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Objective:To elucidate the key parameters associated with hydrogen peroxide induced oxidative stress and investigates the mechanism of trigonelline(TG)for reducing the H_2O_2induced toxicity in H9c2 cells.Methods:Cytotoxicity and antioxidant activity of TG was assessed by EZ-CYTOX kit.RNA extraction and cDNA synthesized according to the kit manufacture protocol.Apoptosis was measured by the Flowcytometry,general PCR and qPCR.Results:It was found that the TG significantly rescued the morphology of the H9c2 cells.Treatment of cells with TG attenuated H_2O_2 induced cell deaths and improved the antioxidant activity.In addition,TG regulated the apoptotic gene caspase-3,caspase-9 and anti-apoptotic gene Bcl-2.Bcl-XL during H_2O_2 induced oxidative stress in H9c2 cells.These results were comparable with quercetin treatment.For evident,flow cytometer results also confirmed the TG significantly reduced the H_2O_2 induced necrosis and apoptosis in H9c2 cells.However,further increment of TG concentration against H_2O_2 could induce the necrosis and apoptosis along with H_2O_2.Conclusions:It is suggested that less than 125μM of TG could protect the cells from H_2O_2 induced cell damage by down regulating the caspases and up regulating the Bcl-2 and Bcl-XL expression.Therefore,we suggest the trigonelline could be useful for treatment of oxidative stress mediated cardiovascular diseases in future.
Objective: To elucidate the key parameters associated with hydrogen peroxide induced oxidative stress and investigates the mechanism of trigonelline (TG) for reducing the H2O2induced toxicity in H9c2 cells. Methods: Cytotoxicity and antioxidant activity of TG was assessed by EZ-CYTOX kit. RNA Extraction and cDNA synthesis according to the kit manufacture protocol. Apoptosis was measured by the Flowcytometry, general PCR and qPCR. Results: It was found that the TG significantly rescued the morphology of the H9c2 cells. Treatment of cells with TG attenuated H_2O_2 induced cell deaths and improved the antioxidant activity.In addition, TG regulated the apoptotic gene caspase-3, caspase-9 and anti-apoptotic gene Bcl-2.Bcl-XL during H 2 O 2 induced oxidative stress in H9c2 cells.These results were comparable with quercetin treatment.For evident, flow cytometer results also confirmed the TG significantly reduced the H_2O_2 induced necrosis and apoptosis in H9c2 cells. Even, further increment of TG conc entration against H_2O_2 could induce the necrosis and apoptosis along with H_2O_2.Conclusions: It is suggested that less than 125μM of TG could protect the cells from H_2O_2 induced cell damage by down regulating the caspases and up regulating the Bcl-2 and Bcl-XL expression .Therefore, we suggest the trigonelline could be useful for treatment of oxidative stress mediated cardiovascular diseases in future.