| Background : Glioma is the most common and difficult primary tumor in human nervous system tumors.The incidence rate of glioma(40%)is higher than that of intracranial tumors.Although in recent years,the treatment of glioma,such as surgery,radiotherapy,chemotherapy and other technologies have made great progress,but the overall treatment effect of glioma has not been significantly improved.Butyric acid is a kind of natural short chain fatty acid(SCFAs)which is produced by bacteria fermentation in the intestinal tract.It mainly exists in the form of sodium butyrate(Na B)in vivo.In recent years,it has been found that sodium butyrate can play an antibacterial role in the intestinal tract,not only promoting the regeneration and repair of intestinal epithelial cells,regulating the intestinal microecological balance,but also reducing body weight and improving insulin resistance.At the same time,sodium butyrate can also play a neuroprotective role through anti-inflammatory,anti-oxidation,inhibition of apoptosis and other ways.Moreover,sodium butyrate can inhibit the proliferation of a variety of tumor cells and promote the apoptosis of tumor cells.Mitochondrion is the main energy source of cells.It regulates the growth of cells by producing ATP,maintaining the stability of mitochondrial membrane potential(Δψm)and releasing apoptosis related factors.MPT(mitochondrial permeability transition),which is the key link in the mechanism of apoptosis,is mainly regulated by MPTP(mitochondrial permeability transition pore).When the opening of MPTP increases,it will lead to the decrease of mitochondrial membrane potential,the increase of mitochondrial membrane permeability(MMP),the decrease of ATP synthesis and the decrease of the coupling between respiratory chain and oxidativephosphorylation.ANT(adenine nuclear translocator),the ADP / ATP carrier,is the main transport protein of mitochondrial inner membrane,which was found in the early 1960 s.It is one of the most abundant proteins in mitochondrial inner membrane.The exchange of ADP and ATP in cells is controlled by proton electrochemical gradient,and an ADP is exchanged into an ATP molecule by electrophoretic mechanism.ANT plays an important role in the formation and opening of MPTP.It can exchange mitochondrial matrix ATP and cytoplasmic ADP,and control the production of intracellular ATP.ANT,as a member of mitochondrial transporter family,is the most abundant carrier in mitochondrial inner membrane,so it has attracted more and more attention in the transporter family.At present,there is no study on ANT and mitochondrial membrane permeability in the treatment of gliomas.Therefore,this study explored the regulatory mechanism of sodium butyrate on proliferation and apoptosis of U251 cells by studying the effect of sodium butyrate on mitochondrial membrane permeability.Purpose: In this study,sodium butyrate was used to treat U251 cells in vitro.Through the study of the effect of sodium butyrate on mitochondrial membrane permeability,explored the regulatory mechanism of sodium butyrate on the proliferation and apoptosis of glioma U251 cells.Contents and methods: In this study,we studied the effect of sodium butyrate on the proliferation and apoptosis of glioma U251 cells through mitochondrial membrane permeability,and explored the regulatory mechanism of sodium butyrate on the proliferation and apoptosis of glioma U251 cells.The glioma U251 cells were cultured in vitro and treated with sodium butyrate(0,1,2.5,5,10,20 mmol /L).CCK8 method was used to measure the proliferation activity of each group of cells,flow cytometry was used to detect the apoptosis level and mitochondrial membrane potential level of each group of cells,ATP detection kit was used to detect the expression level of ATP of each group of cells,seahorse instrument was used to detect the functional change of mitochondria of each group of cells,and Western blot was used to detect ANT2,Bcl-2,Bax,Caspase 3 and Cleaved-caspase 3 protein expression levels of each group of cells.Results: After treated U251 cells with sodium butyrate,the results showed that sodium butyrate significantly reduced the proliferation activity of U251 cells in a dose-dependent manner compared with the control group(P < 0.01).CCK-8 assayshowed the median inhibition concentration(IC50)of 48 h was 15.3 mmol / L.compared with the control group,sodium butyrate significantly increased the apoptosis rate of U251 cells in a dose-dependent manner(P < 0.05);sodium butyrate significantly reduced the ATP production level of U251 cells in a dose-dependent manner(P < 0.01);sodium butyrate significantly reduced the mitochondrial membrane potential of U251 cells,not only in a dose-dependent manner(P < 0.01),but also in a time-dependent manner(P < 0.05).sodium butyrate increased the function of mitochondria compared with the control group;compared with the control group and sodium butyrate group,CAT did not significantly increase the effect of sodium butyrate,but BA could significantly inhibit the effect of sodium butyrate.Compared with the control group,sodium butyrate reduced the protein expression level of Bcl2 and caspase 3(P < 0.05),but increased the protein expression level of ANT2,Bax and cleaved-caspase 3(P < 0.05).Conclusion: The results showed that sodium butyrate increased the mitochondrial membrane permeability of U251 cells by activating ANT2,decreased the mitochondrial membrane potential,decreased the production of ATP,increased apoptosis and decreased proliferation of U251 cells.Therefore,this study can show that sodium butyrate changes mitochondrial membrane permeability by regulating the function of ANT,and affects the proliferation and apoptosis of glioma.So sodium butyrate can play an important role in the prevention and treatment of glioma. |
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