大鼠,新生

摘要： 目的 研究新生大鼠支气管肺发育不良(bronchopulmonary dysplasia,BPD)发生、发展中核转运蛋白KPNA2定位及表达的动态变化,探究其在早产儿BPD发病机制中的作用.方法 采用85％的吸入氧浓度诱导新生SD大鼠BPD模型(n=50),对照组吸入空气(n=50),两组分别于1、3、7、10、14 d采集肺组织标本,分离、纯化及培养肺泡Ⅱ型上皮细胞.利用免疫组化、免疫荧光、Western blot及实时荧光定量PCR检测KPNA2的分布及表达.结果 免疫组化可见KPNA2主要定位于肺泡上皮细胞的胞浆及胞核内,与对照组相比,BPD组1 d胞核、胞浆均表达增加,3～14 d胞核表达明显减弱,胞浆表达增强.细胞免疫荧光可见KPNA2在对照组1～14 d主要定位于细胞核,BPD组3～14 d主要定位于细胞浆,BPD组KPNA2核表达较对照组减弱,胞浆表达较对照组增强.KPNA2总蛋白及浆蛋白表达趋势基本一致,与对照组比较,BPD组1 d开始升高(P＜0.05),3 d达高峰(P＜0.05),7 d开始逐渐下降(P＜0.01),14 d仍高于对照组(P＜0.05);BPD组KPNA2核蛋白表达3 d开始降低(P＜0.01),持续降低至14 d(P＜0.05).与对照组相比,BPD组KPNA2 mRNA表达1 d开始升高(P＜0.05),3 d达高峰(P＜0.05),7d开始逐渐下降(P＜0.01),至14d仍高于对照组(P＜0.05).结论 暴露于高氧中新生鼠KPNA2核转运功能障碍,可能是影响BPD肺泡上皮细胞DNA损伤应答早期启动的重要机制.

摘要： Objective To investigate the mechanism of white matter damage (WMD) and the neuroprotective effect of Xenon on neonates with WMD.Methods Three-day-old SD rat pups (n =96) were randomly divided into the blank control group (n =24),the WMD control group (n =24),the Xenon intervention group A (n =24) and the Xenon intervention group B (n =24) by random number method according to their birth time.WMD rat models were successfully established by giving intraperitoneal injection of lipopolysaccharide(LPS) 0.05 mg/kg combined with carotid artery ligation and hypoxia for 1 hour in the WMD control group and the Xenon intervention groups.In the control group,only 9 g/L saline (0.05 mg/kg) was injected intraperitoneally,while carotid artery ligation and hypoxia were not administered.Rats in Xenon intervention group A and group B were given inhalation of 500 mL/L Xenon for 3 hours at 0 and 2 hours respectively after establishment of the models.Six rats in each group were randomly selected and decapitated at 0,24,48 and 72 hours after the intervention.The brain white matter on the right was analyzed by using HE staining and myelin basic protein(MBP) immunofluorescence staining,and real-time quantitative polymerase chain reaction was used to detect the expressions level of CLIC4 mRNA.Results (1) Brain tissue pathology:compared with the blank control group,the brain white matter on the right of the WMD control group and the Xenon intervention group A and group B had loose and disordered structure,nuclear pyknosis and cytoplasm loosening.However,the lesions in both Xenon intervention group A and group B were significantly less than those in the WMD control group,and there was no significant difference between the Xenon intervention group A and group B.(2) MBP measurement:the number of MBP-positive cells in the brain white matter on the right of WMD control group was significantly lower than that in the blank control group,while compared with WMD control group,they were significantly higher in Xenon intervention group A and group B.