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为明确酪酪肽(peptide YY,PYY~(3-36))对脂多糖(lipopolysaccharide,LPS)诱导的BV-2细胞中促炎细胞因子及促炎蛋白酶类的影响,本试验用不同浓度的PYY~(3-36)预处理后,再用LPS刺激BV-2细胞,利用MTT法检测PYY~(3-36)的细胞毒作用;分别用荧光定量RT-PCR和Western blot方法检测促炎细胞因子(IL-1β、IL-6和TNF-α)及促炎蛋白酶类(iNOS和COX-2)基因水平和蛋白水平表达变化;分别利用NO检测试剂盒和ELISA试剂盒检测iNOS和COX-2的主要产物NO和PGE2;利用PCR方法检测PYY~(3-36)受体表达情况。结果显示,PYY~(3-36)(10-9~10-11 mol/L)呈浓度依赖性抑制LPS在BV-2细胞中诱导的促炎细胞因子和促炎蛋白酶类的基因和蛋白水平,并能浓度依赖性抑制NO和PGE2。PCR结果表明BV-2细胞中PYY~(3-36)受体Y1、Y2、Y5和Y6表达明显,并且LPS刺激后Y2R受体表达增加。表明PYY~(3-36)在BV-2细胞中能抑制LPS诱导的促炎细胞因子和促炎蛋白酶类的表达以及促炎酶类产物的生成,这个过程可能涉及到其受体Y2。
In order to clarify the effect of peptide YY (PYY 3-36) on proinflammatory cytokines and pro-inflammatory proteases in BV-2 cells induced by lipopolysaccharide (LPS) After pretreatment with PYY ~ (3-36), the BV-2 cells were stimulated with LPS and the cytotoxicity of PYY ~ (3-36) was detected by MTT assay. The proinflammatory cytokines were detected by RT-PCR and Western blot respectively The expression of IL-1β, IL-6 and TNF-α, and the expression of iNOS and COX-2 mRNA and protein were detected by enzyme linked immunosorbent assay. NO and COX- 2, the main products of NO and PGE2; PYY ~ (3-36) receptor expression detected by PCR method. The results showed that PYY ~ (3-36) (10-9 ~ 10-11 mol / L) inhibited the gene and protein levels of proinflammatory cytokines and proinflammatory proteases induced by LPS in BV-2 cells in a concentration-dependent manner , And can inhibit NO and PGE2 in a concentration-dependent manner. The results of PCR showed that the expression of Y1, Y2, Y5 and Y6 of PYY ~ (3-36) receptors in BV-2 cells was significant, and the expression of Y2R receptor increased after LPS stimulation. The results showed that PYY ~ (3-36) could inhibit LPS-induced expression of proinflammatory cytokines and proinflammatory proteases and pro-inflammatory enzymes in BV-2 cells, which may involve its receptor Y2.