The Expression Level Of Autophagic Gene And Intervention Of Autophagy By Metformin After Infected By Mycobacterium Tuberculosis

Mycobacterium tuberculosis(M.tb)is an intracellular bacteria,so the antibodies in the process of the immune response have play a role on extracellular M.tb infection and it had limited effect in M.tb permeated into cell.However,the innate immunesystem takes a significant role in resisting M.tb infection.Autophagy is component of innate immune system,whichparticipates in many of the innate immune signal control.Autophagy not only can degrade aging organelles and proteins,and it also removes bacteria and virus infection.Autophagy related proteins(ATG)are mutual cooperative,which induce the changes of cell membrane during the formation of autophagy.When the antigen presenting cells devoured the M.tb by a single membrane structure of the vesicles,andis modified by LC3 protein,then the autophagy proteins activation and modification to maturation.Autophagic bodies and lysosomes fuse with each other and degrade the devoured M.tb.The autophagy of cell not only has the ability to remove and kill the pathogen,it also takes a huge responsibility in the process of antigen presenting.The autophagy can promote MHC-II molecular bind with antigenic epitope of pathogen,and present antigen to CD4+Tcell.Besides,he autophagy can promote MHC-I molecular present antigen to CD8+T cell.We included 12 patients of first-treated tuberculosis treatment in Beijing Chest Hospital as case group and 12 newly recruited employees and students of Beijing Chest Hospital as the control group.In order to compare the level of autophagy gene we run the qPCR test.The research found that the mTOR gene level of health control group is about 2.76±1.56 times than case group and the Rheb gene level of health group is about 1.89±0.91 times than case group.It can be seen that the expression of these two genes is down regulated,which can promote the level of autophagy and resist M.tb infection.Besides,the research found that the BECN1 gene level of health control group is about 2.23±1.52 times than case group,which means the BECN1 gene leave of first-treated tuberculosis patients upgraded to inhibit the autophagy.We found that there are some diffident of autophagy gene expression between BECN1 and mTOR or they play the opposite role.This research also tests autophagy genes level of ATG5,ATG7,ATG12 and Rheb after infected by H37 Rv in 30 min,90min,4h,10 h and 24 h.It found that the autophagy gene of ATG5,ATG7 and ATG12 upgrade after infected 30 min.We also found that ATG5 and ATG7 gene expression up to hike after infected 10 h.However,there are no significant changes of Rheb gene in the period of infection between infectious groups and control groups and the different of Rheb genes in the two groups are within 2 fold.So we can conclude that the gene of Rheb has no changes after H37 Rv infection in the early time.We also used Metformin to intervene the THP-1 cells before it infected by BCG 4h ago.There were no significant changes of Rheb between Metformin groups and without Metformin groups.However,the mTOR gene was downgrade after the BCG infected 30 min.So the Metformin may inhibit the mTOR gene expression in the early period of BCG infection.In addition to these,the research also tests the amount of mRFP-GFP-LC3 B protein with or without the drug of Metformin.We found that the Metformin could assist the combination of autophagy bodies and lysosome,which may assist the autophagy of THP-1 cells infected BCG.