(3) CLIC4 mRNA expression level:compared with blank control group,the expressions levels of CLIC4 mRNA at most time point were higher both in the WMD control group and the Xenon intervention group A and group B (all P ＜ 0.05),except the time point 24 h in the Xenon intervention group A.The expressions of CLIC4 mRNA in group A and group B were significantly decreased compared with those in the WMD control group (all P ＜ 0.05).However,there were no significant differences between Xenon intervention group A and group B (P ＞ 0.05).Conclusions The expressions of CLIC4 mRNA in brain tissues on neonatal rats with WMD significantly increased,indicating that the mitochondrial pathway could be one of the pathological processes of WMD.Early Xenon intervention may reduce neonatal WMD by reducing the expression of CLIC4 mRNA,which plays a neuroprotective role.%目的 探讨早产儿脑白质损伤(WMD)的发病机制及氙气的神经保护作用机制.方法 将3日龄SD新生大鼠96只按照出生时间编号后采用随机数字表法随机分为空白对照组(24只)、WMD对照组(24只)、氙气干预A组(24只)和氙气干预B组(24只).WMD对照组及氙气干预2组予腹腔注射脂多糖(LPS)0.05 mg/kg,并结扎右侧颈总动脉联合缺氧1h处理,建立WMD新生大鼠模型;空白对照组仅腹腔注射9g/L盐水(0.05 mg/kg),不给予颈动脉结扎和缺氧处理.氙气干预A组和B组分别在模型建立后0、2h予500 mI/L氙气吸入处理3h.各组分别于干预处理后0、24、48、72 h采用随机数字表法随机各选取6只大鼠进行断头取脑,给予HE染色,髓鞘碱性蛋白(MBP)免疫荧光染色,实时定量聚合酶链反应检测脑组织中CLIC4 mRNA的表达水平.结果 1.脑组织病理:与空白对照组比较,WMD对照组右侧脑白质结构疏松、紊乱,可见核固缩、胞质疏松等改变;氙气干预2组也均可见上述表现,但较WMD对照组明显减轻,A、B2组间无明显差异.2.MBP测定:WMD对照组右侧脑白质MBP阳性细胞数量较空白对照组明显减少,但氙气干预2组MBP阳性细胞数量较WMD对照组明显增加.3.CLIC4 mRNA表达水平:除了氙气干预A组的24h外,WMD对照组及氙气干预A组和B组右侧脑白质CLIC4 mRNA的表达水平在其余时间点均高于空白对照组,差异均有统计学意义(均P＜0.05);但与WMD对照组相比,氙气干预A组与B组在各时间点CLIC4 mRNA的表达水平均明显降低,差异均有统计学意义(均P＜0.05);氙气干预A组与B组脑组织中CLIC4 mRNA表达水平差异无统计学意义(P＞0.05).结论 WMD新生大鼠脑白质CLIC4 mRNA表达水平明显升高,表明线粒体途径是WMD的病理过程之一;氙气早期干预可通过降低CLIC4mRNA的表达,减轻WMD,从而发挥神经保护作用.

摘要： 目的 观察粒细胞集落刺激因子(G-CSF)对新生大鼠缺氧缺血性脑损伤(HIBD)后炎症反应的影响及哺乳动物雷帕霉素靶蛋白/p70核糖体S6蛋白激酶(mTOR/p70S6K)信号通路在其中发挥的作用.方法 7日龄新生SD大鼠90只按随机数字表法随机分为假手术组、缺氧缺血(HI)模型组、G-CSF组、雷帕霉素组及对照组,改良Rice法建立新生大鼠HIBD模型.HI前1 h,雷帕霉素组腹腔注射雷帕霉素250μg/kg,对照组予等体积乙醇腹腔注射.HI 1 h后,G-CSF组、雷帕霉素组及对照组腹腔注射G-CSF 50μg/kg,HI模型组及假手术组予等体积9 g/L盐水腹腔注射.HI 48 h后,Western blot检测脑组织中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-10及mTOR/p70S6K信号通路蛋白水平,尼氏染色评估海马CA1区及皮质神经元损伤情况,2,3,5-氯化三苯基四氮唑染色检测脑梗死面积.结果 与HI模型组比较,G-CSF组及对照组脑梗死面积减少[(12.87±1.54)％、(11.90±1.31)％比(24.21±3.28)％],尼氏染色阳性神经元计数增多[海马CA1区:(61.00±4.90)个/视野、(61.67±6.40)个/视野比(42.67±4.46)个/视野;皮质:(92.67±6.68)个/视野、(90.17±4.45)个/视野比(70.83±6.97)个/视野],TNF-α及IL-1β水平降低(TNF-α:0.67±0.07、0.55±0.05比0.86±0.05;IL-1β:0.65±0.06、0.52±0.10比0.86±0.06),IL-10、p-mTOR/mTOR及p-p70S6K/p70S6K水平升高(IL-10:0.68±0.04、0.62±0.05比0.34±0.02;p-mTOR/mTOR:0.53±0.02、0.51±0.01比0.26±0.01;p-p70S6K/p70S6K:0.89±0.03、0.90±0.03比0.55±0.02),差异均有统计学意义(均P<0.05).与G-CSF组及对照组比较,雷帕霉素组脑梗死面积增加[(25.70±1.50)％],尼氏染色阳性神经元计数减少[海马CA1区:(40.67±3.50)个/视野;皮质:(68.33±8.17)个/视野],TNF-α及IL-1β水平升高(分别为0.97±0.06、0.98±0.10),IL-10、p-mTOR/mTOR及p-p70S6K/p70S6K水平降低(分别为0.21±0.02、0.30±0.01、0.55±0.01),差异均有统计学意义(均P<0.05).结论 G-CSF可能通过上调mTOR/p70S6K信号通路抑制HIBD后的炎症反应.

