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Objective:To explore the role and mechanism of kidney brain protein (KIBRA) down-regulation in cognitive dysfunction caused by chronic cerebral hypoperfusion.Methods:Ninety male SPF grade Sprague Dawley (SD) rats were divided into four groups according to random number table: sham operation group ( n=15), chronic hypoperfusion group (2VO group, n=25), chronic hypoperfusion stereotaxic injection of AAV-KIBRA group (2VO+ AAV-KIBRA group, n=25), chronic hypoperfusion stereotaxic injection of AAV-Vector group (2VO+ AAV-vector group, n=25). Chronic cerebral hypoperfusion model was established by bilateral ligation of common carotid artery, and stereotactic injection of 2 μL AAV-KIBRA or AAV-vector was performed for 30 days.Morris water maze, in vitro electrophysiology, p21-activated kinase 3(PAK3) activity detection, Western blot, immunoprecipitation and Golgi staining were used to detect spatial learning and memory ability, long-term potentiation(LTP), KIBRA level expression, PAK3 activity changes and the distribution of dendritic spines.SPSS 16.0 statistical software was used for statistical data.One-way ANOVA was used to compare the differences between groups.LSD test was used to compare the significance of data differences between the two groups.Welch test was used for uneven variance. Results:After 1 month of chronic cerebral hypoperfusion, the level of KIBRA in the hippocampus of rats was detected by homogenate and Western blot, and it was found that the level of KIBRA in 2VO group was lower than that of sham group(73.49±4.12)% ( P<0.01). AAV-KIBRA injection in hippocampal CA1 region significantly up-regulated the level of KIBRA to (91.91±7.01)% over 2VO group ( P<0.01). Morris water maze test showed that the latency of the 2VO group(3rd-7th day trail data: (48.18±2.82)s, (43.45±2.27)s, (32.27±2.22)s, (26.55±2.37)s, (17.18±2.67)s) were significantly longer than those of the sham group((41.67±2.74)s, (32.58±2.57)s, (22.50±2.94)s, (16.91±2.39)s, (8.75±1.52)s) (all P<0.05), and the latencies of the 2VO+ AAV-KIBRA group 3rd-7th day trail data: (43.83±2.95)s, (35.25±2.15)s, (26.58±2.03)s, (19.92±2.17)s, (17.75±1.35)s) was significantly shorter than that of the 2VO group ((all P<0.01). The Morris water maze test with the platform removed showed that the latency of rats in the 2VO group to reach the platform region was significantly longer than that of the sham group, while the latency of rats in the 2VO+ AAV-KIBRA group to reach the platform region was significantly shorter than that in the 2VO group ( P<0.01). At the same time, the retention time and the crossing times in the platform region of 2VO group were less than those of the sham group ( P<0.01), but the retention time and the crossing times in the platform region of 2VO+ AAV-KIBRA group were significantly higher than those in the 2VO group ( P<0.01). The electrophysiological records of the brain slices showed that the relative excitatory postsynaptic field potential of 2VO group (1.43±7.43) was significantly lower than that of sham group (2.21±6.54) after high frequency stimulation, while the relative excitatory postsynaptic field potential of 2VO+ AAV-KIBRA group (1.90±8.15) was higher than that of 2VO group ( P<0.01). Immunoprecipitation in rat hippocampus revealed that PAK3 could be detected by Western blot assay when KIBRA was precipitated.The results showed that the relative enzyme activity of PAK3 in 2VO hippocampal tissue (0.64±0.04) was significantly lower than that in sham group (1.02±0.07), while the relative enzyme activity of PAK3 in 2VO+ AAV-KIBRA group (0.86±0.03) was significantly higher than that in 2VO group.Golgi staining showed that the density of dendritic spines in 2VO hippocampal neurons((6.85±0.43)/10 μm) was significantly lower than that in sham group((11.83±0.58)/10 μm), while the density of dendritic spines in 2VO+ AAV-KIBRA group((10.22±0.39)/10 μm) was significantly higher than that in 2VO group. Conclusion:The down-regulated of KIBRA after chronic cerebral hypoperfusion plays a key role in cognitive dysfunction and is also involved in the decrease of synaptic functional plasticity.The downregulation of KIBRA is involved in the structural plasticity of dendrites through the regulation of PAK3 activity.Therefore, KIBRA may be an important target for the prevention and treatment of cognitive function of chronic cerebral hypoperfusion.
