Biopharmaceutical Lab, Department of Biotechnology, School of Bioengineering, SRM Institute of Science and Technology, Kattankulathur, Chennai 603 203, Tamil Nadu, India.
Bioinformatics & Entomoinformatics Lab, School of Bioengineering, SRM Institute of Science and Technology, Kattankulathur, Chennai 603 203, Tamil Nadu, India.
Int Immunopharmacol. 2021 Jan;90:107140. doi: 10.1016/j.intimp.2020.107140. Epub 2020 Nov 11.
The Meretrix meretrix is abundantly present in the Indian coastal areas which can be used as an important useful bioactive source for industrial applications. The M. meretrix visceral mass (MMV) was hydrolysed with four different enzymes and verified for anti-inflammatory activity with the help of HRBC membrane stabilization (HMS) and albumin denaturation (AD) assay. Among the hydrolysates, the tryptic 6th hour hydrolysate was selected for purification using ultrafiltration and size-exclusion chromatography (SEC). Further, the purified peptide was identified to have six amino acid sequence (HKGQCC, 675.582 Da). However, to confirm the anti-inflammatory effects of the purified peptide, it was investigated for nitric oxide synthase (iNOS), pro-inflammatory cytokines production as well as cyclooxygenase-2 (COX-2) activation in lipopolysaccharide (LPS)-stimulated RAW264.7 cells and also evaluated for its functional properties. The in-vitro gastrointestinal digestion was performed on the peptide which cleaved the peptide into two i.e. MMV1 (HK, 284.1 Da) and MMV2 (GQCC, 410.1 Da). The data suggested that the MMV2 peptide have maximum activity and was found to be stable at high temperatures. The MMV2 peptide demonstrated abrupt localization throughout the adult zebrafish body and successfully downregulated the mRNA levels of inflammation-related genes in LPS-induced adult zebrafish. This study indicates that the peptide MMV2 possesses anti-inflammatory activity by suppressing the induced inflammation and can be a strong competitor against non-steroidal anti-inflammatory drugs (NSAIDs).
贻贝（Meretrix meretrix）在印度沿海地区大量存在，可作为工业应用的重要生物活性来源。采用 HRBC 膜稳定（HMS）和白蛋白变性（AD）试验，用四种不同的酶对贻贝内脏质量（MMV）进行水解，并验证其抗炎活性。在水解产物中，选择胰蛋白酶 6 小时水解产物进行超滤和尺寸排阻色谱（SEC）纯化。进一步对纯化肽进行鉴定，发现其具有 6 个氨基酸序列（HKGQCC，675.582 Da）。然而，为了确认纯化肽的抗炎作用，研究了其对脂多糖（LPS）刺激的 RAW264.7 细胞中一氧化氮合酶（iNOS）、促炎细胞因子产生以及环氧化酶-2（COX-2）激活的影响，并评价了其功能特性。对肽进行了体外胃肠消化，将肽裂解成两个肽，即 MMV1（HK，284.1 Da）和 MMV2（GQCC，410.1 Da）。数据表明，MMV2 肽具有最大的活性，并且在高温下稳定。MMV2 肽在成年斑马鱼体内呈现出明显的定位，并且成功下调了 LPS 诱导的成年斑马鱼中炎症相关基因的 mRNA 水平。这项研究表明，肽 MMV2 通过抑制诱导的炎症具有抗炎活性，并且可以成为非甾体抗炎药（NSAIDs）的有力竞争者。
