四神丸对结肠炎小鼠结肠组织细胞能量代谢相关蛋白的调控作用OA

Objective To clarify the regulatory effect of Sishen Pill(四神丸,SSP)on cell energy metabolism-related proteins in the colon tissue of mice with dextran sulfate sodium salt(DSS)-induced colitis.Methods Forty-eight male C57BL/6 mice were randomly divided into four groups:Control group,Control+SSP group,DSS model group,and DSS+SSP group.The colitis model was established using 2.5%DSS solution.From the 7th day of the experiment,mice in the Control+SSP and DSS+SSP groups were gavaged with 2.5 g·kg-1·d-1 of SSP aqueous extract,while the other two groups received the same volume of sterile drinking water.Changes in body weight,fecal traits,and Disease Activity Index(DAI)were closely monitored;colon length and spleen weight were measured.Pathological damage of colon tissue was observed after HE staining.ELISA was used to detect the levels of energy metabolism-related nucleotides(ATP,ADP,etc.)in colon tissue.qRT-PCR was performed to assess the mRNA expression levels of ATPase-related genes.WB was employed to determine the protein expression levels of Glut1,Glut2,Glut3,Glut4,AMPK-α1,PGC-1α,and HIF-1α in colon tissue.Results Compared with the DSS group,the overall condition of mice in the DSS+SSP group was significantly improved:body weight increased significantly on the 17th day(P<0.05/P<0.01);DAI score showed a continuous downward trend from the 11th day(P<0.05/P<0.01);colon length was restored(P<0.01);spleen weight index and intestinal weight index decreased significantly(P<0.01);pathological score was significantly reduced(P<0.01);levels of AMP,ADP,ATP,and ATP ratio in colon tissue decreased signifi-cantly(P<0.05),while ADO level increased(P<0.05);mRNA expression levels of ATPase-related genes(ATP1B2,ATP1B3,ATP2B4,ATP2C1,ATP6V,ATP10A,ATP11A,ATP13A2,ATP13A3)increased to varying degrees(P<0.05/P<0.01),whereas mRNA expression levels of Notch3,ATP5E,and mTOR decreased(P<0.01);protein expression levels of Glut1,Glut2,Glut3,Glut4,and HIF-1α in colon tissue were significantly reduced(P<0.01),while protein expression levels of AMPK-α1 and PGC-1α increased significantly(P<0.01).Conclusion SSP alleviates glycolysis hyperactivity by inhibiting the HIF-1α/Glut axis,and simulta-neously activates the AMPK/PGC-1α pathway to promote mitochondrial biogenesis and ATP production.It reverses energy metabolism disorder and effectively ameliorates DSS-induced colitis in mice.