<i>Lactobacillus plantarum</i> YS-2对葡聚糖硫酸钠诱导C57BL/6J小鼠结肠炎的抑制作用

骞宇; 雷爱玲; 刘晓敬; 易若琨; 赵欣

doi:10.13386/j.issn1002-0306.2018.15.053

Lactobacillus plantarum YS-2对葡聚糖硫酸钠诱导C57BL/6J小鼠结肠炎的抑制作用

Inhibitory Effects of Lactobacillus plantarum YS-2 in Dextran Sulfate Sodium-Induced C57BL/6J Mice Colitis

摘要

摘要: 本研究通过葡聚糖硫酸钠(DSS)诱导小鼠结肠炎模型,观察了Lactobacillus plantarum YS-2(LP-YS2)对结肠炎抑制效果。将实验小鼠分为正常组、模型组、LP-YS2-L组、LP-YS2-H组和柳氮磺胺吡啶组,对除正常组外其余组小鼠诱导结肠炎,同时对LP-YS2-L组、LP-YS2-H组和柳氮磺胺吡啶组小鼠每日分别灌胃0.2 mL的1×10⁸ CFU/mL LP-YS2、1×10⁹ CFU/mL LP-YS2和20 mg/kg柳氮磺胺吡啶,持续5周。然后通过H&E切片观察小鼠结肠形态,通过内皮素(ET)、生长抑素(SS)、P物质(SP)、血管活性肠肽(VIP)、髓过氧化物酶(MPO)、谷胱甘肽(GSH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、白介素-2(IL-2)、白介素-10(IL-10)试剂盒检测小鼠的血清和结肠组织;同时,采用荧光定量PCR技术对小鼠结肠组织神经元型一氧化氮合酶(nNOS)、内皮型一氧化氮合酶(eNOS)、诱导型一氧化氮合成酶(iNOS)、c-Kit、干细胞因子(SCF)、白介素-8(IL-8)、趋化因子受体2(CXCR2)的mRNA表达进行了检测,同时以药物柳氮磺胺吡啶为阳性对照比较LP-YS2的作用效果。实验结果显示相对于模型组,LP-YS2可以显著(p<0.05)增加结肠炎小鼠结肠长度和提高结肠质量/结肠长度的比值。对小鼠血清进行的检测表明LP-YS2能够显著(p<0.05)降低结肠炎小鼠(模型组)血清ET、SP、IL-10水平和显著(p<0.05)提高SS、VIP、IL-2水平。对小鼠结肠组织的检测显示LP-YS2可以显著(p<0.05)提高结肠炎小鼠(模型组)结肠中GSH含量、SOD活力和降低MPO活力、MDA含量;LP-YS2还能够显著(p<0.05)上调结肠炎小鼠(模型组)结肠中的nNOS、eNOS、c-Kit、SCF mRNA表达和显著(p<0.05)下调iNOS、IL-8、CXCR2表达。由此可以看出,LP-YS2具有结肠炎抑制效果。

Abstract: In this study,the mouse colitis model was induced by dextran sulfate sodium(DSS),and the inhibitory effects of Lactobacillus plantarum YS-2(LP-YS2)were observed. The experimental mice were divided into normal group,model group,LP-YS2-L group,LP-YS2-H group and sulfasalazine group,the other groups except the normal group mice were induced colitis,meanwhile,LP-YS2-L group,LP-YS2-H group and sulfasalazine group mice were administered daily 0.2 mL 1×10⁸ CFU/mL LP-YS2,1×10⁹ CFU/mL LP-YS2 and 20 mg/kg sulfasalazine,for 5 weeks. Then the colonic morphology of mice were determined by H&E section observation,the serum and colon tissues of mice colon were determined using kits endothelin(ET),somatostatin(SS),substance P(SP),vasoactive intestinal peptide(VIP),myeloperoxidase(MPO),glutathione(GSH),malondialdehyde(MDA),superoxide dismutase(SOD),interleukin-2(IL-2),interleukin-10(IL-10),and the mRNA expression of neuronal nitric oxide synthase(nNOS),endothelial nitric oxide synthase(eNOS),inducible nitric oxide synthase(iNOS),c-Kit,stem cell factor(SCF),interleukin-8(IL-8),chemokine(C-X-C motif)receptor 2(CXCR2)were determined by fluorescent quantitative PCR technique. The experiment results showed that compared with the model group,LP-YS2 could significantly(p<0.05)increase colon length and the ratio of colon weight/colon length in colitis mice(model group). The detection of mice serum showed that LP-YS2 could significantly(p<0.05)reduce the serum levels of ET,SP,IL-10,and significantly(p<0.05)increase the levels of SS,VIP and IL-2 in colitis mice(model group). The detection of colon tissue showed that LP-YS2 could significantly(p<0.05)raise the GSH content,SOD activity and significantly(p<0.05)reduce the MPO activity,MDA content in the colon of colitis mice(model group). LP-YS2 could also significantly(p<0.05)up regulate the mRNA expression of nNOS,eNOS,c-Kit,SCF,and significantly(p<0.05)down regulate the expression of iNOS,IL-8,CXCR2 in the colon of colitis mice(model group). From these results,it coul be seen that LP-YS2 had the effects of colitis suppression.