鹿茸血酶解肽对脂多糖诱导的H9c2大鼠心肌细胞损伤的保护作用

Protective Effect of the Enzymatic Peptide from Deer Antler Blood Against Lipopolysaccharide-induced Cardiomyocyte Damage in H9c2 Rats

摘要:

为研究鹿茸血酶解肽对脂多糖（LPS）诱导H9c2大鼠心肌细胞损伤的保护作用，本研究利用LPS刺激H9c2细胞建立损伤模型。选用卡托普利为阳性对照药。MTT法测定不同浓度（25、50、100、200、400 μg/mL）的鹿茸血酶解肽对H9c2细胞增殖抑制活性的影响；酶联免疫法测定细胞上清液中肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）含量。并分析鹿茸血酶解肽的氨基酸组成。结果表明，与模型组比较，不同浓度（25、50、100、200、400 μg/mL）的鹿茸血酶解肽均可显著抑制受损伤的H9c2增殖和TNF-α、IL-6、IL-1β的释放（p<0.05），模型组的TNF-α、IL-6、IL-1β释放量分别为：531.05、185.41、70.03 pg/mL，在200 μg/mL质量浓度下，鹿茸血酶解肽对上述三种炎症因子释放的抑制作用最显著（p<0.001），其释放量分别为：357.93、148.69、62.72 pg/mL。对LPS诱导H9c2细胞损伤具有保护作用的鹿茸血酶解肽中富含赖氨酸，含量占总氨基酸组成的17.81%。以上结果说明鹿茸血酶解肽可以抑制受损伤的H9c2细胞增殖,同时减少炎症因子TNF-α、IL-6、IL-1β的释放，通过抑制炎症因子的分泌来发挥其对LPS诱导的H9c2细胞损伤的保护作用。

Abstract:

In order to investigate the protective effect of the enzymatic peptide from deer antler blood aginst lipopolysaccharide (LPS)-induced H9c2 rat cardiomyocyte damage, this study used LPS to stimulate H9c2 cells to establish an injury model. Captopril was selected as the positive control drug. MTT method was used to determine the effects of different concentrations (25, 50, 100, 200, 400 μg/mL) of the enzymatic peptide from deer antler blood on H9c2 cell proliferation-inhibitory activity; Enzyme-linked immunoassay was used to determine the contents of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) in the cell supernatant. The amino acid composition of the enzymatic peptide from the antler blood was analyzed. The results showed that compared with the model group, different concentrations (25, 50, 100, 200, 400 μg/mL) of deer antler hemolyzed peptides could significantly inhibit the proliferation of damaged H9c2 and the release of TNF-α, IL-6 and IL-1β (p<0.05). The contents of the released TNF-α, IL-6 and IL-1β in the model group were 531.05, 185.41, and 70.03 (pg/mL), respectively. At a mass concentration of 200 μg/mL, the enzymatic peptide from the deer antler blood exhibited the greatest inhibitory effect on the release of the three above-mentioned inflammatory factors (p<0.001), with the released amounts as 357.93, 148.69, and 62.72 (pg/mL), respectively. Lys was rich in the enzymatic peptide from the antler blood that exhibited protection on the injured H9c2 cells induced by LPS, accounting for 17.81% of the total amino acids. These results indicate that the enzymolyzed peptide from deer antler blood can inhibit the proliferation of injured H9c2 cells, while reducing the release of inflammatory factors TNF-α, IL-6 and IL-1β, and exert its effect on LPS-induced H9c2 cells through inhibiting the secretion of inflammatory factors.