摘要： Objective To investigate the effect of docosahexaenoic acid (DHA) on the body weight growth and lipid metabolism of neonatal rats during lactation.Methods The specific pathogen free Sprague-Dawley neonatal rats were randomly assigned into 4 groups (high-dose group,medium-dose group,low-dose group and control group) by random number table method.The rats in 3 experiment groups received intragastric administration with DHA 600 mg/kg,300 mg/kg and 100 mg/kg,respectively,while the control group were given 9 g/L saline,totally for 21 days.Body weight was monitored and compared among groups on postnatal day 1,7,14 and 21.And body weight growth rates at each time point were calculated.The serum concentrations of high density lipoprotein cholesterol (HDL-C),low density lipoprotein cholesterol(LDL-C),triglyceride (TG) and total cholesterol were measured and compared at 6-week and 8-week ages.The pathological and histological changes in the heart,the large vessel and the liver were observed at same time.Results The mean body weight of the neonatal rats were significantly different among 4 groups on postnatal day 7,14 and 21 (F =17.334,4.159,6.485,all P ＜ 0.01).Comparisons were made between every 2 groups,the low-dose group was higher than the control group on postnatal day 7 [(21.60 ±0.89) g vs.(18.57 ± 0.76) g] and day 21 [(58.52 ±6.62) g vs.(53.01 ± 11.75) g];the medium-dose group was lower than the control group on postnatal day 7 [(14.23 ±0.49) g vs.(18.57 ±0.76) g] and lower than the low-dose group on postnatal day 21 [(52.47 ±8.18) g vs.(58.52 ±6.62) g];the high-dose group was lower than the low-dose group on postnatal day 7[(16.13 ± 1.02) g vs.(21.6 ±0.89) g],and it was lower than the control group and the low-dose group on postnatal day 14[(31.69 ± 1.77) g vs.(37.60 ± 1.32) g and (36.24 ±0.84) g],and lower than all the other 3 groups on postnatal day 21 [(45.9 ± 13.17) g vs.(53.01 ± 11.75) g,(58.52 ±6.62) g and (52.47 ±8.18) g];all the differences above were statistically significant (all P ＜ 0.05).During the first and the second week after birth,there were significant differences in the mean body weight growth rate among 4 groups (F =8.369,8.331,all P ＜ 0.01),but there was no significant difference during the third week (F =0.603,P ＞ 0.05).Compared with 2 groups,the mean body weight growth rate of the low-dose group was higher than that of the control group in the first week [(184.96 ± 63.16) ％ vs.(141.02 ± 72.07) ％],but which was lower than that of the control group in the second week [(72.60 ± 35.37) ％ vs.(103.20 ± 40.11) ％];the medium-dose group was lower than the low-dose group at the first week [(116.78 ± 51.59) ％ vs.(184.96 ± 63.16)％],but higher than the low-dose group and lower than the control group at the second week[(139.93 ± 67.4) ％ vs.(72.60 ± 35.37) ％ and (103.20 ± 40.11) ％];the high-dose group was lower than the low-dose group in the first week [(137.33 ± 34.42) ％ vs.(184.96 ± 63.16) ％] and lower than that of the medium-dose group in the second week [(98.22 ± 65.86) ％ vs.(139.93 ± 67.4) ％];all these differences were statistically significant (all P ＜ 0.05).At 6 weeks of age,the mean serum concentrations of total cholesterol,TG and LDL-C were not significandy different (F =1.899,1.450,2.581,all P ＞ 0.05) among 4 groups,but the mean concentration of HDL-C was statistically different (F =7.801,P ＜ 0.01).In detail,the mean concentration of HDL-C in medium-dose group was higher than that of the control group,the low-dose group and the high dose group [(1.66 ± 0.08) mmol/L vs.(0.97 ± 0.16) mmol/L,(1.20 ± 0.09) mmol/L and (0.82 ± 0.09) mmol/L,all P ＜ 0.05],and which in the high-dose group was lower than that in the low-dose group (P ＜ 0.05).At 8-week age,the mean serum concentrations of HDL-C,LDL-C and total cholesterol were not significantly different among 4 groups (F =0.935,0.300,1.299,all P ＞ 0.05),but the mean concentration of TG was significantly different (F =2.875,P ＜ 0.05).The mean concentration of TG in the medium-dose group was lower than that in the control group [(0.98 ± 0.11) mmol/L vs.