RESUMEN
OBJECTIVE@#: To investigate the effect of polysaccharides (LBPs) on TLR/NF-κB independent pathway and serum tumor necrosis factor (TNF-α) level in diabetic MyD88-knockout mice.@*METHODS@#: Diabetes was induced by feeding high-fat/high-sugar diet and injection of low-dose streptozotocin in MyD88-knockout mice. The diabetic mice were randomly divided into model group, positive control group and LBPs group. The expressions of TRAM, TRIF, TRAF6, RIP1 and TNF-α mRNA and proteins in mouse peritoneal macrophages were detected by real-time RT-PCR and Western blotting after LBPs treatment for 3 month. Serum TNF-α was determined with ELISA kit.@*RESULTS@#: Real time RT-PCR showed that compared with model group, the relative expressions of and mRNA in macrophages of LBPs group were significantly decreased and expression of was significantly increased (all 0.05).@*CONCLUSIONS@#: LBPs may not inhibit serum TNF-α level through TLR/NF-κB independent pathway.
Asunto(s)
Animales , Ratones , Diabetes Mellitus Experimental , Medicamentos Herbarios Chinos , Farmacología , Regulación de la Expresión Génica , Macrófagos Peritoneales , Ratones Noqueados , Factor 88 de Diferenciación Mieloide , Genética , FN-kappa B , Genética , Distribución Aleatoria , Transducción de Señal , Factor de Necrosis Tumoral alfa , Sangre , MetabolismoRESUMEN
Objective Quadratic regression universal rotary combination design was used to optimize the fluorescence labeling condition of lycium barbarum polysaccharide (LBP).Methods The fluorescence labeling condition of LBP presented, when the covalent coupling of LBP and tyramine was reacted with fluorescein isothiocyanate (FITC). Filter the best labeling condition via using quadratic regression universal rotary combination design experiment on the relationship of labeling efficiency among pH value of buffer solution, reaction time, temperature and the dose of tyramine.Results The regression equation was:Y=0.085 41 - 0.002 82X1 - 0.015 68X2 + 0.008 11X3 + 0.005 01X4 + 0.008 75X1X2 - 0.005 75X1X3 - 0.001 75X1X4 + 0.010 63X2X3 + 0.000 125X2X4 + 0.000 25X3X4 - 0.021 44X12 - 0.008 89X22 - 0.001 984X32 + 0.003 66 X42, and the variables fromX1 toX4 represented pH value, reaction time, temperature and dose of tyramine, respectively. The goodness of fitting of regression equation was statistically significant. The condition of labling LBP was optimized when the temperature was at 50℃, timing of labling at the fourth day and pH value was 8.5.Conclusion The condition of labling LBP was optimized with suitable temperature, pH value and extended timing.