Preliminary Investigation On The Function Of TC0668 On The Regulation Of Cellular Glycometabolism By Chlamydia Murine

Background and Objective:On the one hand,Chlamydia as an obligate intracellular parasite relies on host cells to satisfy its energy and metabolic demands for growth and development;on the other hand,Chlamydial virulence factors have pathogenic effects on host cells to ensuring successful infection.Cell metabolism was affected by Chlamydia trachomatis（Ct）via the PI3K/AKT pathway,encouraging Ct to proliferate and grow more easily.The TC0668 virulence protein has been linked to the formation of mouse fallopian tube lesions induced by Chlamydia muridarum（Cm）,however the specific pathogenic mechanism is unknown.Cm,an effective alternate strain of Ct,may also impact pathogenicity by modulating host cell metabolism.We previously found that the associated differential protein of cells was mostly involved in metabolism and other processes after infected with Cm tc0668 single gene differential strains,and the PI3K activation and AKT phosphorylation were considerably different.As a result,we’ve attempted to discuss the impact on the cellular glycometabolism regulated by TC0668 of Cm,as well as the role of the PI3K/AKT pathway.This opens up new avenues for completely comprehending the function of TC0668 on Cm controlling cell metabolism and its mechanism,as well as delving deeper into the pathogenic mechanism of TC0668.Methods:1.The consumption level of glucose,the degree of mitochondrial Oxidative phosphorylation（OXPHOS）and aerobic glycolysis,and the extent of intracellular ATP were measured in He La cells and HUVECs infected by Cm TC0668^wt and Cm TC0668^mut strains at various times to investigate the impact of TC0668 on cellular glycometabolism.2.The activation of PI3K/AKT pathway in He La cells and HUVECs,was investigated to observe how Cm TC0668^wt and Cm TC0668^mut strains activated crucial molecules in the PI3K/AKT signaling pathway at different times.3.By inhibiting the activation of PI3K/AKT signaling pathway and analyzing the changes of cellular glycometabolism,we further elucidate the role of PI3K/AKT signaling pathway in the influence of TC0668protein on Cm regulation of cellular glycometabolism.Results:1.Compared with Cm TC0668^mut strains,the expression of mitochondrial Tricarboxylic acid cycle（TCA）-related protein Oxoglutarate Dehydrogenase（OGDH）was significantly increased after Cm TC0668^wt strains was infected with He La cells at 12 h or HUVEC cells at 24 h,but there was no significant difference in the expression level of mitochondrial electronic respiratory chain-related protein Cytochrome c oxidase subunit IV（COX IV）throughout the cycle.The levels consumption of glucose,the expression of Glucose Transporter 1（GLUT1）,and the extent of aerobic glycolysis and intracellular ATP were significantly difference in He La cells and HUVECs infected with Cm TC0668^wt strains compared with Cm TC0668^mut strains.2.Cm TC0668^wt strains remarkably improved PI3K activation and AKT phosphorylation compared to Cm TC0668^mut strains in the later stages of infection of He La cells and HUVECs.3.The expression of Glucose Transporter 1（GLUT1）was suppressed and the extent of intracellular ATP was boosted to a degree in He La cells and HUVECs,meanwhile,the consumption level of glucose was significantly suppressed in HUVECs but not in He La cells after Cm TC0668^wt strains infected cells that blocked the PI3K/AKT signaling pathway.Conclusions:1.The consumption degree of glucose,the extent of mitochondrial TCA cycle and aerobic glycolysis,and the level of intracellular ATP in host cells were influenced by TC0668 of Cm.2.The expression of GLUT1 and the levels of ATP in He La cells were impacts by TCO668 of Cm via activating the PI3K/AKT signaling pathway.3.The consumption degree of glucose and the level of intracellular ATP in HUVECs were influenced by TCO668 of Cm via stimulating the PI3K/AKT signaling pathway.