(1.36 ± 0.09) mmol/L,P ＜ 0.05].There were 5 (15.62％) neonatal rats in the high-dose group which were found to have adipose tissue accumulation around the large vessel walls and the heart and were confirmed by histological examination.The liver cells in these rats were found to have mild fatty changes.No similar changes were found in the other groups.Conclusions Neonatal rats supplemented with DHA during lactation can affect their body weight growth and lipid metabolism.Supplemented with high dose may bring risks,while moderate dose may bring benefits.%目的 探讨新生大鼠在哺乳期补充二十二碳六烯酸(DHA)对其体质量增长、脂肪代谢的影响.方法 无特定病原体级SD新生大鼠采用随机数字表法随机分为DHA高剂量组、中剂量组、低剂量组与对照组,分别在哺乳期予DHA 600 mg/kg、300 mg/kg、100 mg/kg和9 g/L盐水灌胃,共干预21 d.比较各组仔鼠在出生1d、7d、14 d、21 d的体质量,并计算各时间段的体质量增长率,同时比较出生6周、8周龄时的血清高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、三酰甘油(TG)、总胆固醇水平;观察出生6周、8周龄时心、大血管、肝的大体病理改变和组织学改变.结果 出生7d、14d和21 d,4组仔鼠体质量比较,差异均有统计学意义(F=17.334、4.159、6.485,均P＜0.01);两两比较发现,低剂量组体质量在出生7d、21 d[(21.60±0.89)g、(58.52±6.62)g]高于对照组[(18.57±0.76)g、(53.01±11.75)g];中剂量组体质量在出生7 d[(14.23 ±0.49)g]低于对照组,21 d低于低剂量组[(52.47 ±8.18)g比(58.52 ±6.62)g];高剂量组体质量在出生7d[(16.13±1.02)g]低于低质量组,14 d[(31.69±1.77)g]低于对照组[(37.60±1.32)g]和低剂量组[(36.24±0.84)g],21 d[(45.9±13.17)g]均低于其他3组,差异均有统计学意义(均P＜0.05).出生第1周(1 ～7 d)、第2周(8～14 d),4组仔鼠体质量增长率比较,差异均有统计学意义(F=8.369、8.331,均P＜0.01);但在出生第3周(15～21 d),4组仔鼠体质量增长率比较,差异无统计学意义(F =0.603,P＞0.05).两两比较发现,低剂量组出生第1周体质量增长率[(184.96±63.16)％]高于对照组[(141.02±72.07)％],但在第2周却低于对照组[(72.60±35.37)％比(103.20±40.11)％];中剂量组出生第1周体质量增长率[(116.78±51.59)％]低于低剂量组,但在第2周[(139.93±67.4)％]却高于低剂量组和对照组;高剂量组出生第1周的体质量增长率[(137.33±34.42)％]低于低剂量组[(184.96±63.16)％],第2周[(98.22±65.86)％]低于中剂量组,差异均有统计学意义(均P ＜0.05).出生6周,4组仔鼠的血清总胆固醇、TG和LDL-C浓度比较,差异均无统计学意义(F=1.899、1.450、2.581,均P＞0.05);而HDL-C的浓度比较,差异有统计学意义(F =7.801,P＜0.01).两两比较发现,中剂量组血清HDL-C浓度均高于对照组、低剂量组、高剂量组[(1.66±0.08) mmol/L比(0.97±0.16) mmol/L、(1.20±0.09) mmol/L、(0.82±0.09)mmol/L],差异均有统计学意义(均P＜0.05),而高剂量组血清HDL-C浓度尚低于低剂量组,差异有统计学意义(P＜0.05).出生8周,4组仔鼠血清HDL-C、总胆固醇和LDL-C浓度比较,差异均无统计学意义(F=0.935、0.300、1.299,均P＞0.05);而TG浓度比较,差异有统计学意义(F=2.875,P＜0.05).两两比较发现,仅中剂量组的血清TG浓度[(0.98 ±0.11) mmol/L]低于对照组[(1.36 ±0.09) mmol/L],差异有统计学意义(P＜0.05).高剂量组15.62％(5/32只)的仔鼠出现心脏、大血管壁周围脂肪组织堆积,组织学镜检证实为脂肪颗粒,肝脏细胞呈轻度脂肪样变;其余组未发现类似改变.结论 新生仔鼠在哺乳期补充DHA可影响其体质量增长和脂肪代谢,高剂量补充可能带来风险,中剂量补充可能带来益处.

摘要： 目的 研究缺氧诱导因子-1α(HIF-1 α)在新生大鼠缺氧缺血性脑损伤(HIBD)早期mRNA和蛋白水平的表达变化及其作用.方法 1.实验1:36只7日龄SD大鼠按随机数字表法分为假手术组(Sham组,6只)和模型组(HIBD组,30只),模型组根据HIBD后处死大鼠时间的不同又分为5个亚组(6h、12 h、24h、48 h、72 h,每组各6只).实时荧光定量PCR(qPCR)、Western blot分别检测HIF-1α mRNA和蛋白表达水平.2.实验2:将45只7日龄SD大鼠按随机数字表法分为3组:Sham组、HIBD组、2-甲氧基雌二醇组(2ME2组),每组各15只.依据实验1选取HIF-1αmRNA及蛋白表达水平最高的时间点处死取脑,免疫荧光检测HIF-1α蛋白的分布与表达,HE染色观察脑组织病理形态学变化,干湿重法测定脑组织含水量,原位末端标记法(TUNEL)检测细胞凋亡.结果 HIBD早期HIF-1αmRNA和蛋白表达水平均呈先升高后下降趋势,并于HIBD后24h其mRNA表达水平(3.38±o.21)和蛋白表达水平(2.81±0.36)最高.Sham组HIF-1α蛋白主要在细胞质中表达,HIBD组其主要在细胞核中表达.Sham组、HIBD组和2ME2组3组间在HIF-1α染色阳性细胞数、脑组织含水量、细胞凋亡率的比较差异均有统计学意义(均P＜0.05),且2ME2组均显著低于HIBD组(均P＜0.05),脑组织病理形态学改变也较HIBD组轻.结论 HIBD后24 h HIF-1α mRNA和蛋白表达水平最高,抑制HIF-1α的表达,可减轻缺氧缺血所致的脑损伤,推测HIF-1α在新生大鼠HIBD早期可能起损伤作用.%Objective To study the expression of hypoxia-inducible factor-1a(HIF-1α) at mRNA and protein levels in the early stage of hypoxic-ischemic brain damage (HIBD) in neonatal rats and its role.Methods (1) Experiment 1:thirty-six postnatal 7-day SD rats were divided into Sham group (n =6) and model group (HIBD,n =30) according to the random table method,then the rats in the model group were divided into 5 subgroups according to the time of sacrifice after HIBD(6 h,12 h,24 h,48 h,72 h,n =6).The expression levels of HIF-1cα mRNA and protein were detected by quantitative Real-time PCR(qPCR) and Western blot,respectively.