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中國精品科技期刊2020
蘭夢,李晶峰,李冬冰,等. 人參不同炮制品蛋白酶解肽對LPS誘導RAW264.7細胞的抗炎作用研究[J]. 食品工業科技,2024,45(9):350?358. doi: 10.13386/j.issn1002-0306.2023050282.
引用本文: 蘭夢,李晶峰,李冬冰,等. 人參不同炮制品蛋白酶解肽對LPS誘導RAW264.7細胞的抗炎作用研究[J]. 食品工業科技,2024,45(9):350?358. doi: 10.13386/j.issn1002-0306.2023050282.
LAN Meng, LI Jingfeng, LI Dongbing, et al. Anti-inflammatory Effects of Proteolytic Peptides from Different Ginseng Concoctions on LPS-induced RAW264.7 Cells[J]. Science and Technology of Food Industry, 2024, 45(9): 350?358. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2023050282.
Citation: LAN Meng, LI Jingfeng, LI Dongbing, et al. Anti-inflammatory Effects of Proteolytic Peptides from Different Ginseng Concoctions on LPS-induced RAW264.7 Cells[J]. Science and Technology of Food Industry, 2024, 45(9): 350?358. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2023050282.

人參不同炮制品蛋白酶解肽對LPS誘導RAW264.7細胞的抗炎作用研究

Anti-inflammatory Effects of Proteolytic Peptides from Different Ginseng Concoctions on LPS-induced RAW264.7 Cells

  • 摘要: 研究從人參炮制品生曬參、紅參、黑參中篩選出具有抗炎作用的蛋白酶解肽并進行對比分析。采用低溫浸提法從三種炮制品中提取人參不同炮制品蛋白,采用分步酶解法,利用堿性蛋白酶、中性蛋白酶和胃蛋白酶對三種人參蛋白進行酶解,得到BGP(黑參蛋白酶解肽)、RGP(紅參蛋白酶解肽)、SGP(生曬參蛋白酶解肽)三種酶解產物;使用超濾膜進行分離,分別得到不同分子量的超濾組分。通過建立脂多糖(LPS)誘導的RAW264.7炎癥模型確定了具有最強抗炎活性的組分。酶聯免疫法測定活性組分對RAW264.7細胞分泌一氧化氮(NO)、腫瘤壞死因子-α(TNF-α)、白細胞介素-1β((IL-1β)、白細胞介素-6(IL-6)的影響。分析三種蛋白酶解肽的氨基酸組成及含量。采用多元統計分析篩選出人參三種炮制品的差異性氨基酸并探究其與抑制RAW264.7細胞分泌細胞因子之間的關系。結果表明,人參三種炮制品蛋白酶解肽小于1 kDa組分較其他組分相比,對RAW264.7細胞增殖的作用最強且在質量濃度為50~200 μg/mL時可顯著抑制NO、TNF-α、IL-6、IL-1β的分泌(P<0.05)。質量濃度為200 μg/mL時,三種蛋白酶解肽給藥組對細胞因子釋放的抑制作用最強,且BGP-4對細胞因子釋放的抑制作用高于RGP-4和SGP-4,具有顯著差異(P<0.05)。三種蛋白酶解肽均含有17種氨基酸,但含量存在明顯差異。其中,苯丙氨酸含量最高且為三種人參炮制品差異氨基酸與抑制炎癥因子的分泌密切相關。本研究初步探討了炮制對人參抗炎活性的影響,并篩選出不同炮制品的差異性氨基酸,為人參炮制品的加工提供了參考依據。

     

    Abstract: The purpose of this study was to conduct a screening and comparative analysis of proteolytic peptides with anti-inflammatory properties derived from three different ginseng concoctions: Sundried ginseng, red ginseng and black ginseng. Ginseng proteins were extracted from three different types of ginseng products using a low-temperature leaching method. Subsequently, the extracted proteins underwent enzymatic digestion using alkaline protease, neutral protease, and pepsin through a stepwise enzyme digestion method. This process yielded three distinct enzyme digestion products, namely BGP (black ginseng proteolytic peptide), RGP (red ginseng proteolytic peptide), and SGP (sundried ginseng proteolytic peptide). The samples were subjected to separation using ultrafiltration membranes, resulting in the acquisition of ultrafiltration fractions with distinct molecular weights. Subsequently, the ultrafiltration fractions were further separated utilizing ultrafiltration membranes to obtain fractions with varying molecular weights. The fraction exhibiting the most potent anti-inflammatory activity was determined through the application of a lipopolysaccharide (LPS)-induced RAW264.7 inflammation model. The impact of the active fractions on the secretion of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-1β ((IL-1β), and interleukin-6 (IL-6) by RAW264.7 cells was assessed using enzyme immunoassay. The amino acid composition and content of the three proteolytic peptides were examined. Multivariate statistical analysis was employed to identify the distinct amino acids in the three ginseng concoctions and investigate their correlation with the inhibition of cytokine secretion by RAW264.7 cells. The findings of the study indicated that the proteolytic peptide fraction with a molecular weight of less than 1 kDa in the three ginseng products exhibited the most pronounced impact on the proliferation of RAW264.7 cells compared to the other fractions. Additionally, this fraction significantly suppressed the secretion of NO, TNF-α, IL-6, and IL-1β at concentrations ranging from 50~200 μg/mL (P<0.05). Notably, at a concentration of 200 μg/mL, the three groups receiving proteolytic peptide administration demonstrated the most potent inhibitory effect on cytokine release. Furthermore, the inhibitory effect of BGP-4 on cytokine release surpassed that of RGP-4 and SGP-4, exhibiting a statistically significant difference (P<0.05). All three peptides consisted of 17 amino acids, however, their compositions exhibited significant variations. Notably, phenylalanine exhibited the highest content, and the differential amino acids present in the three ginseng concoctions were closely associated with the inhibition of inflammatory factor secretion. This study represents an initial exploration into the impact of concoctions on the anti-inflammatory properties of ginseng, identifying distinct amino acids among different concoctions. These findings offer a valuable reference for the formulation of ginseng concoctions.

     

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