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 investigate the efficacy and mechanism of Jasminoside in dextran sodium sulfate (DSS) -induced ulcerative colitis (UC) mice.Methods:Eighteen C57BL/6 mice were randomly divided into control group, UC group and UC+Jasminoside group, with six mice in each group. The mice in UC group and UC+Jasminoside group freely drank 3%DSS solution for 7 days to induce UC mouse model. The mice in control group freely drank the water without DSS. During the modeling period, the mice in the UC+Jasminoside group were given Jasminoside at a dose of 20 mg/ (kg·d) by gavage for 7 consecutive days, meanwhile the mice in control group and UC group were given equal volumes of distilled water by gavage. The change of body weight and disease activity index (DAI) of the mice were observed daily, and the length of the colon was measured 7 days after the intervention. HE staining was used to observe the inflammatory changes of the colon, and Western blot was used to measure the expression of anti-apoptotic proteins (Bcl-2 and Bcl-xl), caveolin-1 (CAV-1), inducible nitric oxide synthase (iNOS), tight junction proteins (Occludin and Claudin-1). The differences in the above indicators among the mice in three groups were analyzed statistically.Results:All the mice survived. Compared with the control group, the body weight was lower, DAI score was higher, colon was shorter, whose differences were all statistically significant (all P<0.05), and inflammatory infiltration of colon tissue was more severe in mice of UC group. Compared with the UC group, body weight was higher, DAI score was lower, colon was longer, whose differences were all statistically significant (all P<0.05), and inflammatory infiltration of colon tissue was alleviated in mice of UC+Jasminoside group. Western blot results showed that compared with control group, the expression of Bcl-2, Bcl-xl, CAV-1, Occludin and Claudin-1 was significantly lower and the expression of iNOS was significantly higher in the colon tissues of mice in UC group, and the differences were statistically significant (all P<0.05). The expression of Bcl-2, Bcl-xl, CAV-1, Occludin and Claudin-1 in the colon tissues of mice in UC+Jasminoside group was significantly higher than those in UC group, while the expression of iNOS was lower, and the differences were statistically significant (all P<0.05) . Conclusion:Jasminoside can alleviate the symptoms of UC model mice, and its mechanism of action may be related to upregulating the expression of anti-apoptotic proteins and protecting the intestinal mucosal barrier through CAV-1/iNOS pathway.

Objective:To investigate the efficacy and mechanism of Jasminoside in dextran sodium sulfate (DSS) -induced ulcerative colitis (UC) mice.Methods:Eighteen C57BL/6 mice were randomly divided into control group, UC group and UC+Jasminoside group, with six mice in each group. The mice in UC group and UC+Jasminoside group freely drank 3%DSS solution for 7 days to induce UC mouse model. The mice in control group freely drank the water without DSS. During the modeling period, the mice in the UC+Jasminoside group were given Jasminoside at a dose of 20 mg/ (kg·d) by gavage for 7 consecutive days, meanwhile the mice in control group and UC group were given equal volumes of distilled water by gavage. The change of body weight and disease activity index (DAI) of the mice were observed daily, and the length of the colon was measured 7 days after the intervention. HE staining was used to observe the inflammatory changes of the colon, and Western blot was used to measure the expression of anti-apoptotic proteins (Bcl-2 and Bcl-xl), caveolin-1 (CAV-1), inducible nitric oxide synthase (iNOS), tight junction proteins (Occludin and Claudin-1). The differences in the above indicators among the mice in three groups were analyzed statistically.Results:All the mice survived. Compared with the control group, the body weight was lower, DAI score was higher, colon was shorter, whose differences were all statistically significant (all P<0.05), and inflammatory infiltration of colon tissue was more severe in mice of UC group. Compared with the UC group, body weight was higher, DAI score was lower, colon was longer, whose differences were all statistically significant (all P<0.05), and inflammatory infiltration of colon tissue was alleviated in mice of UC+Jasminoside group. Western blot results showed that compared with control group, the expression of Bcl-2, Bcl-xl, CAV-1, Occludin and Claudin-1 was significantly lower and the expression of iNOS was significantly higher in the colon tissues of mice in UC group, and the differences were statistically significant (all P<0.05). The expression of Bcl-2, Bcl-xl, CAV-1, Occludin and Claudin-1 in the colon tissues of mice in UC+Jasminoside group was significantly higher than those in UC group, while the expression of iNOS was lower, and the differences were statistically significant (all P<0.05) . Conclusion:Jasminoside can alleviate the symptoms of UC model mice, and its mechanism of action may be related to upregulating the expression of anti-apoptotic proteins and protecting the intestinal mucosal barrier through CAV-1/iNOS pathway.

Aim To study the therapeutic effect of CpG-ODN, an agonist of Toll-like receptor 9 (TLR9), on hypoxic/ischemic encephapathy in neonatal rats and investigate the mechanisms.Methods Fifty healthy 7-day-old neonatal Wistar rats (in either gender, weighing 12~17g) were randomly divided into sham operation group, HIBD group, and CpG-ODN low group(0.35 mL·kg-1), CpG-ODN middle group(1.40 mL·kg-1), CpG-ODN high group(5.60 mL·kg-1).The neurological function was scored after 48h operation;ten rats of each group was executed respectively and brains tissue was taken;HE staining was used to observe the brain pathological changes.Western blot assay was used to detect the expressions of TLR9 and phosphor-p38 mitogen-activated protein kinases(p-p38 MAPK), and enzyme linked immunosorbent assay (ELISA) method was adopted to detect TNF-α expression.Results The CpG-ODN low, middle group were improved in impairment significantly compared with the HIBD group, and the brain pathological change was lessened, while the CpG-ODN high group was impaired significantly compared with the HIBD group (P<0.05), and brain pathological change was sharpened.Western blot showed the up-regulation in TLR9 and p-p38 MAPK and a significant increase of the expression of TNF-α in the brain tissue in CpG-ODN group with statistical difference in HIBD group and sham operation group(P<0.05).Conclusions The neuro-behavioral score and nervous system function can be improved and the hypoxic/ischemic brain damage can be reduced in neonatal rats in the CpG-ODN low, middle group.The protective mechanisms may be suitably via activating p38 MAPK signaling pathway to promote p38 MAPK phosphory1ation and up-regulation of the expression of TNF-α in the brain tissue of rats.