(2) Experiment 2:forty-five postnatal 7-day SD rats were randomized into 3 groups:Sham group (n =15),HIBD group (n =15) and 2-methoxyestradiol(2ME2) group(n =15).According to the experiment 1,at the time point of the highest expression levels of HIF-1 α mRNA and protein,rats were killed and the brains were collected.The location and expression of HIF-1 α protein were detected by immunofluorescence,histopathological changes of brain were observed by HE staining,brain water content was measured by dry-wet method,cell apoptosis was detected by nick end labeling(TUNEL) method.Results At the early stage of HIBD,the expression levels of HIF-1 α mRNA and protein increased at first and then decreased,and the mRNA expression level (3.38 ± 0.21) and protein expression level (2.81 ± 0.36) were the highest at 24 h after HIBD.In Sham group,HIF-1 α protein was mainly expressed in the cytoplasm,while in HIBD group it was mainly expressed in the nucleus.The number of HIF-1α staining positive cells,brain water content and apoptosis rate were significantly different among Sham group,HIBD group and 2ME2 group (all P ＜ 0.05),and which were significantly lower in 2ME2 group than those in HIBD group (all P ＜ 0.05),and the pathological changes were also less serious than those in HIBD group.Conclusions The mRNA and protein levels of HIF-1 α are the highest at 24 h after HIBD.Inhibiting the expression of HIF-1 α can ameliorate the brain damage of neonatal rats induced by hypoxia-ischemia.Therefore,it is hypothesized that HIF-1α may cause injury in the early stage of HIBD in neonatal rats.

摘要： Objective To explore the change of endogenous glucocorticoids (GC) secretion after intracranial hemorrhage (ICH) of neonatal rats and the impact of Dexamethasone (DEX).Methods Ten-day-old Sprague-Dawley rat pups of both sexes were randomized into 11 groups:normal control group(CON group),sham operated group (SHAM group),ICH group(each of which was further subgrouped into 12 h group,24 h group and 72 h group according to execution time after the modeled operation),glucocorticoids receptor (GR) agonist intervene group (DEX group) and GR antagonist intervene group(RU486 group).The intracranial autologous blood injection model of ICH was employed.Neurological functional deficits was measured by neurological deficit score (NDS),the levels of cerebral homogenate GC were tested by the emission immunology method,and the pathologic change and the expression of GR in hippocampus CA1 were examined by using Nissl staining and immunofluorescence separately.Results (1) Seventy-two-hour after the modeled operation,NDS of rats in the ICH group reached (7.48 ± 2.19) scores.After intervened by DEX,NDS of rats in DEX group decreased to (3.15 ± 1.93) scores,significantly lower than in ICH group,the difference was significant (P ＜ 0.05).The necrotic neurons were found around the hematoma of rats in ICH group,while in DEX group,less necrotic neurons were found.(2)In ICH group,the GC level in cerebral homogenate climbed up to a peak of (1.359 1 ±0.308 5) μg/L at 12 h,and slowly went down.By the end of 72 h,the GC level was (0.951 0 ±0.036 1) μg/L,which was higher than those of the CON group[(0.621 3 ±0.039 3) μg/L],the difference was significant (P ＜ 0.05),while in the DEX group,the level of GC in cerebral homogenate showed no difference with statistics from CON group.(3)The mean integrated optical density (IOD) of GR in hippocampal CA1 of rats in the ICH group (1.282 4 ± 0.035 6) were much more smaller than those in the CON group (1.012 5 ± 0.027 3,P ＜ 0.05),which meant the down-regulated expression of GR.(4) No difference was found in the NDS,pathological change,GC level and GR expression between RU486 group and ICH group.DEX didn't effect the expression of GR.Conclusions ICH in neonatal rat disturbs the modulation of hypothalamus-pituitary-adrenal axis,with an increase in the GC level and less GR expression.Early application of exogenous GC helps protect the neurons.%目的 研究发育期大鼠脑出血(ICH)后内源性糖皮质激素(GC)的变化及地塞米松(DEX)的干预效应.方法 新生10日龄SD大鼠按随机数字表法随机分为11组(每组8只):正常对照(CON)组、假手术(SHAM)组、ICH组(每组均按术后时间分别再分为12 h组、24 h组及72 h组),另设糖皮质激素受体(GR)激动剂干预组(DEX组)和GR阻滞剂干预组(RU486组).采用自体血注入法制造ICH模型.DEX组及RU486组大鼠在造模后分别予DEX(1 mg/kg)或RU486 (20 mg/kg)腹腔内注射,SHAM组、CON组及ICH组大鼠仅予9 g/L盐水腹腔内注射.采用神经损伤评分(NDS)进行行为测评.采用放射免疫法测定大鼠脑组织匀浆GC水平.尼氏染色及免疫荧光染色观察神经元形态学变化及海马区GR表达.结果 1.术后72 h ICH大鼠NDS得分升高[(7.48±2.19)分],DEX干预后,NDS得分下降[(3.15±1.93)分],与ICH组比较差异有统计学意义(P＜0.05).ICH组大鼠出血灶周边可见坏死细胞,DEX干预后,血肿周围坏死细胞少.2.ICH组大鼠组织脑匀浆GC水平较SHAM组及CON组明显升高,表现为出血后12 h升高至峰值[(1.359 1 ±0.308 5) μg/L],后逐渐下降[24 h为(1.062 5±0.117 0)μg/L],但在72 h[(0.951 0±0.036 1)μg/L]仍高于CON组[(0.621 3±0.039 3)μg/L,P＜0.05];DEX干预后,脑组织匀浆GC在出血后72 h降至正常水平,与CON组比较差异无统计学意义(P＞0.05).3.ICH组海马CA1区神经元GR平均吸光度值为1.282 4±0.035 6,明显小于CON组(1.0125±0.027 3),2组比较差异有统计学意义(P＜0.05),提示脑出血后GR表达减少.4.RU486干预对其行为学、病理形态、GC分泌及GR表达不产生影响,DEX干预不改变GR的表达.结论 新生大鼠ICH可导致下丘脑-垂体-肾上腺轴失调,表现为GC分泌增多,GR表达减少;早期短程应用DEX可能起神经保护作用.

