| 基于肽键热振荡理论对僵蚕炮制前后体外抗帕金森活性的研究 |
| 投稿时间：2019-02-15 点此下载全文
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| 引用本文：李晶峰,王亚萍,边学峰,张辉,孙佳明.基于肽键热振荡理论对僵蚕炮制前后体外抗帕金森活性的研究[J].中国现代中药,2019,21(9):1229-1235 |
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| 基金项目:公益性行业科研专项（201507002） |
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| 中文摘要:目的：筛选僵蚕抗帕金森病作用活性组分，对比僵蚕传统热炮制加工前后活性，研究僵蚕天然寡肽与酶解寡肽抗帕金森病作用活性，并对比炮制前后天然与酶解寡肽收率，为僵蚕合理开发应用提供依据。方法：将僵蚕应用石油醚、乙酸乙酯、甲醇、水分别提取，得到僵蚕不同极性溶媒提取物，应用6-羟基多巴胺（6-OHDA）诱导的人神经母细胞瘤细胞（SH-SY5Y）细胞损伤模型对其全成分进行系统研究；进而将僵蚕水提取物经过超滤离心，分别得到>10 kDa、3~10 kDa、1~3 kDa、<1 kDa组分，再应用乙酰胆碱酯酶（AchE）抑制率及对6-OHDA诱导SH-SY5Y细胞损伤的保护作用进行活性组分筛选；基于肽键热振荡理论对炮制前后僵蚕不同组分活性进行对比研究；酶解僵蚕>10 kDa组分，对天然及酶解<1 kDa组分进行活性对比研究。结果：僵蚕水提取物较其他极性溶媒提取物活性强，水提取物中<1 kDa组分的活性最佳，因此确定僵蚕<1 kDa组分为活性组分；炒僵蚕<1 kDa组分寡肽含量高于生品；酶解<1 kDa组分的活性略低于天然寡肽，但酶解<1 kDa组分寡肽收率高于天然<1 kDa组分。结论：本文确定了<1 kDa组分为僵蚕体外抗帕金森的药效物质；基于肽键热振荡理论阐明了僵蚕经麸炒后，由于肽键的断裂，生成了更多的小分子寡肽类成分；由酶解得到收率更高的活性寡肽，解决了天然寡肽收率较低难于产业化的问题，为僵蚕开发应用提供了可靠的依据。 |
| 中文关键词:僵蚕 肽键热振荡 帕金森 炮制 |
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| Anti-Parkinson Activity of Bombyx Batryticatus in vitro before and after Processing Based on Thermal Oscillation Theory of Peptide Bond |
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| Abstract:Objective：To screen the active components of Bombyx Batryticatus against Parkinson′s disease，and contrast the activity of Bombyx Batryticatus before and after traditional hot processing，and to compare the anti-Parkinson′s disease activity and oligopeptide yield of natural oligopeptide and enzymatic oligopeptide of Bombyx Batryticatus，so as to provide reliable basis for rational development and application of Bombyx Batryticatus.Methods：Bombyx Batryticatus was extracted with petroleum ether，ethyl acetate，methanol and water respectively to obtain four different polar extracts.The whole components of the extracts were systematically studied using SH-SY5Y cell damage model induced by 6-hydroxydopamine（6-OHDA）.The water extracts of Bombyx Batryticatus were centrifuged by ultrafiltration to obtain components larger than 10，3-10 kDa，1-3 and less than 1 kDa.Then the inhibition rate of acetylcholinesterase and the protective effect of SH-SY5Y cells were screened.Based on the theory of thermal oscillation of peptide bond，the activity of different components of Bombyx Batryticatus before and after processing was compared，and the activity of natural and enzymatic hydrolysis components of Bombyx Batryticatus was compared.Result：The water extract of Bombyx Batryticatus had stronger activity than other polar solvents，and the component less than 1 kDa had the best activity，so it was determined that the component less than 1 kDa of Bombyx Batryticatus was the active component；the oligopeptide content of the component less than 1 kDa of bran fried Bombyx Batryticatus was higher than that of raw products；The activity of enzymatic oligopeptides less than 1 kDa fraction were slightly lower than that of natural oligopeptides，but the yield of oligopeptides with enzymatic hydrolysis less than 1 kDa fraction was higher than that of natural oligopeptides with enzymatic hydrolysis less than 1 kDa fraction.Conclusion：Components less than 1 kDa are identified as effective substances of Bombyx Batryticatus.The processing mechanism of Bombyx Batryticatus is clarified based on the theory of thermal oscillation of peptide bond，which indicates that after processing，more small molecular oligopeptides are produced due to the breakage of peptide bond.The active oligopeptides with higher yield are obtained by enzymatic hydrolysis.The problem that the yield of natural oligopeptides is low and difficult to industrialize is solved，which provides a reliable basis for the development and application of Bombyx Batryticatus. |
| keywords:Bombyx Batryticatus the theory of thermal oscillation of peptide bond Parkinson processing |
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