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Objective:To study the expression of DcR3 of myocardial tissue in diabetic mouse and normal rats and the impact of DcR3 recombinant protein to the expression of related molecules and myocardial cell apoptosis to discuss the action of DcR3 to myocardial cell apoptosis in Diabetic rats.Methods:Intraperitoneally injected streptozotocin one time to establish the model of Diabetic rats.Injected different doses of DcR3 recombinant protein to tail vein[1.2 mg/(rat? d),0.8 mg/(rat? d),0.4 mg/(rat? d)] 40 d. The expression of DcR3 mRNA, Fas mRNA and FasL mRNA of myocardial tissue was detected with RT-PCR;the expression of apoptosis related molecules Bcl-2 and Caspase-8 was analyzed with Western blot;the IL-1β, TNF-αand LFN-γof the blood was detected with double antibody sandwich ELISA;the percentage of myocardial cell apoptosis was observed with HE dyeing.Results:To compare the DcR3 treatment group with diabetic group,the expression DcR3 of myocardial tissue was high,the expression of Fas mRNA and FasL mRNA was descended.The Caspase-8 protein was ascended and the Bcl-2 protein was descended.The middle dose group was the most obvious.the IL-1β,TNF-αand IFN-γin the blood was descended differently in each DcR3 treatment group(P<0.05,P<0.01).The percentage of myocardial cell apoptosis was declined(P<0.05).Conclusion:DcR3 recombinant protein have the action of inhibiting the rats′myocardial cell apoptosis,the mechanism is related to competing with Fas,blocking-up FasL of inducing apoptosis, expressing DcR3 of myocardial cell,the descending of apoptosis related factors Caspase-8,the ascending of Bcl-2 and the reduction of cytokine levels.
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Objective:To study the effects of Anti-DR5 mAb on inducing synovial cells of adjuvant arthritis (AA) rats and its mechanisms.Methods:AA was induced by CFA in rats.The synovial cells of rats were separated and cultured in vitro system.The apoptosis induced by anti-DR5 mAb on synovial cells was measured by MTT analysis,DNA fragmentation and flow cytometry.Caspase-3 and Bcl-2 expression in synovial cells were detected by Western blot.The involvement of the apoptotic pathway was further proved by a caspase inhibition assay.Results:MTT result showed that Anti-DR5 mAb could inhibit synovial cells growth.And flow cytometry suggested that the cell death mode was apoptosis.The protein level of caspase-3 in synovial cells treated with anti-DR5 mAb was raised,while Bcl-2 level declined.When the caspase inhibitor was added to the synovial cells treated with anti-DR5 mAb,it was showed in a dose-dependence inhibition effect,indicating that anti-DR5 mAb inducing apoptosis might be through the caspase pathway.Conclusion:Anti-DR5 mAb could induce synovial cell apoptosis through caspase pathway.And this effect may be related to the protein level of Caspase-3 and Bcl-2.
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Objective:To study the cytotoxic effects on three glioma cell lines U343, U138, U373 induced by anti-human DR5/DR4 monoclonal antibodies(FMU1.5/FMU1.4) and the underlying mechanism.Methods:Expression of DR4/DR5 was quantitated by flow cytometry and DR/4DR5 mRNA detected by RT-PCR. Cytotoxicity exerted by FMU1.4/FMU1.5 on three cell lines was measured by MTT colorimetry and the induced apoptosis was determined by agarose gel electrophoresis, DNA ploidy analysis was studied by flow cytometry.Results:The expression of DR5 on U343 cells was higher and the expression of DR4 on U373 cells was lower. Cell line U343 was sensitive to FMU1.5 and in a dose dependent manner, but it was partially sensitive to FMU1.4; Cell line U138 was partially sensitive to FMU1.5 and resistant to FMU1.4; Cell line U373 was insensitive to two antibodies.Conclusion:Apoptosis induced by monoclonal antibodie FMU1.4/FMU1.5 vary among three cell lines. The underlying mechanism may be relevant to DR4/DR5 expression,the release of cytochrome C and FLIP.