摘要： Objective To explore the effect of exogenous recombinant human erythropoietin (rhEPO) on neuronal apoptosis in neonatal rats after hyperoxia brain injury.Methods Thirty neonatal Wistar rats were randomly divided into 3 groups by random number table method:rhEPO treatment + 800 mL/L hyperoxia group (group A),9 g/L saline +800 mL/L hyperoxia group (group B),9 g/L saline + air group (group C).Group A was given subcutaneous injection of rhEPO 1 000 IU/kg for 5 days.Group B and group C received the same dose of 9 g/L saline.Group A and group B were continuously exposed to atmospheric pressure hyperoxia model cabin to maintain the oxygen concentration in the container (800 ± 30) mL/L for 5 days.During the course of the experiment,the general situation and weight changes in rats were observed.After 5 d,all rats were sacrificed and brain tissues were taken.Neuronal apoptosis in hippocampal structural region of the newborn rats was observed by terminal deoxynucleotidyl transferase dUTP nick and labeling(TUNEL) staining.Immunohistochemical method was used to detect the expression of 5-lipoxygenase in hippocampal structural region of newborn rats.Results The weight gain and brain weight of group B were lower than those of group C,the weight gain and brain weight of group A were higher than those of group B,and the differences were statistically significant(F =11.179,8.140,all P ＜ 0.05).In group A and group B were found that the neuronal nucleus of the hippocampal neurons was partially contracted,deeply dyed,and the neuronal arrangement was loose,even with local neuron deletions and focal necrosis,but in group A neuron density was higher with less necrosis than that in group B.The neuronal cells in hippocampal structural region were neat and intact in group C.The number of TUNEL positive cells in hippocampal structural region of group B[(6.20 ± 1.93) number/high power field] was significantly higher than that in group C [(1.80 ± 0.79) number/high power field],the number of TUNEL positive cells in hippocampal structural region of group A [(4.20 ± 1.32) number/high power field] was significantly lower than that in group B,and the difference was statistically significant (F =23.912,P ＜ 0.05).The number of 5-lipoxygenase positive cells in group B [(6.90 ± 1.29) number/high power field] was significantly higher than that in group C [(1.00 ± 0.67) number/high power field],the number of 5-lipoxygenase positive cells in group A [(5.60 ± 0.97)number/high power field] was significantly lower than that in group B,and the difference was statistically significant (F =95.044,P ＜ 0.05).Conclusion rhEPO has a protective effect on neonatal rats with hyperoxia brain injury,and alleviates brain cell apoptosis caused by hyperoxia brain injury,which may interfere with the 5-lipoxygenase pathway.%目的 探讨外源性重组人促红细胞生成素(rhEPO)对新生大鼠高氧脑损伤后神经细胞凋亡的影响.方法 将30只新生12 h内的Wistar大鼠按随机数字表法随机分为3组:rhEPO治疗+800 mL/L高氧组(A组),9 g/L盐水+800 mL/L高氧组(B组),9 g/L盐水+空气组(C组).A组每天给予颈后皮下注射rhEPO1 000 IU/kg,共5d.B组和C组给予同样剂量9 g/L盐水.A组和B组持续暴露于常压高氧模型舱中,维持容器内氧体积浓度为(800±30) mL/L,共5d.实验过程中观察大鼠一般情况、体质量改变.5d后处死全部大鼠,取脑组织,采用脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL法)染色检测海马结构区细胞凋亡情况,免疫组织化学法测定海马结构区细胞5-脂氧酶表达.结果 B组体质量增长和脑质量均低于C组,A组体质量增长和脑质量高于B组,差异均有统计学意义(F=11.179、8.140,均P＜0.05).A组和B组发现海马结构区神经元细胞核部分固缩、深染,神经元排列松散,甚至局部神经元缺失、局灶性坏死,但A组较B组神经元密度大,坏死较少;C组海马结构区神经元细胞排列整齐,结构完整.B组海马结构区TUNEL染色阳性细胞数[(6.20±1.93)个/高倍视野]明显多于C组[(1.80±0.79)个/高倍视野],A组海马结构区TUNEL染色阳性细胞数[(4.20±1.32)个/高倍视野]明显少于B组,差异有统计学意义(F=23.912,P＜0.05).B组5-脂氧酶表达阳性细胞数[(6.90±1.29)个/高倍视野]明显多于C组[(1.00±0.67)个/高倍视野],A组5-脂氧酶表达阳性细胞数[(5.60±0.97)个/高倍视野]少于B组,差异有统计学意义(F =95.044,P＜0.05).结论 rhEPO对新生大鼠高氧脑损伤具有脑保护作用,且可能通过干预5-脂氧酶途径减轻高氧脑损伤引起的脑细胞凋亡.

