (LCWE) on the induced expression of β-defensin 2 in vaginal tissues of mice. Methods Female ICR mice were randomly divided into five groups. The experimental group was given vaginal inoculation with different concentrations (5, 20 and 100 mg / L) of LCWE 200μL. Negative control mice were inoculated vaginally with saline and the positive control mice were inoculated vaginally Escherichia coli EPEC104 bacteria solution (3 × 109CFU / mL), 5 d after sacrifice, take the vagina tissue of mice to do vaginal flora analysis and evaluation, part of the vaginal tissue total RNA extracted with β-actin as internal reference, real-time Fluorescence quantitative PCR and Western blot were used to detect the expression of β-defensin 2 in vaginal tissues of mice. Results After treated with LCWE of 20 mg / L and 100 mg / L, the vaginal flora of mice was significantly decreased (P <0.05) and that of Lactobacillus was no significant difference (P> 0.05). The expression of β-defensin 2 mRNA in the vaginal tissue of the normal saline group was extremely low, and the positive control group of Escherichia coli induced the expression of β-defensin 2 mRNA in the vaginal tissue of the mice. LCWE 5 mg / L , the expression of β-defensin 2 began to increase, and reached the highest level in 100 mg / L LCWE group. Western blot results showed that the expression ofβ-defensin 2 was not detected in the vaginal tissues of the normal saline group, while the specific bands appeared in both the LCWE 100 mg / L and the EPEC104 Escherichia coli groups. Conclusion LCWE can up-regulate the expression of β-defensin 2 in vaginal tissues in a dose-dependent manner.