| Objective:Guillain-Barre syndrome(GBS)is an autoimmune mediated peripheral neuropathy.Acute motor axonal neuropathy(AMAN)is one of the common subtypes of GBS.The pathogenic antibodies are anti-ganglioside(mainly GM1 and GD1a)antibodies.Recent studies have shown that in addition to directly mediating axonal damage,antiganglioside antibodies can also inhibit axonal regeneration after neuronal damage.It has been reported that Adenosine monophosphate-activated protein kinase(AMPK),as a receptor of cellular energy metabolism,plays a key role in the growth and regeneration of neuronal axons by regulating the level of cell energy metabolism.In this study,the AMAN cell model was established by interfering with NG108-15 cells with anti-GM1 IgG antibody.The purpose of this study was to investigate the effect of anti-GM1 IgG on AMPK expression and axonal growth in NG108-15 cells,so as to provide ideas for exploring a new therapeutic regimen for AMAN.Method:This study used anti GM1 IgG intervention to establish a cell model of AMAN in NG108-15 cells.Firstly,the expression of GM1 on the surface of NG108-15 cells was verified using fluorescein labeled cholera toxin;cells were divided into AMAN model group,blank group,and control group(with normal IgG added)based on whether anti GM1 IgG was added.The effect of anti GM1 IgG antibody on axonal growth of NG108-15 cells was studied by measuring axonal length and growth cone angle;exploring the mechanism of action by immunofluorescence detection of AMPK expression and its phosphorylation level in each group of cells,as well as the expression of phosphorylated mammalian target of rapamycin(p-mTOR).Results:1.The immunofluorescence results showed that GM1 was successfully expressed on the cell membrane of NG108-15 cells.2.Anti GM1 IgG significantly inhibited the axonal growth of NG108-15 cells in the model group:the axonal length in the model group was significantly shorter than that in the blank group and control group(both P<0.001);the disintegration test of axonal growth cones showed that the growth cone angle of the model group was smaller than that of the blank group(P<0.001),and there was no statistical difference compared to the control group(P=0.5497).3.Immunofluorescence analysis showed that the expression levels of AMPK and p-AMPK in the model group cells were significantly higher than those in the blank group and control group(P=0.005,P<0.001,respectively);meanwhile,the expression level of p-mTOR in the model group was higher than that in the blank group(P<0.001),while there was no statistically significant difference in the expression level of p-mTOR between the model group and the control group(P=0.2036).Conclusion:Anti GM1 IgG may inhibit axonal growth by promoting the expression level of AMPK in NG108-15 cells. |