摘要： 目的 探讨腺病毒介导热休克蛋白70 (HSP70)对缺氧性肺动脉高压(HPH)新生大鼠肺的保护作用.方法 选取7 ～10日龄健康、清洁级Wistar新生大鼠128只,按随机数字表法随机分为HPH组和对照组.HPH组根据转染液不同分为盐水组、空病毒组、HSP70组,转染后置于80 mL/L氮氧混合气体的低氧舱内建立HPH模型.造模3、7、10、14 d测定各组新生大鼠平均肺动脉压力(mPAP).应用反转录PCR和Western blot检测各组新生大鼠肺组织中HSP70、缺氧诱导因子-1α(HIF-1α)、内皮素-1(ET-1)、诱导型一氧化氮合酶(iNOS) mRNA及蛋白表达.结果 1.缺氧3、7、10、14 d盐水组mPAP水平(M,Q:12.00,2.50;15.00,2.00;18.00,1.75;20.00,2.25)与对照组(M,Q:9.50,4.75;10.50,1.00;13.00,1.00;15.50,3.25)比较显著增高,差异均有统计学意义(z=-3.28、-3.40、-3.34、-3.06,均P＜0.01);空病毒组(M,Q:13.50,2.00;15.50,1.75;18.00,1.00;22.00,4.25)与对照组比较显著增高,差异均有统计学意义(z=-2.83、-3.40、-3.42、-2.97,均P＜0.01);HSP70组缺氧3、7、10 d的mPAP水平(M,Q:8.50,4.00;10.50,1.00;13.00,1.oo)与对照组比较差异均无统计学意义(z=-0.43、-0.00、-3.06,均P＞0.05).2.HSP70组缺氧3、7、10 d各时间点间HIF-1 α、ET-1、iNOS mRNA比较差异均有统计学意义(F=6.321、9.669、6.333,均P＜0.01),HSP70蛋白比较差异均有统计学意义(F=16.463、3.637、17.749,均P＜0.01).3.缺氧3、7、10 d盐水组HIF-1α mRNA显著高于对照组,差异均有统计学意义(q =4.312、9.106、6.151,均P＜0.01);空病毒组显著高于对照组,差异均有统计学意义(q=3.982、9.235、5.352,均P＜0.01);缺氧3、7 d HSP70组低于空病毒组,差异均有统计学意义(q=6.083、11.031,均P＜0.05).缺氧3、7、10 d盐水组ET-1 mRNA显著高于对照组,差异均有统计学意义(g=5.112、10.086、6.264,均P＜0.01);空病毒组显著高于对照组,差异均有统计学意义(q =4.182、12.238、5.864,均P＜0.叭);缺氧3、7、10 d HSP70组低于空病毒组,差异均有统计学意义(g=6.912、10.235、7.021,均P＜0.05).缺氧3、7、10 d盐水组iNOS mRNA显著高于对照组,差异均有统计学意义(q=4.998、8.056、5.369,均P＜0.01);空病毒组显著高于对照组,差异均有统计学意义(q=4.778、10.138、5.154,均P＜0.01),缺氧3、7、10 d HSP70组低于空病毒组,差异均有统计学意义(q=7.819、9.838、6.156,均P＜0.05).缺氧3、7、10 d盐水组HIF-1 α蛋白显著高于对照组,差异均有统计学意义(q=3.146、3.012、4.106,均P＜0.05);缺氧10 d空病毒组显著高于对照组,差异有统计学意义(q =3.468,P＜0.05);缺氧3、7、10 d HSP70组显著低于空病毒组,差异均有统计学意义(q=3.876、4.108、4.021,均P＜0.05).缺氧3、7、10 d HSP70组ET-1蛋白显著低于盐水组,差异均有统计学意义(q=3.367、2.983、3.246,均P＜0.05);且显著低于空病毒组,差异均有统计学意义(q=3.268、2.678、3.567,均P＜0.05).缺氧3、7、10 d盐水组iNOS蛋白显著高于对照组,差异均有统计学意义(q=3.360、3.567、3.567,均P＜0.05),HSP70组低于空病毒组,差异均有统计学意义(q=3.126、3.908、3.087,均P＜0.05).结论 腺病毒介导HSP70可以提高HPH新生大鼠肺组织HSP70表达,下调HIF-1α、ET-1、iNOS表达,降低肺动脉压力.%Objective To investigate the protective effect of adenovirus mediated heat shock protein 70 (HSP70) on lungs in neonatal rats with hypoxic pulmonary hypertension (HPH).Methods One hundred and twenty-eight 7-10 d healthy Wistar neonatal rats were randomly divided into HPH model group and control group.HPH group was divided into saline group,empty virus group,and HSP70 group according to the transfection solution.HPH model was established in the hypoxia cabin of 80 mL/L nitrogen oxygen mixed gas after transfection.The mean pulmonary artery pressure(mPAP) was measured after 3,7,10 and 14 days of hypoxia in each group.The mRNA and protein expression of HSP70,hypoxia inducible factor-1 alpha(HIF-1 α),endothelin-1 (ET-1) and inducible nitric oxide synthase(iNOS) in the lung tissues of neonatal rats were detected by using reverse transcription-PCR and Western blot respectively.Results (1) The mPAP level was significantly higher in saline group (M,Q:12.00,2.50;15.00,2.00;18.00,1.75;20.00,2.25) than that in control group (M,Q:9.50,4.75;10.50,1.00;13.00,1.00;15.50,3.25),and the differences were significant (z =-3.28,-3.40,-3.34,-3.06,all P ＜ 0.01);and the differences were also significant between empty virus group (M,Q:13.50,2.00;15.50,1.75;18.00,1.00;22.00,4.25) and control group (z =-2.83,-3.42,-3.40,-2.97,all P ＜ 0.01) in 3,7,10,and 14 days;but there was no significant difference between HSP70 group (M,Q:8.50,4.00;10.50,1.00;13.00,1.00)and the control group in 3,7,and 10 days (z =-0.43-0.00,-3.06,all P ＞ 0.05).