Hidayat Meilinah, Prahastuti Sijani, Yusuf Muhammad, Hasan Khomaini
Faculty of Medicine, Universitas Kristen Maranatha, Jalan Prof. Drg. Suria Sumantri 65 Bandung 40163, Indonesia.
Research Center for Molecular Biotechnology and Bioinformatics, Universitas Padjadjaran, Jalan Singaperbangsa No. 2, Bandung 40133, Indonesia.
Iran J Basic Med Sci. 2021 Jun;24(6):734-743. doi: 10.22038/ijbms.2021.50291.11459.
Fibrosis is the major cause of chronic kidney injury and the primary etiology in diabetic glomerulosclerosis. The initial study of protein hydrolysate of green peas hydrolyzed by bromelain (PHGPB) considered it to improve kidney function parameters and showed no fibrosis in histopathology features in gentamicin-induced nephrotoxicity rats. In the current study, we aimed to assess the nutrition profile and potency of RGD in PHGPB as antifibrosis in chronic kidney disease (CKD).
Green peas () were hydrolyzed by bromelain from pineapple juice to obtain PHGPB. The amino acid content of PHGPB was measured using the UPLC method, while the primary structure used LC-MS/MS. Bioinformatic analysis was conducted using the Protease Specificity Predictive Server (PROSPER). The potency of RGD in PHGPB was characterized by determining the levels of Fibronectin (FN) and TGF-β1 in mesangial SV40 MES 13 cell lines of diabetic glomerulosclerosis.
The level of lysine was 364.85 mg/l. The LC-MS/MS data showed two proteins with 4-15 kDa molecular weight originated from convicilin (P13915 and P13919) which were predicted by PROSPER proteolytic cleavage, resulted in RGD in the LERGDT sequence peptide. PHGPB increased SV40 MES 13 mesangial cell proliferation that died from high-glucose levels (diabetic glomerulosclerosis model). PHGPB and RGD reduced the levels of FN and TGF-β1 in mesangial cell lines of diabetic glomerulosclerosis.
The nutrition profile and RGD motif in PHGPB show great potential as antifibrosis in CKD.
纤维化是慢性肾损伤的主要原因，也是糖尿病肾小球硬化症的主要病因。对菠萝蛋白酶水解绿豌豆所得蛋白水解物（PHGPB）的初步研究认为其可改善肾功能参数，且在庆大霉素诱导的肾毒性大鼠的组织病理学特征中未显示纤维化。在本研究中，我们旨在评估PHGPB中RGD的营养成分和作为慢性肾脏病（CKD）抗纤维化剂的效力。
用菠萝汁中的菠萝蛋白酶水解绿豌豆以获得PHGPB。采用超高效液相色谱法测定PHGPB的氨基酸含量，而一级结构则采用液相色谱-串联质谱法。使用蛋白酶特异性预测服务器（PROSPER）进行生物信息学分析。通过测定糖尿病肾小球硬化症系膜SV40 MES 13细胞系中纤连蛋白（FN）和转化生长因子-β1（TGF-β1）的水平来表征PHGPB中RGD的效力。
赖氨酸水平为364.85毫克/升。液相色谱-串联质谱数据显示有两种分子量为4-15 kDa的蛋白质源自伴刀豆球蛋白（P13915和P13919），经PROSPER蛋白水解切割预测，在LERGDT序列肽中产生了RGD。PHGPB增加了因高糖水平死亡的SV40 MES 13系膜细胞增殖（糖尿病肾小球硬化症模型）。PHGPB和RGD降低了糖尿病肾小球硬化症系膜细胞系中FN和TGF-β1的水平。
PHGPB的营养成分和RGD基序在CKD抗纤维化方面显示出巨大潜力。
