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Objective To explore the expression changes and possible molecular mechanisms of Neuritin and HSP60 during the repair of liver injury,and to provide an experimental basis for the study of the repair of liver injury. Methods Forty-eight Sprague-Dawley(SD)rats were randomly divided into control group without any treatment(n=6)and experimental group(n=42)underwent 70% hepatectomy to induce acute liver injury,and 6 h,12 h,24 h,48 h,3 d,7 d and 14 d after the operation,the left lobe resection of the residual liver was per-formed. Immunoblotting technique (Western blot) was used to detect the expression difference of Neuritin and HSP60 in liver tissue of the corresponding time points. Hematoxylin eosin staining(hematoxylin-eosin staining, HE)was used to observe the expression changes at each time point in liver pathology,and tail vein blood to detect ATL and AST changes. Results (1)Compared with those in the control group,the serum ALT and AST levels in the experimental group were increased at 6 h,12 h,24 h and 48 h after the operation,and reached the peak at 48 h postoperatively but those began to decrease 3 d postoperatively and was almost normal 7 d postoperatively. The difference was statistically significant(P < 0.05). Pathological findings:compared with that in the control group, the hepatic lobule structure in the experimental group was disorderly. The obvious balloon like change reached the peak 48 h postoperatively.(2)In the whole process of repair of liver injury,the expression of Neuritin and HSP60 showed differences in the opposite. There was a significantly negative correlation between Neuritin expression and the repair or the aggravation of the injury(P<0.001)and the lowest expression was observed 48 h postoperatively. There was a significantly positive correlation between HSP60 expression and the repair or the aggravation of the injury(P < 0.001)and the highest expression was observed 48 h postoperatively. Conclusions (1)With simple 70% left hepatic lobectomy,the repair of liver injury model of SD rats is successfully established and the heaviest injury is observed 48h postoperatively. With a high success rate(100%,42/42),simple and practical,the method provides a reliable and convenient animal model for the study of liver regeneration,liver injury and liver transplan-tation.(2)The expression of Neuritin decreases gradually after liver injury and reaches the lowest 48 h postopera-tively,while the expression of HSP60 increases gradually and reaches the highest 48 h postoperatively.(3)The change of expression of Neuritin and HSP60 is closely related to the process of liver injury repair(P < 0.001), showing a certain change rule. They may have some biological effects through interactions,and participate in and promote the regeneration and repair of liver injury.
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Objective To establish transgenic mice model with over expression of neuritin in bone mar-row,for the further study on the function of neuritin protein in the treatment of peripheral neuropathy. Methods Two pairs of transgenic mice(loxp-stop-loxp-neuritin and lyz-Cre/+)were fed and propagated,the DNA from the mice tails extracted and the genotype of transgenic mice identified by PCR. The wild type mice with B6 were as-signed as the controls,and the immunofluorescence was used to detect the accuracy of the neuritinloxp/+ _lyz -Cre/+. Results The two trensgenetic homozygous mice had the ability to reproduce,and the hybrid offsprings were neuritinloxp/+_lyz-Cre/+,neuritinloxp/-_lyz-Cre/+,neuritinloxp/+_lyz-Cre/-,neuritinloxp/-_lyz-Cre/-. The re-sults were met with the Mendel′s law. The results of immunofluorescence showed that the expression of neuritin of neuritinloxp/+_lyz-Cre/+ mice in bone marrow was significantly higher than the wild type mice(P < 0.05). Con-clusion The PCR method is of high reliability for identification of sub pus genotype and the female neuritinloxp/+mice mating with the male lyz-Cre/+ ones is an effective way for obtaining the neuritinloxp/+_lyz-Cre/+ mice.
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Objective To research the expression of Neuritin in gastric cancer .and the relationship between the expression of Neuritin with the occurrence and development of gastric cancer .Methods Collected 58 surgical specimens of gastric cancer from 2010 to 2013 in the first affiliated hospital of Shihezi University ,immunohistochemistry was used to examine the expression of Neu-ritin in gastric cancer and normal tissues near the cancer of the stomach .Results Immunohistochemical staining showed that Neu-ritin was moderately or highly expressed in 96 .55% (56/58) of gastric cancer ,and Neuritin was moderately or highly expressed in 94 .83% (55/58) of normal tissues near the cancer .There was no statistically significantly difference between two groups (P>0 .05);Neuritin was highly expressed in 82 .76% (48/58) of gastric cancer ,and Neuritin was highly expressed in 15 .52% (9/58) of normal tissues near the cancer ,differences between the two groups had statistically significantly (P 0 .05) .Conclusion The Neuritin expression level between gastric cancer and normal tissues near the cancer exist differences .There is overexpression of neuritin in gastric cancer .
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Objective To detect time-dependent change of neuritin expression in brain tissues after traumatic brain injury and discuss the effect of neuritin after brain damage occurred. Methods Forty-two rats were divided into normal group, control and experimental group. According to the postoperative time divided into 6 subgroups, including 6 hours group, 12 hours group, 24 hours group, 3 days group, 7 days group and 14 days group. Immunohistochemical and western-blot were used to detected the protein expression levels of neuritin. Results The immunohistochemical staining indicated that the positive expression of neuritin was strong in the cytomembrane and cytoplasms of the neurons, with a higher intensity, 6 hours after the operation. 12 hours after the operation last to the seventh day, the neurons with the strongest positive expression, is significantly higher than control group and normal group, significant decrease on the fourteenth day. The result of western-blot indicated that the level of neuritin protein sharply increased at 6 hours, reached the peak on 24 hours and after lasted to the seventh day, significantly higher than control group and normal group (P < 0.01), significant decrease on the fourteenth day (P < 0.05). Conclusion Up-regulation of neuritin in cerebral contusion tissues may play an important role after traumatic brain injury.
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In the present study, we compared the expression patterns of neuritin mRNA in the dentate gyrus of hippocampal formation following single (1xECS) or repeated elctroconvulsive seizures (8xECS) treatments to the rat. The expression of neuritin mRNA was transiently increased, and returned to basal level within 12 hours after 1xECS. Whereas initial induction of neuritin mRNA was similarly seen, these increased mRNA level was maintained for 24 hours in 8xECS group. In contrast, induction profile of BDNF was similar following 1x and 8xECS. These results suggest that regulation of neuritin expression may be plastically changed by repeated ECS treatment.
Subject(s)
Animals , Rats , Brain-Derived Neurotrophic Factor , Dentate Gyrus , Hippocampus , Neuronal Plasticity , Plastics , RNA, Messenger , SeizuresABSTRACT
Neuritin is a new neurotrophic factor found recently. In order to identify the function of Neuritin clearly, the coding sequence of human neuritin was amplified by PCR from neuritin cDNA , this fragment digested by NocI and NotI was inserted into pET32a by T4 ligase and transformed into E. coli BL21 then the recombinant plasmid named pET32a-neuritin was constructed successfully . Neuritin was expressed distinctly after inducing by EPTG. The product was identified as neuritin by SDS-PAGE and Western blot analysis . The expression production was purified on Ni2+-NTA column.