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Objective To explore whether pravastatin (Pra) inhibits mammalian target of rapamycin (mTOR) signal pathway by regulating Ras homolog enriched in brain (Rheb) protein through the comparison of gene and protein expression changes of Rheb in liver and placenta in preeclampsia (PE)-like mouse model treated with Pra. Methods C57BL/6J pregnant mice were randomly divided into two groups. The PE group was established by injecting N-nitro-L-arginine methyl ester (L-NAME) daily at gestational 7-18 days, saline was injected as contol group (Con);then giving mice Pra (PE+Pra, Con+Pra group, n=8) or normal saline (PE+N, Con+N group, n=8) every day from the 8th gestational day of pregnancy. The maternal liver and placenta tissues were collected on the 18th day of pregnancy. Western blot, real-time quantitative PCR and immunohistochemistry were used to compare the levels of Rheb protein and mRNA expression in the liver and placenta. Results (1)The results of western blot:there were no significant differences in Rheb protein expression between PE+N group (liver:0.706±0.123;placenta:0.866±0.128) and Con+N group (liver:0.732 ± 0.123; placenta: 0.909 ± 0.097), and the differences between PE+Pra group (liver: 0.669 ± 0.134;placenta:0.940 ± 0.221) and PE+N group were not significant either in liver or in placenta (all P>0.05). (2) The results of real-time quantitative PCR:when PE+N group (liver:1.026 ± 0.480;placenta:1.102 ± 0.361) compared with Con+N group (liver:1.058±0.389;placenta:1.067±0.400), PE+Pra group (liver:0.735±0.356;placenta:0.822±0.304) compared with PE+N group, there were no significant differences either in liver or in placenta (all P>0.05). (3) The results of immunohistochemistry: Rheb protein expression did not change significantly in maternal liver and placenta, there were no significant differences in protein expression levels between PE+N group and Con+N group, and between PE+Pra group and PE+N group (all P>0.05). Conclusion The inhibition of Pra on mTOR signaling pathway in some PE-like model may be independent of the expression of Rheb gene and protein.
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Objective:To detect the repair effect of brain protein hydrolyzate(BPH)on the oxidative damage induced by H2O2in the PC12 cells,and to optimize the prescription of BPH stomach floating tablets using the star design effect surface method.Methods:The PC12 cells in the logarithmic phase were divided into normal control group,model group(300 μmol·L-1H2O2),BPH group,artificial gastric juice-treated BPH(GBPH)group, artificial intestinal juice-treated BPH(IBPH)group,artificial gastric juice-treated BPH tablets(GBPH-T)group and artificial gastric juice-treated BPH floating tablets(GBPH-FT)group(The latter five groups were added with 20,40,60,80 and 100 mg·L-1BPH treated with different conditions);at the same time blank control group was set up.The PC12 cells in normal control group didn't receive any treatment,the blank control group was added with medium only,and the PC12 cells in model group were treated with H2O2(300 μmol·L-1)for 3 h. The cell viability was detected by MTT assay.Using Design-expert 8.0.6 Trial software,the dosages of HPMC-K4M,octadecanol and acrylic resin Ⅱ were used as the investigation factors,and the 8 h cumulative release in vitro was used as the evaluation index to optimize the prescription.Results:Compared with normal control group, the activity of PCl2cells in 60 mg·L-1BPH group was increased(P<0.05).Compared with model group,the vitalities of PC12 cells in BPH group,IBPH group,GBPH group and GBPH-FT group were increased significantly (P<0.05 or P<0.01).Compared with BPH group,the cell viability in IBPH group was decreased significantly (P<0.05),and there was no significant difference in GBPH group(P>0.05).Compared with GBPH-T group, the cell viability in GBPH-FT group was significantly increased(P<0.05).The optimal prescription was BPH 20 mg,lactose 10 mg,magnesium stearate 0.4 mg,microcrystalline cellulose 20 mg,HPMC-K4M 27 mg, octadecanol 63 mg,acrylic resinⅡ13 mg;all floating tablets drift in 5 s and sustained floating > 8 h;the difference of the measured value of 8 h cumulative release and the predicted value was not statistically significant (P>0.05).Conclusion:BPH has a significant repair effect on the H2O2-induced oxidative damage in the PC12 cells.Star design-effect surface method has good predictability and reproducibility;it is reasonable and feasible, and can be used to optimize the prescription of BPH stomach floating tablets.
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Objective: To detect the repair effect of brain protein hydrolyzate (BPH) on the oxidative damage induced by H2O2 in the PC12 cells, and to optimize the prescription of BPH stomach floating tablets using the star design effect surface method Methods: The PC12 cells in the logarithmic phase were divided into normal control group, model group 300 μmol · L-1 H2O2), BPH group, artificial gastric juice-treated BPH (GBPH) group, artificial intestinal juice-treated BPH (IBPH) group, artificial gastric juice-treated BPH tablets (GBPH-T) group and artificial gastric juice-treated BPH floating tablets (GBPH-FT) group (The latter five groups were added with 20, 40, 60, 80 and 100 mg · L-1 BPH treated with different conditions); at the same time blank control group was set up. The PC12 cells in normal control group didn't receive any treatment, the blank control group was added with medium only, and the PC12 cells in model group were treated with H2O2 300 μmol · L-1) for 3 h. The cell viability was detected by MTT assay. Using Design-expert 8.0.6 Trial software, the dosages of HPMC-K4M, octadecanol and acrylic resin E were used as the investigation factors, and the 8 h cumulative release in vitro was used as the evaluation index to optimize the prescription. Results: Compared with normal control group, the activity of PC12 cells in 60 mg · L-1 BPH group was increased (P0.05). Compared with GBPH-T group, the cell viability in GBPH-FT group was significantly increased (P 8 h; the difference of the measured value of 8 h cumulative release and the predicted value was not statistically significant (P>0.05). Conclusion: BPH has a significant repair effect on the H2O2-induced oxidative damage in the PC12 cells. Star design-effect surface method has good predictability and reproducibility; it is reasonable and feasible, and can be used to optimize the prescription of BPH stomach floating tablets.
