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

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 m L的1×108CFU/mL LP-YS2、1×10~9CFU/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n NOS、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。

In this study, the mouse colitis model was induced by dextran sulfate sodium （ DSS）, and the inhibitory effects of LactobaciUus plautarum Y S-2 （LP-YS2 ）were observed.The experimental mice were divided into non-aal 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 x 10s CFU/mL LP-YS2,1 × 10^9 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 ）, ehemokine （ 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 signifieantly（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, e-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 eoul be seen that LP-Y$2 had the effects of colitis suppression.