Antidepressant Effect And Mechanism Of Gardenia Blue

Objective : To investigate the degree of polymerization of geniposide with different amino acids to produce gardenia blue,and determine the best complexing conditions.The depression model of rats was established by CUMS combined with orphan rearing.Through the changes of behavior,monoamine neurotransmitter and hippocampal tissue,gardenia blue was selected to further analyze the structure and metabolic characteristics of intestinal flora in each group,and to explore the antidepressant effect and mechanism of different gardenia blue;In order to investigate the effects of GAD and GHD on the damage and repair of neurons,a cell injury model of PC12 cells induced by glutamate(Glu)was established.The regulation of LDH,ROS,SOD and GSH was analyzed.The transcription of tau,BDNF and CREB was analyzed.Objective to explore the protective effect and mechanism of GAD and GHD on neuron cells,and to provide basis for the development of gardenia blue for antidepressant drugs or food therapy products.Methods: 1.Study on the complexation of genipin with different amino acids Genipin with different molar ratio and 7 kinds of amino acids were added to PBS buffer solution,heated and stirred at 80 ℃ for 4 h to obtain gardenia blue complex solution.The content of genipin in different gardenia blue solutions was determined by HPLC to investigate whether the complex was complete.2.The antidepressant effect of gardenia blue on cums rats(1)Establishment of a rat model of depression and its behavioral study The depression model of rats was established by isolated rearing and chronic unpredictable mild stress(cums).The behavioral indexes of control group,model group,fluoxetine group and different gardenia blue groups were investigated by sugar consumption test,open field test,forced swimming test and tail suspension test.(2)Effects of different gardenia blue on hippocampus and monoamine neurotransmitters in depression rats Paraffin embedding and he staining were used to observe the pathological changes of hippocampus.The contents of 5-HT and DA in brain tissue and serum of rats were detected by ELISA kit,and the regulatory effects of different gardenia blue on them were analyzed,so as to explore its mechanism.(3)Regulatory effects of GAD and GHD on intestinal flora in depression rats Before the end of the experiment,the faeces of rats in each group were collected,and the carbon source metabolism of faeces of rats in each group was investigated by using six kinds of selective media and Biolog eco plate.The awcd value,diversity index of intestinal flora and carbon source metabolism characteristics of intestinal flora of rats in each group were analyzed.The difference and similarity of intestinal flora in each group were analyzed by principal component analysis and cluster analysis to further explore its mechanism.3.The protective effects of GAD and GHD on neurons(1)Establishment of neuron cell injury model and determination of cell survival rate Methods: PC12 cells were induced by glutamate to establish neuron injury model.MTT method was used to detect the cell survival rate after GAD and GHD intervention,and the effect of GAD and GHD on the proliferation of neurons was analyzed.(2)Study on the protective mechanism of GAD and GHD on PC12 cells Enzyme linked immunosorbent assay(ELISA)was used to detect the release of LDH,the content of SOD and GSH.Flow cytometry was used to detect the level of ROS.RT-RCR was used to detect the effect of gardenia blue on tau,BDNF and cred m RNA transcription.The protective effect and mechanism of gardenia blue on neurons were analyzed.Results:1.Study on the complexation of genipin with seven amino acidsThe ratio of genipin to different amino acids was 1:1,1:2,1:3(mol:mol)The higher the ratio of amino acid to genipin,the faster the complexation speed.Considering the residue rate of amino acid,the lowest molar ratio of 1:1 was selected at 80 ℃(mol:mol)It was used in the follow-up experiment.2.The antidepressant effect of gardenia blue on cums rats and its mechanism(1)The results of behavioral studies showed that seven kinds of gardenia blue could increase the consumption of sugar water in cums rats,while only GAD and GHD could prolong the total distance and central movement distance of open field activities,shorten the immobility time of forced swimming and tail suspension,and showed better antidepressant effect,while the other five kinds of gardenia blue could only improve part of depression like behavior,So GAD group and GHD group were selected to further analyze the changes of structure and function of rat flora.(2)The results of hippocampal tissue observation showed that the hippocampal neurons of depression model rats were damaged.Among the seven kinds of gardenia blue,GAD had the most obvious repair effect on the damaged hippocampal neurons,followed by GHD,and the other five kinds of gardenia blue also had weak repair effect;The contents of 5-HT and Da decreased in depression model rats.Different gardenia blue complexes have different degrees of regulation on the contents of 5-HT and DA in brain tissue and serum.(3)The structure of intestinal flora of depression model rats was unbalanced,and the carbon source metabolism was significantly different from that of the control group(P < 0.01,P < 0.05);GAD can inhibit the proliferation of Lactobacillus and Bacteroides,GHD can inhibit the proliferation of Enterobacter and Clostridium perfringens,and promote the proliferation of Bacteroides and lactic acid bacteria,so that the intestinal flora is in a relatively balanced state.3.The protective effect and mechanism of GAD and GHD on PC12 cell injury model induced by Glu The IC50 was determined by MTT method.GAD and GHD can increase the survival rate of injured cells to a certain extent.Both GAD and GHD can down regulate the levels of LDH and ROS in PC12 cell injury model induced by GLU.GAD can reduce the level of tau,increase the level of BDNF and increase the level of CREB;GHD decreased tau level.These results suggest that GAD and GHD have certain effects on neural plasticity,and thus exert their antidepressant effectsConclusion: by considering the antidepressant effects of seven kinds of gardenia blue,the results show that GAD and GHD are the best,which can improve depression like behavior,regulate DA and 5-HT neurotransmitters,repair damaged hippocampal tissue,and regulate intestinal flora of depression model rats;GAD and GHD play a neuroprotective role by inhibiting the production of LDH and ROS,promoting the production of SOD and GSH,and affecting the expression of tau,BDNF and CREB m RNA.