豌豆ACE抑制肽对诱导损伤的EA.hy.926细胞的保护作用

Protective effects of the pea ACE inhibitory peptide on induced damage of EA.hy.926 cells

目的:实现豌豆蛋白的高值化利用。方法:以豌豆蛋白为原料,采用双酶协同酶解法制备豌豆蛋白源ACE抑制肽,以酶解时间和酶添加顺序为变量,以ACE抑制率、水解度、可溶蛋白含量为指标确定豌豆ACE抑制肽的最佳酶解工艺。将最佳酶解工艺条件下制得的ACE抑制肽进行超滤分级,探究分子量对ACE抑制活性的影响,最后通过测定ET-1、MDA含量、SOD和NO含量水平验证ACE抑制活性较高的豌豆肽对诱导损伤的EA.hy.926细胞的保护作用。结果:双酶法制备豌豆ACE抑制肽的最佳工艺条件为底物质量浓度100 mg/mL,3.0%的碱性蛋白酶在pH 9.5、50℃条件下酶解2.0 h后加入3.0%的复合蛋白酶,在pH 7.0、50℃条件下继续酶解2.0 h。此条件下的豌豆蛋白水解物ACE抑制率的IC_(50)值为1.141 mg/mL;小于2.5 kDa的肽组分其ACE抑制活性最强,且能显著减少Ang-Ⅱ诱导损伤细胞的ET-1、MDA含量,增加SOD和NO含量水平。结论:豌豆ACE抑制肽对Ang-Ⅱ诱导损伤EA.hy.926细胞具有保护作用。

Objective:This study aimed to realize the high-value utilization of pea protein.Methods:Using pea protein as raw material,ACE inhibitory peptide derived from pea protein was prepared by double-enzyme coordinated enzymatic hydrolysis method,and the ACE inhibition rate,hydrolysis degree and soluble protein content of the hydrolysate were used as indicators to determine the best enzymatic process for the pea ACE inhibitory peptide.Then the ACE inhibitory peptides produced under the optimal enzymatic digestion process were graded by ultrafiltration to explore the effect of molecular weight on the ACE inhibitory activity.Finally,the protective effect of the pea peptide with high ACE inhibitory activity against the injured EA.hy.926 cells was verified by measuring ET-1,MDA content,SOD and NO content levels.Results:The optimal preparation conditions of pea ACE inhibitory peptide by double-enzyme method were as follows:substrate concentration of 100 mg/mL,3.0%alcalase enzymolysis for 2.0 h at pH 9.5 and 50℃,and then adding 3.0%compound protease for 2.0 h at pH 7.0 and 50℃.Under these conditions,the IC_(50) value of ACE inhibition rate of pea protein hydrolysate was 1.141 mg/mL.Peptide components less than 2.5 kDa had the strongest ACE inhibitory activity and could significantly reduce ET-1,MDA content and increase SOD and NO content levels of Ang-induced injured cells.Conclusion:The pea ACE inhibitory peptide was protective against Ang-induced damage to EA.hy.926 cells.