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目的:用高速逆流色谱法从青果氯仿萃取液中分离出不同组分,考察其抗人类免疫缺陷病毒(HIV-1)的活性并初步探讨其作用机制。方法:青果的氯仿萃取液用高速逆流色谱法分离出各组分;利用HIV-1假病毒检测各组分抑制HIV-1感染的活性;XTT比色法检测各组分对细胞的毒性;采用酶联免疫吸附测定法(ELISA)研究各组分体外抗病毒活性的机理。结果:青果氯仿萃取液能抑制HIV-1JRFL及HIV-1HXB2假病毒感染活性;用高速逆流色谱法从青果氯仿萃取液中分离出3个组分:组分Ⅰ,组分Ⅱ,组分Ⅴ,其中组分Ⅱ纯度达90%以上,3个组分均能抑制HIV-1JRFL及HIV-1HXB2假病毒感染的活性,其中组分Ⅰ和组分Ⅴ能抑制HIV-1 gp41六螺旋的形成,其半数抑制浓度分别为(31.42±3.00)、(94.10±14.87)μg·ml-1。结论:用高速逆流色谱法从青果中分离了具有抗HIV-1感染活性的不同组分。
OBJECTIVE: To separate and identify different components of chloroform extract by high-speed countercurrent chromatography and investigate its anti-HIV-1 activity and to explore its mechanism. Methods: The constituents were isolated by high speed countercurrent chromatography. The HIV-1 infection was detected by HIV-1 pseudovirus. The cytotoxicity of each component to the cells was detected by XTT assay. Enzyme-linked immunosorbent assay (ELISA) to study the mechanism of antiviral activity of each component in vitro. Results: Chloroform extract could inhibit the HIV-1JRFL and HIV-1 HXB2 pseudovirus infection. High-speed countercurrent chromatography was used to separate three components from the chloroform extract: component I, component II, component V, The purity of component Ⅱ was more than 90%. The three components all could inhibit the HIV-1JRFL and HIV-1HXB2 pseudovirus infection. Components Ⅰ and Ⅴ could inhibit the formation of HIV-1 gp41 six-helix, The median inhibitory concentrations were (31.42 ± 3.00) and (94.10 ± 14.87) μg · ml-1, respectively. Conclusions: Different components with anti-HIV-1 infection activity were isolated from the fruits using high-speed countercurrent chromatography.