Research On The Role Of ADPN/FGF9 Pathway In Chronic Stress-induced Depression And Its Drug Treatment

Objective:The aim of this study is to explore the role of ADPN/FGF9 pathway in chronic stress induced depression and its pharmacological treatment through clinical studies and animal experiments.Methods:（1）Serum levels of ADPN/FGF9 in patients with depression:10 patients with first episode depression（depression group）and 10 healthy controls（control group）were enrolled.The serum levels of adiponectin and FGF9 were measured in all the cases.（2）The effect of ADPN/FGF9 pathway on depression model in mice:（1）The depressive behavioral research about different months adiponectin gene knockout mice:The depressive behavioral evaluation（sucrose preference,novelty suppressed feeding,forced swimming）respectively for 3 months,9 months,12 months of age male wild type（WT）mice and Adipo^-/-mice,After behavioral detection,ADPN and FGF9 contents in serum and hippocampal tissue homogenate were detected by ELISA,as well as the hippocampal tissue for histopathological examination.（2）The effects of ADPN or anti-FGF9 on depressed ICR mice by injection in the Lateral Ventricle:40 male ICR mice for 4 weeks,18²2g,were taken 10 mice as the normal control group,other mice after 4 weeks of continuous processing of CUMS and were randomly divided into 3 groups:a CUMS group,a ADPN group,a anti-FGF9 group And then,the mice of each group were injected into the lateral ventricle.The normal control group and the CUMS group were injected with the phosphate buffer solution（PBS）in the lateral ventricle,and the ADPN group and the anti-FGF9 group were injected with recombinant ADPN and anti-FGF9 in the lateral ventricle,respectively.Behavior detection（sucrose preference,novelty suppressed feeding and forced swimming）were test after a night recovery.The contents of ADPN and FGF9 in serum and hippocampus were detected by ELISA.Hippocampal tissues were examined by routine histopathology.（3）The Effects of anti-FGF9 on depressed Adipo^-/-mice by injection in the Lateral Ventricle:30 male Adipo^-/-mice for 3mouths,except for the Adipo^-/-mice in blank group,other mice after 4 weeks of continuous processing of CUMS and were randomly divided into 2 groups:a Adipo^-/-model group,a anti-FGF9 group.The rats in each group were injected into the lateral ventricle after grouping,the Adipo^-/-control group and the Adipo^-/-model group were injected with PBS in the lateral ventricle,and the Adipo^-/-anti-FGF9 group was injected with anti-FGF9 in the lateral ventricle.Behavior detection（sucrose preference,novelty suppressed feeding and forced swimming）were test after a night recovery.After the behavioral examination,hippocampal tissues were examined by routine histopathology.（3）The effect and mechanism of natural drug Carnosic acid（CA）on CUMS depression mice:60 male ICR mice for 4 weeks,18²2g,were randomly divided into 6 groups by weight:normal control group,CUMS group,fluoxetine group,low（25mg/kg）,medium（50mg/kg）,high（100mg/kg）dose groups of carnosic acid,all groups had 10 mice.Except the normal control group,CUMS intervention was performed for 28 days in the other groups,and the appropriate dose of medication was given at the same time after the 14^th CUMS,the normal control group and CUMS model group were given the same volume of distilled water daily,and the volume of the drug was 20ml/kg.After 14 days of administration,all the groups of animals were given the sucrose preference test,open field test,novelty suppressed feeding test,and forced swimming test.After that,ELISA method was used to detect the content of ADPN and FGF9 in serum and hippocampus,and histopathological examination of the hippocampus was performed at the sane time.Results:（1）Serum levels of ADPN/FGF9 in patients with depression:There was no significance in anthropometric parameters between the two groups（P>0.05）.The serum levels of ADPN in the depression group were significantly lower than those in the control group（P<0.01）and the content of FGF9 was significantly higher than those in the control group（P<0.05）.（2）The effect of ADPN/FGF9 pathway on depression model in mice:（1）the depressive behavioral research about different months adiponectin gene knockout mice:a.Behavioral test:compared with the same month old mice,3 month old,9 month old and 12 month old Adipo^-/-in sucrose preference index and total water intake,intake of sugar content and feeding latency were not statistically significant（P>0.05）;in the forced swimming test in mice,only 12 month old Adipo^-/-immobility time was significantly prolonged（P<0.05）.b.The contents of FGF9and ADPN in serum and hippocampal tissues:compared with the sane month old mice,3month old Adipo^-/-mice serum FGF9 content had no significant difference（P>0.05）;increased FGF9 content of 9 month old and 12 month old Adipo^-/-in serum,FGF2 content decreased significantly（P<0.05）;the content of ADPN is lower than that of WT mice Adipo^-/-mice serum in（P<0.05）;there was no significantly difference in hippocampal homogenate level of FGF9 and ADPN between all month Adipo^-/-mice（P>0.05）.c.Hippocampal tissues were examined by routine histopathology.In WT mice and Adipo^-/-mice with 3 month old or 9month old,the hippocampal cells were arranged tightly,and the vertebral cells were normal.The hippocampal cells in 12 month old Adipo^-/-mice were arranged in a sparse disorder,and some vertebral body cells were necrotic.