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Objective:To investigate the efficacy and safety of brain protein hydrolysate injection in the treatment of elderly patients with parkinson.Methods:120 patients with parkinson were selected and randomly divided into two groups.The control group (57 cases) was treated by routine treatment,while the observation group (63 cases) was given brain protein hydrolysate injection on the basis of routine treatment.The UPDRS scores,SF-36 scores and adverse reactions during treatment were observed and recorded.Results:Before treatment,the UPDRS and SF-36 scores of both groups had no significant difference (P＞0.05).After treatment,the UPDRS scores of both groups were significantly decreased,and the UPDRS score of observation group was lower than that of the control group(P＜0.05).The social function,emotional function,mental health score of observation group were higher than those of the control group(P＜0.05).During treatment,1 cases of dizziness,1 cases of nausea were found in the control group.The adverse reaction rate was 3.5％.2 cases of insomnia,1 cases of fatigue,1 cases of dizziness and 1 cases of nausea were found in the observation group.The adverse reaction rate was 7.9％.There was no statistical difference in the incidence of adverse reactions between two groups (P＞0.05).Conclusion:Brain protein hydrolysate injection had significant effect on the Parkinson.It could improve the patients' thinking ability,mental health and quality of life with high safety.
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Objective To explore the therapeutic effect of Xiefu Zhuyu decoction joint brain protein hydrolysate injection in the treatment of patients with traumatic brain injury and its effect on cerebrospinal fluid of endothelin-1 ( ET-1 ) .Methods 102 cases of patients with craniocerebral injury were divided into control group (n=51) and experimental group (n=51),the control group were treated with brain protein hydrolyzate injection, the experimental group based on the control group were treated with Xuefu Zhuyu decoction, the curative effect, ET-1, inflammatory factor, protein, blood rheology, S-100β, neuron specific enolase (NSE), National Institutes of Health Stroke Scale (NIHSS) and complications of two groups were compared.Results The total effective rate of experimental group was higher than the control group (94.11% vs.78.43%), the difference was statistically significant (P<0.05).The ET-1 of experimental group was lower than the control group[(32.45 ±4.05) ng/L vs.(28.11 ±3.50) ng/L] (P<0.05).The inflammatory factor, protein, blood rheology, NSE, S-100βof experimental group were better than those of control group (P<0.05). The NIHSS of experimental group was lower than that of the control group ( P <0.05 ) .The complication rate had no difference between two groups. Conclusion The clinical curative effect of Xiefu Zhuyu decoction joint brain protein hydrolysate injection in the treatment of traumatic brain injury , can reduce cerebrospinal fluid levels of ET-1 and can improve inflammatory factors, proteins and hemorheology.
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ObjectiveTo explore effective treatment for moderate and severe neonatal hypoxic-ischemic encephalopathy(HIE).Methods46 cases of moderate to severe HIE were randomly divided into two groups.Control group of 21 cases only accepted the HIE conventional treatment,treatment group,25 cases accepted the HIE conventional treatment and were also given brain protein hydrolyzate combined naloxone treatment,the efficacy of various clinical indicators of the two groups were compared.ResultsThe remarkable effective rate and the total effective rate of the treatment group were significantly higher ( all P ＜ 0.01 ).Consciousness recovery time,recovery time of the original ability,convulsions,muscle tension,recovery time,sucking ability,recovery time,hospital days of treatment group were significantly lower than the control group( P ＜ 0.05 or P ＜ 0.01 ).ConclusionThe effect of naloxone therapy for moderate to severe HIE is significant,and the therapy has high clinical value.
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Objective:To investigate the effects of Yizhi Oral Solution on protein and RNA synthesis of brain tissue of mice. Methods: The radioactive intensities of 3H leucine in brain protein and 3H uridine in RNA of brain tissue of young mice were observed by the methods of incorporation in vivo and group control observation.Results: The radioaetive intensity of 3H leucine of Yizhi Oral Solution group was higher than that of cycloheximide pathological model group ( P