Résumé
Objective To investigate the effect of maternal separation (MS) on cognitive function in adult male rats through the expression of neuronal nitric oxide synthase (nNOS) in hippocampus, and to reveal the roles of early life stress (ELS) on neural development in rats. Methods Healthy SD pregnant rats (n=12) were randomly divided into maternal separation group (MS group) and control group (NMS group) (n=6 for each group). The newborn rats in the MS group were separated from the mother rats for 3 h every day from postnatal day 3 to 22 whereas no intervention was taken in the NMS group. At the age of 10 weeks, Morris water maze was used to test the learning and memory abilities of two groups of offspring male rats. Neuron immunofluorescence staining was used to examine the number and distribution of neurons in dentate gyrus (DG) of two groups of offspring male rats. Western Blot method was used to detect nNOS, eNOS, Bax/BCL2, Caspase-3 and P53 levels in the hippocampus of the two groups. Ki67/DCX immunofluorescence staining were used to examine the proliferation and differentiation of neurons in the DG area of the hippocampus. TUNEL staining was used to detect the neuronal degeneration and death in the DG area of the hippocampus. Results Behavioral tests showed that the escape latency of male rats in MS group was prolonged, the target quadrant residence time and the number of platform crossing decreased (P<0.05) compared with NMS group. Compared with NMS group, the number of normal and degenerated neurons in hippocampal DG area of MS group had no significant change (P＞0.05). However, the expression of nNOS and eNOS in hippocampus was decreased (P<0.05) and the expression of Bax/BCL2 was increased (P<0.05), but the expression of caspase-3 and P53 remained unchanged (P＞0.05). In addition, Neuronal proliferation and differentiation were decreased and apoptosis was increased in MS group (P<0.05). Conclusion Repeated MS reduces the expression levels of nNOS in the hippocampus, affects the neuronal function in the DG area, and has a long-term influence on the neurodevelopment, which results in cognitive deficits related to learning and memory abilities in adult rats.
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Objective To investigate the expression of neural-nitric oxide synthase (nNOS) in renal clear cell carcinoma and its clinical significance.Methods The expression of nNOS mRNA in 533 samples of TCGA database was analyzed with Student t test,and statistical analysis was performed to assess the relationship between nNOS expression and clinical prognosis with Kapla-Meier test.Western blot analysis of nNOS protein expression in 10 cases of clear cell renal cell carcinoma(ccRCC) from department of urology of Wuhan union hospital with student t test.Results The mRNA levels of nNOS in 72 cases of ccRCC in tumor tissues and adjacent tissues and were 2.99 ± 0.28 and-1.57 ± 0.17,it is significantly lower than those in adjacent tissues (P ＜ 0.01).The mRNA levels of nNOS in 533 cases of ccRCC,in tumor tissues and adjacent tissues and were 2.99 ± 0.28 and-1.76 ± 0.05,it is significantly lower than those in adjacent tissues (P ＜ 0.01).A total of 533 sample studies showed a low correlation between nNOS expression and clinical T stage,T1-1.59 ±0.08,T2-1.96 ±0.13,T3-1.90 ±0.09,T4-2.38 ±0.28 (P =0.0029) and -1.63 ±0.06 and-2.16 ± 0.13 between non-metastasis and no-metastasis (P =0.0009),and-1.57 ± 0.08 and-2.03 ± 0.11 between non-recurrence and recurrence (P =0.008).Survival analysis showed that the overall survival time were (40.3 ± 5.6) months and (48.3 ± 5.7) months in lower and higher nNOS expression,and disease free survival time were (37.1 ± 2.1) months and (40.3 ± 5.6) months in lower and higher nNOS expression,both with shorter time in low expression of nNOS (P ＜ 0.01).nNOS proteins were 1.02 ± 0.16 and 0.61 ± 0.1 1 in tumor tissues and adjacent tissues with significantly lower expression(P＜0.05).Conclusions The mRNA and protein of nNOS are lower in ccRCC with a poor prognosis of ccRCC.
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Objective To investigate the effect of propofol on nNOS expression after focal cerebral ischemia-reperfusion in rats and the possible mechanism of protective effect of propofol on brain. Method Seventy-eight male Wistar rats, weighting 250 ~ 300 g, were randomly divided into 3 groups:(1)Sham operation group (S group, n=6) was performed with scham operation; (2) Ischemia-reperfusion group (group I-R, n=36) was subjected to 2-hour right middle cerebral artery occlusion and then reperfusion was followed, saline (1 mg/kg) was injected into the right lateral cerebral ventricle using microsyringe before reperfusion;(3) Propefol group (group P, n=36) was injected with propofol (1mg/kg) into the right lateral cerebral ventricle using microsyringe right after ischemia. Group I-R and group P were divided into 3 subgroups according to the reperfusion time: 1 h, 3 h and 6 h. The neurological function of all rats were tested before reperfusion. The cerebral infarction area of the whole brain was calculated with TIC staining (n=6). The pathological change of brain was observed from HE staining (n=6) and the nNOS protein expression was obtained by immuno- histochemical method (n=6). Results Compared with I-R group, the neurological function was better in group P(P
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Objective To investigate the effect of high power microwave(HPM) irradiation on neuropeptide Y (NPY) and neural nitric oxide synthase (nNOS) expression in the cerebral cortex and hippoeampus of Wistar rats. Methods A total of 110 Wistar rats were used for this study.Three groups of 30 Wistar rats were exposed to HPM irradiation at intensities of 3,10,30 and 100 mW/cm~2,respectively.Twenty rats served as controls and were ex- posed to sham HPM irradiation.At 6 h,and at 1,3,7,14 and 28 d after irradiation,five rats from each group were sacrificed,and their cerebral cortices and hippocampi were harvested.HE staining was used to highlight any change in the structure of the cerebral cortex or hippocampus.Immunohistochemistry techniques and image analysis were used to study the changes in NPY and nNOS expression.Results 10 to 100 mW/cm~2 HPM irradiation caused pyc- nosis and deep staining of some neurons in the cerebral cortex and hippocampus.The increase in nNOS expression and decrease in NPY expression observed were significant at 3 days after irradiation.Conclusion HPM irradiation can induce injury in neurons of the cerebral cortex and hippoeampus,and abnormal NPY and nNOS expression.