鳕鱼蛋白酶解肽不同分子量组分抗炎活性的比较研究
Comparison of anti- inflammatory activity of four different molecular fractions of cod protein hydrolytic peptides
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摘要: 本研究比较了鳕鱼蛋白酶解肽（Cop protein hydrolytic peptide,CPHP）不同分子量组分对细胞炎症因子释放的抑制作用。鳕鱼蛋白经复合蛋白酶酶解制备得到鳕鱼蛋白酶解肽（CPHP）,并进一步超滤得到三个不同分子量组分CPHP1（>5000 u）、CPHP2（<3000 u）和CPHP3（30005000 u）。用LPS诱导RAW264.7细胞炎症因子释放为模型,分别采用Griess法和ELISA分析NO和TNF-α、IL-1β、IL-6等细胞炎症因子的含量。结果表明,CPHP不同分子量组分对LPS（500μg/m L）所致的RAW264.7细胞生长抑制均有显著保护作用（p<0.05）,其保护作用随CPHP不同分子量组分的浓度升高而逐渐增强;CPHP和CPHP1的保护作用强于CPHP2和CPHP3。CPHP不同分子量组分对LPS（1μg/m L）诱导RAW264.7细胞NO、IL-1β、IL-6和TNF-α等炎症因子释放均有显著的抑制作用（p<0.05）,其抑制作用随CPHP不同分子量组分的浓度升高而逐渐增强;CPHP和CPHP1的抑制作用强于CPHP2和CPHP3。总之,CPHP不同分子量组分对LPS所致的细胞生长抑制均有显著的保护作用（p<0.05）,对LPS诱导的细胞炎症因子的释放均有显著抑制作用（p<0.05）;CPHP和CPHP1的细胞保护作用和炎症因子释放抑制作用均强于CPHP2和CPHP3的。Abstract: Effects of four different molecular fractions of cod protein hydrolytic peptides( CPHP) on LPS- induced release of inflammatory cytokines were compared. CPHP was a whole mixture of cod protein hydrolytic peptides hydrolyzed by complex protease. The peptides were further fractionated by ultrafiltration into three fractions of CPHP1( > 5000 u),CPHP2( < 3000 u) and CPHP3( 3000 ~ 5000 u). Griess assay and ELISA kits were used to measure the levels of NO and inflammatory cytokines( TNF-α,IL-1β and IL-6),respectively. The results showed that the all CPHP fractions protected RAW264.7 cells from LPS( 500 μg / m L) induced cell growth inhibition,in a dose dependent manner,and protective effects of CPHP and CPHP1 were higher than that of CPHP2 and CPHP3. All CPHP fractions inhibited release of NO,IL-1β,IL-6 and TNF-α from RAW264.7 cells induced by LPS( 1 μg / m L),in a dose dependent manner.The inhibitory effects of CPHP and CPHP1 were higher than that of CPHP2 and CPHP3.In conclusion,all four fractions of CPHP had protective effect against LPS induced cell growth inhibition and release of inflammatory cytokines.The CPHP and CPHP1 had higher activity than that of CPHP2 and CPHP3.
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