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Objective:To determine the effects of TPS on peripheral blood Tregs in sepsis mouse induced by burn plus P.aeruginosa infection.Methods: The experimental mice were separated into five groups randomly ,including sham burn group ,burn plus P.aeruginosa infection group ,burn plus P.aeruginosa infection with TPS (50,100,200 mg/kg) treatment group.Peripheral blood Tregs were isolated with Magnetic Microbeads and cultured in vitro from the day after burn (PBD0) to 4 days after burn(PBD4).IL-10, IFN-γ,IL-4 levels in Tregs culture supernatants were determined by sandwich enzyme-linked immunosorbent assays ( ELISA ) . Purification of CD4+CD25high Tregs and CD4+T cells in C57BL/6 mice were administrated by magnetic beads sorting .Tregs and CD4+T cells were cultured in vitro after joining TPS to without TPS cells as a control .The phenotypes of Tregs were analyzed by flow cytometry , and cytokines were measured by ELISA .Results:Vis-a-vis the results of the untreated group ,TPS could markedly decrease IL-4 and IL-10 secretion level and significantly increase the secretion of IFN-γ,and the secretion of IL-10 level and concentration of TPS dose effect.Vis-a-vis the results of the untreated group ,in vitro experiment ,without stimulation of TPS ,CD4+T cell proliferation and IFN-γwere significantly reduced ( P<0.05 ) and IL-4 levels increased significantly;CD4+T cell proliferation and IFN-γwere significantly increased and IL-4 levels were significantly reduced in the group of TPS with antibody-1;there was no significant difference in CD 4+T cell proliferation and the levels of IFN-γand IL-4 in the group of TPS with antibody-2.Conclusion:TPS could inhibit the abnormal ac-tivities of CD4+CD25highTregs in burn with P.aeruginosa infection mice,at least in part via inhibiting IL-10 secretion,and trigger a shift of Th2 to Th1 with activation of CD4+T cells in burn with P.aeruginosa infection mice.
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Objective:To optimize the ultrasonic-assisted extraction technology of polysaccharide from Tremella by Box-Benhnken response surface methodology .Methods:The single-factor extraction tests and response surface analysis were employed to optimize the technology conditions , including ultrasonic power , ultrasonic time and liquid/solid ratio.Results:The experiment results indicated that the optimal extraction conditions were ultrasonic power of 360 W, ultrasonic time of 23 min and liquid/solid ratio of 45 ∶1 ( ml· g-1 ) . Under the conditions , 6 batches of polysaccharide from Tremella were extracted .The average extraction rate of Tremella polysaccharides was 20.4%±1.8%(n=6).Conclusion: The ultrasonic-assisted extraction process of Tremella polysaccharide using Box-Behnken response surface methodology is simple with good predictability .
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Objective: To observe the anti-apoptosis effect of tremella polysaccharides (TP) on cardiomyocytes by in vitro and in vivo experiments. Methods: TP was extracted by dehydrated alcohol and the TP purity was measured by gas chromatography. The cultured cardiomyocytes of the neonatal rats were randomly assigned to normal group (Group A), apoptosis-induced group (Group B) and TP-pretreatment group (Group T). The morphology and uptake rate of trypan blue were studied after 72 h. The apoptosis index of cardiomyocyte was measured by flow cytometry. In the in vivo study, 100 ICR mice were randomly assigned to 5 groups after paired by body weight as following: Group N (negative control group, peritoneally injected by saline daily); Group C (positive control group, peritoneally injected by D-galactose and saline daily); Group L, M, and H (peritoneally injected by D-galactose and intragastric administered with TP at 100, 200, 400 mg/day, respectively). Mice were sacrificed for histology and biochemistry studies after 8 weeks. Results: Data of cultured cardiomyocytes were as follows: Uptake rate of trypan blue: B>C>A (PC> A (P<0.01). In the animal studies there were no obvious difference in myocardium pathology between groups. The order of the apoptosis indices of myocardium was H
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@#myocytes.In aging mice induced by D-galactose experiment.TP has anti-apoptosis and anti-oxidation effect in a dose-dependant manner.
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ificantly lower than in both middle and low group(P＜0.05).Conclusion TP may improve the antioxidant ability of experimental aging mice.
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Objective:To observe the protective effect of tremella polysaccharides (TP) on mouse aging model induced by D-galactose. Methods: Forty male ICR mice were divided into 5 groups (Group A: treated with 800 mg/kg TP; Group B: treated by 400 mg/kg TP; Group C: treated by 200 mg/kg TP; Group D: aging control; Group E: normal control). Except for Group E,other groups were injected with D-galactose [120 mg/(kg?d)] for 60 d to establish subacute aging model. Group A,B and C were treated with 800,400 and 200 mg/kg TP respectively via intragastric (i.g.) administration. Sixty days later,the spleen,heart, kidney and brain were collected to determine MDA, LP, SOD, GSH-Px and contents of hydroxyproline by colorimetry. Expression of P21 in the kidney were detected by immunohischemical technique. Proliferation and transformation of spleen lymphocytes induced by ConA were determined by MTT method. Results: Antioxidants SOD and GSH-Px activity of TP treated groups were higher than aging control group (P