ABSTRACT
Aim To establish and optimize a method for screening HIV-1 integrase 3′-processing inhibitor. Methods Fluorescence resonance energy transfer ( FRET) was used to create an assay for screening in-tegrase 3′-processing inhibitors; wavelength was de-fined by DNaseⅠ; factors affecting IN activity were optimized, including buffer composition, substrate con-centration, enzyme concentration, metal ion concentra-tion. Results Integrase 3′-processing optimizing reac-tion conditions were buffer 1 , 500 nmol · L-1 sub-strate, 1 μmol·L-1 integrase, 20mmol·L-1 magne-sium ion. Positive drug raltegravir and myricetin could effectively inhibit integrase 3′-processing activity using this assay. Two integrase 3′-processing inhibitors were screened by this method. Conclusion The method for screening HIV-1 integrase 3′-processing inhibitor is successfully established and optimized.
ABSTRACT
Objective:To explore the anti-inflammatory effects and the underlying mechanisms of ethanol extract of Ageratum cony-zoides. L. from Guangxi. Methods:The auricle edema model was induced by dimethylbenzene in the mice and the paw edema model was induced by carrageenan respectively in the mice and rats to study the anti-inflammatory effects of ethanol extract of Ageratum cony-zoides. L. from Guangxi. The content of malondiadehyde (MDA) and proateglandin E2 (PGE2), and the activity of superoxide dis-mutase( SOD) in the mouse edema paw was measured. The contents of tumour necrosisfactor-α ( TNF-α) , interleukin-1β ( IL-1β) and interleukin-6(IL-6) in the rat serum were detected as well. Results:Compared with the model control group, the ethanol extracts of Ageratum conyzoides. L. from Guangxi could remarkably inhibit auricle edema in the mice and paw edema in the mice and rats( P<0. 05 or P<0. 01), the inhibition ratio for high, medium and low dosage group(6. 0, 3. 0, 1. 5 g·kg-1)was 29. 24%,16. 42% and 11. 21% in the auricle edema mice and 28. 66%,18. 79% and 13. 13% in the paw edema mice , respectively. It could remarkably re-duce MDA and PGE2 content and enhance the activity of SOD in the mouse inflammatory tissue(P<0. 05 or P<0. 01). In the paw e-dema rats, the inhibition ratio for high, medium and low dosage group(4. 5,2. 3, 1. 2 g·kg-1)was 43. 69%, 36. 01% and 23. 29%at the 3rd h, respectively , and it also could remarkably reduce serum TNF-α, IL-1βand IL-6 content(P<0. 05 or P<0. 01). Con-clusion:The ethanol extracts of Ageratum conyzoides. L. from Guangxi show significantly anti-inflammatory effects, and the mecha-nisms may be related to the ability of scavenging oxygen free radicals and reducing the release of inflammatory cytokines and proinflam-matory cytokines.