Department of Molecular Biology, Ahar Branch, Islamic Azad University, Ahar, Iran.
J Pept Sci. 2014 Jun;20(6):429-37. doi: 10.1002/psc.2634. Epub 2014 Apr 10.
Human catestatin CgA352-372 (SL21) is an endogenous neuropeptide with multiple biological functions. The present study aimed to evaluate the antioxidant, antibacterial, cytotoxic, and DNA damage protective effects of SL21 neuropeptide. SL21 neuropeptide generated from the C-terminus of chromogranin A (CgA) was synthesized by solid-phase method. Synthetic peptide was subjected to various in vitro antioxidant assays including the scavenging of 1,1-diphenyl-2-pycryl-hydrazyl (DPPH), 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS(·+) ), and hydroxyl free radicals, metal ion chelation, inhibition of lipid peroxidation, and reducing power. Moreover, protective effect of SL21 on H2 O2 -induced DNA damage was analyzed using pTZ57/RT plasmid. Methylthiazoltetrazolium assay was also performed to study the cytotoxic effect of SL21 neuropeptide on human peripheral blood mononuclear cells. Furthermore, antibacterial and hemolysis assays were conducted. The results demonstrated high activities of SL21 in scavenging free radicals (DPPH, ABTS(·+) , and hydroxyl), chelating of Cu(2+) /Fe(2+) metal ions, reducing power, and inhibition of lipid peroxidation in a concentration-dependent manner. SL21 neuropeptide revealed a protective effect on DNA damage caused by hydroxyl radicals. Interestingly, the peptide exhibited no significant cytotoxicity towards peripheral blood mononuclear cells. Furthermore, SL21 peptide displayed antimicrobial activity against Staphylococcus aureus and Pseudomonas aeruginosa without any hemolytic activity on human red blood cells. Conclusively, the present study established SL21 (catestatin) as a novel antioxidative peptide that could further be investigated for its potential use as a pharmaceutical agent.
人嗜铬粒蛋白 CgA352-372(SL21)是一种具有多种生物学功能的内源性神经肽。本研究旨在评估 SL21 神经肽的抗氧化、抗菌、细胞毒性和 DNA 损伤保护作用。SL21 神经肽是通过固相法从嗜铬粒蛋白 A(CgA)的 C 端合成的。合成的肽进行了各种体外抗氧化测定,包括 1,1-二苯基-2-苦基肼基(DPPH)、2,2-联氮-双(3-乙基苯并噻唑啉-6-磺酸)(ABTS(·+))和羟基自由基的清除、金属离子螯合、抑制脂质过氧化和还原能力。此外,还使用 pTZ57/RT 质粒分析了 SL21 对 H2O2 诱导的 DNA 损伤的保护作用。噻唑蓝(MTT)测定法也用于研究 SL21 神经肽对人外周血单个核细胞的细胞毒性作用。此外,还进行了抗菌和溶血测定。结果表明,SL21 在清除自由基(DPPH、ABTS(·+)和羟基)、螯合 Cu(2+)/Fe(2+)金属离子、还原能力和抑制脂质过氧化方面具有很高的活性,且呈浓度依赖性。SL21 神经肽对羟自由基引起的 DNA 损伤具有保护作用。有趣的是,该肽对人外周血单个核细胞没有明显的细胞毒性。此外,SL21 肽对金黄色葡萄球菌和铜绿假单胞菌具有抗菌活性,且对人红细胞无溶血活性。综上所述,本研究确立了 SL21(嗜铬粒蛋白)作为一种新型抗氧化肽,可进一步研究其作为药物制剂的潜力。