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Vif, one of the six accessory genes expressed by HIV-1, is essential for the productive infection of natural target cells. Previously we suggested that Vif acts as a regulator of the viral protease (PR): It prevents the autoprocessing of Gag and Gag-Pol precursors until virus assembly, and it may control the PR activity in the preintegration complex at the early stage of infection. It was demonstrated before that Vif, and specifically the 98 amino acid stretch residing at the N'-terminal part of Vif (N'-Vif), inhibits both the autoprocessing of truncated Gag-Pol polyproteins in bacterial cells and the hydrolysis of synthetic peptides by PR in cell-free systems. Linear synthetic peptides derived from N'-Vif specifically inhibit and bind HIV-1 PR in vitro, and arrest virus production in tissue culture. Peptide mapping of N'-Vif revealed that Vif88-98 is the most potent PR inhibitor. Here we report that this peptide inhibits both HIV-1 and HIV-2, but not ASLV proteases in vitro. Vif88-98 retains its inhibitory effect against drug-resistant HIV-1 PR variants, isolated from patients undergoing long-term treatment with anti-PR drugs. Variants of HIV protease bearing the mutation G48V are resistant to inhibition by this Vif-derived peptide, as shown by in vitro assays. In agreement with the in vitro experiments, Vif88-98 has no effect on the production of infectious particles in cells infected with a G48V mutated virus.  相似文献   
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摘要 目的:探讨甜橙黄酮通过AMP依赖的蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路对呼吸道合胞病毒(RSV)感染大鼠肺组织损伤的改善作用。方法:将30只雄性Wistar大鼠分为对照组、模型组、甜橙黄酮低剂量组(14 mg/kg)、甜橙黄酮中剂量组(28 mg/kg)、甜橙黄酮高剂量组(56 mg/kg)、甜橙黄酮高剂量+AICAR组(56 mg/kg+500 mg/kg的AMPK激活剂AICAR),计算大鼠肺指数。实时荧光定量聚合酶链式反应(qRT-PCR)法测定肺组织RSV病毒载量,酶联免疫吸附(ELISA)法测定大鼠肺泡灌洗液炎症因子水平,HE染色测定肺组织病理,蛋白免疫印迹(Western blot)法测定AMPK/mTOR通路蛋白表达。结果:与对照组相比,模型组大鼠肺组织存在炎性浸润,排列松散,肺指数、肺组织RSV病毒载量、肺泡灌洗液白细胞介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)水平、炎症评分、p-AMPK/AMPK、mTORC1蛋白表达升高(P<0.05);与模型组相比,甜橙黄酮低剂量组、甜橙黄酮中剂量组、甜橙黄酮高剂量组大鼠肺组织损伤减轻,大鼠肺指数、肺组织RSV病毒载量、肺泡灌洗液IL-1β、IL-6、TNF-α水平、炎症评分、p-AMPK/AMPK、mTORC1蛋白表达降低(P<0.05);与甜橙黄酮高剂量组相比,甜橙黄酮高剂量+AICAR组大鼠肺组织损伤加重,肺指数、肺组织RSV病毒载量、肺泡灌洗液IL-1β、IL-6、TNF-α水平、炎症评分、p-AMPK/AMPK、MTORC1蛋白表达升高(P<0.05)。结论:甜橙黄酮可能通过抑制AMPK/mTOR信号通路发挥对RSV诱导的大鼠抗炎、抗病毒、抗肺部损伤作用。  相似文献   
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Orthosiphon aristatus is a traditional folk medicine extensively used in Southeast Asia because of its various pharmacological effects, including antioxidant, antitumor, and hypoglycemic activities. Orthosiphon extracts have been found to be cytotoxic to hepatocellular carcinoma (HCC) cells, which is attributed to their phytochemical content. However, the mechanism of action underlying the cytotoxic effects remains unclear. Hence, the present study investigated the effect of Sinensetin purified from O. aristatus on HCC in vitro. Sinensetin was isolated from O. aristatus leaves and the chemical structure was confirmed by ultra violet (UV)-vis, infrared (IR), nuclear magnetic resonance (NMR), and electrospray ionization mass spectrometry (ESI-MS). The results revealed that 24-h treatment with the purified compound markedly inhibited the survival of HepG2 cells, with IC50 of 39.93 ± 1.10 μg/mL. HepG2 cells treated with the IC50 of Sinensetin showed characteristic morphological changes, as determined by PI and AO/Etbr dual staining, including DNA fragmentation, thus confirming the apoptosis induction. Sinensetin induced cell cycle arrest at G0/G1 phase, and the data were substantiated by flow cytometry. Furthermore, Sinensetin modulated key signaling molecules; anti-apoptotic Bcl-xL was down-regulated, whereas the expressions of tumor suppressors TRAIL and PTEN were up-regulated. We conclude that Sinensetin can be effective against HCC.  相似文献   
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