Camargo Filho I, Cortez D A G, Ueda-Nakamura T, Nakamura C V, Dias Filho B P
Programa de Pós-graduação em Ciências Farmacêuticas, Brazil.
Phytomedicine. 2008 Mar;15(3):202-8. doi: 10.1016/j.phymed.2007.07.059. Epub 2007 Sep 24.
In this paper, we describe the purification of an antiviral peptide from seeds of Sorghum bicolor L. by a procedure that included gel filtration, ion exchange, and high-performance liquid chromatography (HPLC) in a reversed-phase column. Its molecular weight, determined by chromatographic mobility on the Shim-pack DIOL-150 gel permeation column in HPLC, was found to be 2000Da. The peptide designated 2kD peptide strongly inhibited the replication of herpes simplex virus type 1 (HSV-1), dose-dependently, at 40-90% of the control level, after incubation with 10-50 microM of the peptide, with EC(50) and EC(90) values of 6.25 and 15.25 microM, respectively. The IC(50) value of the 2kD peptide against Vero cells was 250 microM. Pre-incubation of HSV-1 with various concentrations of the 2kD peptide showed dose-dependent cytopathic effects (CPE) reduction patterns at concentrations from 6.25 to 50 microM. The presence of the 2kD peptide before HSV-1 infections showed moderate inhibition of virus-induced CPE as compared to during or after infections, with EC(50) values of 12.5, 6.25, and 6.25 microM, respectively. Similar results were observed when the 2kD peptide was assayed against bovine herpes virus (BHV), an enveloped virus like HSV-1. On the other hand, the 2kD peptide showed weak activity against poliovirus type 1, a non-enveloped virus. Taken together, these results indicate that the 2kD peptide was able not only to inhibit the initiation and the spread of infection, but also had an in vitro prophylactic effect against HSV-1 infection.
在本文中,我们描述了从高粱种子中纯化一种抗病毒肽的过程,该过程包括凝胶过滤、离子交换以及在反相柱上进行的高效液相色谱(HPLC)。通过HPLC在Shim-pack DIOL-150凝胶渗透柱上的色谱迁移率测定,其分子量为2000Da。命名为2kD肽的该肽在与10 - 50μM的肽孵育后,以剂量依赖方式强烈抑制单纯疱疹病毒1型(HSV-1)的复制,抑制水平为对照水平的40 - 90%,其半数有效浓度(EC(50))和90%有效浓度(EC(90))值分别为6.25和15.25μM。2kD肽对Vero细胞的半数抑制浓度(IC(50))值为250μM。用不同浓度的2kD肽对HSV-1进行预孵育,在6.25至50μM的浓度范围内呈现出剂量依赖性的细胞病变效应(CPE)降低模式。与感染期间或感染后相比,在HSV-1感染前存在2kD肽显示出对病毒诱导的CPE的中度抑制,其EC(50)值分别为12.5、6.25和6.25μM。当针对牛疱疹病毒(BHV,一种与HSV-1类似的包膜病毒)检测2kD肽时,观察到了类似的结果。另一方面,2kD肽对1型脊髓灰质炎病毒(一种非包膜病毒)显示出较弱的活性。综上所述,这些结果表明2kD肽不仅能够抑制感染的起始和传播,而且对HSV-1感染具有体外预防作用。