（2）The effects of ADPN or anti-FGF9 on depressed ICR mice by injection in the Lateral Ventricle:a.Weight:Compared with the normal control group,the weight of CUMS group was significantly reduced（P<0.01）in the 7th day,14 days,21 days and 28 days.b.Behavior test:compared with normal control group,CUMS group were significantly decreased sucrose preference index（P<0.01）,feeding latency and immobility time was significantly increased（P<0.01）;compared with CUMS group,ADPN group significantly increased the sucrose preference index（P<0.05）,anti-FGF9 group showed an upward trend;ADPN group and anti-FGF9 group were significantly increased in feeding latency and immobility time（P<0.05 or P<0.01）;c.The FGF9 and ADPN contents in serum and hippocampal tissues:Compared with the normal control group,CUMS group in serum and hippocampus homogenate levels of ADPN were significantly decreased（P<0.01）,the content of FGF9 increased significantly（P<0.05 or P<0.01）;compared with CUMS group,the content of FGF9 in serum and hippocampal homogenates of both ADPN group and anti-FGF9group was significantly decreased（P<0.05 or P<0.01）.),the content of ADPN in hippocampal homogenate in both ADPN group and anti-FGF9 group showed an upward trend.d.The routine histopathological examination of hippocampal tissue:The ADPN group and the anti-FGF9 group all slightly improved the arrangement of hippocampal cells and the necrosis of the vertebral body.e.The hippocampus immunohistochemical examination:Compared with normal control group,the expression of FGFR3 decreased significant in the CUMS model group（P<0.01）,and compared with model group,The ADPN group and the anti-FGF9 group all hippocampus FGFR3 expression quantity increased significantly（P<0.05）.（3）The Effects of anti-FGF9 on depressed Adipo^-/-mice by injection in the Lateral Ventricle:a.Weight:Compared with the normal control group,the weight of adipo^-/-model group was significantly reduced after the model in the 14th day,21 days and 28 days（P<0.01）.b.Behavioral test:Compared with the normal control group,Adipo^-/-model group,sucrose preference index decreased significantly（P<0.01）,feeding latency and immobility time was significantly increased（P<0.01）;compared with CUMS group,FGF9 antibody group immobility time were significantly increased（P<0.05）,sucrose preference index and feeding latency increased;c.Routine histopathological examination of hippocampal tissue:The Adipo^-/-anti-FGF9 group slightly improved the arrangement of hippocampal tissue cells and the necrosis of the vertebral body;d.The hippocampus immunohistochemical examination:Compared with normal control group,the expression of FGFR3 decreased significant in the CUMS model group（P<0.01）,and compared with model group,the Adipo^-/-anti-FGF9 group hippocampus FGFR3expression quantity increased significantly（P<0.05）.（3）The effect and mechanism of natural drug Carnosic acid（CA）on CUMS depression mice:a.weight:Compared with the normal group,the weight of the model group decreased significantly（P<0.01）.Compared with the model group,the low dose of CA（25mg/kg）,middle dose of CA（50mg/kg）and high dose of CA（100mg/kg）could significantly increase the body weight of mice（P<0.05 or P<0.01）.b.Behavioral test:The medium dose of CA group significantly increased the number of mice spontaneous activity and sugar water preference index（P<0.05）,and the medium and high dose of CA group could significantly reduce the incubation latency and immobility time（P<0.05 or P<0.01）.c.The FGF2、FGF9 and ADPN contents in serum and hippocampal tissues:The middle and high dose of dose,the content of FGF9 in serum and hippocampus（P<0.05）was significantly reduced,and the content of FGF2and ADPN in serum and hippocampus was increased（P<0.05）.d.Routine histopathological examination of hippocampal tissue:The high dose of CA could improve the arrangement of hippocampus and the necrosis of the vertebral body.Conclusion:The serum levels of ADPN and FGF9 in patients with first episode depression are usually abnormal;The ADPN/FGF9 pathway may be closely related to cums-induced depression;The natural drug CA can significantly improve the depression status of CUMS mice,and its anti-depression effect may be related to the regulation of ADPN/FGF9 pathway.