Objective To investigate and validate the expression profiles of Ppp3cb and Ppm1g through differential proteomic analysis of hippocampal tissue in NHE1 gene knockout mice with proteomic analysis.Method ① Six 2-week-old NHE1 knockout mice were selected as the model group,and 6 wild-type mice of the same age served as the control group,and their genotypes were detected by agar-gel electrophoresis.Open field test and forced swimming test were used to evaluate the behaviors of mice in the model group and control group,and epileptic seizure was graded according to Racine scoring.② Tandem mass spectrometry was employed to screen the differential proteins in the hippocampus tissues from the model group and the control group.Then the obtained differential proteins were annotated and enriched in the Gene Ontology(GO)database.Search tool for the retrieval of interesting genes(STRING)database was used to analyze protein-protein interaction(PPI)among different proteins.③ The transcriptional and translational levels of Ppp3cb and Ppm1g were detected by qPCR and Western blotting,respectively,and their expression levels in the tissues were observed with immunohistochemistry.Results ① NHE1 was not expressed in the model group.The mice of the model group had shorter total movement distance(P=0.007 3)and less crossing cells(P＜0.000 1)in open field test,and longer period of immobility in forced swimming test(P＜0.000 1)when compared with those from the control group.② When fold change ≥1.2 times and P＜0.05 were set as the significant threshold for differential expression,845 differentially expressed protein sites were detected in the hippocampus,among which 9 proteins(including Ppm1g)were up-regulated and 7 ones(including Ppp3cb)were down-regulated.Gene Ontology(GO)functional analysis showed that after NHE1 knockout,the most significant differences were observed in the concentration of molecular function(MF)related to protein serine/threonine phosphatase activity,concentration of cellular component(CC)related to the plasma membrane,and concentration of biological process(BP)related to negative regulation of biological processes and immune system processes.STRING analysis indicated that the differential proteins Ppp3cb and Slc9a1 directly acted,Ppm1g indirectly acted through Ppp3cb and Slc9a1,and Ppp3cb and Ppm1g interacted.③The transcriptional and translational levels of Ppp3cb were decreased,and its expression level was reduced in the tissues,while those of Ppm1g were increased,and its expression was elevated in the tissues(P＜0.05).Conclusion In the hippocampus of NHE1 gene knockout mice,the expression of differential protein Ppp3cb is down-regulated and that of Ppm1g is up-regulated,which provide a basis for further study on their involvement in the pathogenesis of epilepsy.

Objective To observe the effect of TCM compound of resolving stagnation for tranquilization ( RST) on neurotransmitters and brain derived neurotrophic factors ( BNDF ) in depression rat model induced by chronic unpredictable stress ( CUS) , and to reveal the mechanism of RST on the treatment of depression.Methods Altogether 60 male SD rats were randomly divided into normal group, model group, high-, mid-and low-dose RST groups at dosage of 8.2, 16.3, 32.7 g/kg, and fluoxetine group with dose of 10 mg/kg.The rats of four therapy groups were administered intragastrically drugs with undergoing CUS for consecutive thirty-five days.The behavioral changes of rats were observed by open field test and novelty-suppressed feeding-test.The content of dopamine ( DA) , 5-hydroxeytryptamine(5-HT) , 5-hydroxyindole acetic acid (5-HIAA) and norepi-nephrine( NE) in hippocampus and cortex were determined by HPLC.And the expression of BDNF in hippocampus was measured by Western blot.Results Compared with the normal group, the rats ’ weight increased slowlier and the spontaneous activity decreased in model group; the contents of 5-HT and NE in cortex and the content of 5-HT, 5-HIAA and NE in hippocampus significantly reduced ( P<0.05 ).Compared with the model group, the spontaneous activity significantly increased in all therapy groups;the contents of 5-HT and NE in cortex dramatically increased in fluoxetine group and mid-, high-dose group ( P<0.05 );the contents of 5-HT and 5-HIAA in hippocampus significantly increased in all three RST groups ( P<0.05 ); the content of NE in hippocampus significantly increased in low-and high-dose group ( P<0.05 ).The expression of BDNF in hippocampus significantly increased in fluoxetine group and high-dose RST group ( P<0.05).Conclusion The anti-depression mechanism of TCM compound formulated by principle of resolving stagnation for tranquilization may be related to the effects of increasing neurotransmitters and up-regulation of BDNF.