(2) The expressions of HSP70 mRNA among the groups were statistically significant(F =6.321,9.669,6.333,all P ＜ 0.01),and the expressions of HSP70 protein also had significant difference(F =16.463,3.637,17.749,all P ＜ 0.01).(3)The level of HIF-1α mRNA in saline group was significantly higher than that of the control group,and the differences were statistically significant (q =4.312,9.106,6.151,all P ＜ 0.01);and the level of HIF-1α mRNA in empty virus group was also significantly higher than that in the control group,and the differences were statistically significant (q =3.982,9.235,5.352,all P ＜ 0.01) in 3,7,and 10 days;hypoxia in HSP70 group was lower than that of the empty virus group in 3,7 days,and the differences were statistically significant (q =6.083,11.031,all P ＜ 0.05).The level of ET-1 mRNA in saline group was significantly higher than that in the control group(q =5.112,10.086,6.264,all P ＜ 0.01),in empty virus group was significantly higher than that in the control group,and the differences were statistically significant (q =4.182,12.238,5.864,all P＜0.01) in 3,7,and 10 days,but in HSP70 group it was lower than that in the empty virus group in 3,7,and 10 days,and the differences were statistically significant (q =6.912,10.235,7.021,all P ＜ 0.05).The level of iNOS mRNA in saline group was significantly higher than that of the control group,and the differences were statistically significant (q =4.998,8.056,5.369,all P ＜0.01),in empty virus group was significantly higher than that in the control group,and the differences were statistically significant (q =4.778,10.138,5.154,all P ＜0.01) in 3,7,and 10 days,but in HSP70 group it was lower than that in the empty virus group in 3,7,and 10 days,and the differences were statistically significant (q =7.819,9.838,6.156,all P ＜ 0.05).The level of HIF-1 α protein in saline group was significantly higher than that of the control group in 3,7,and 10 days,and the differences were statistically significant (q =3.146,3.012,4.106,all P ＜ 0.05),in empty virus group was significantly higher than that of the control group in 10 days,and the difference was statistically significant (q =3.468,P ＜ 0.05);but in HSP70 group it was lower than that in the empty virus group in 3,7,and 10 days,and the differences were statistically significant (q =3.876,4.108,4.021,all P＜ 0.05).The level of ET-1 protein of HSP70 group was lower than that of the saline group,the differences were statistically significant(q =3.367,2.983,3.246,all P ＜ 0.05),in HSP70 group was lower than that of the empty virus,and the differences were statistically significant (q =3.268,2.678,3.567,all P ＜0.05).The level of iNOS protein in saline group was significantly higher than that in the control group in 3,7,and 10 days,and the differences were statistically significant (q =3.360,3.567,3.567,all P ＜ 0.05),but in HSP70 group it was lower than that in the empty virus group,and the differences were statistically significant (q =3.126,3.908,3.087,all P ＜ 0.05).Conclusion Adenovirus mediated HSP70 can improve the HSP70 expression in HPH,down-regulate the expression of HIF-1 α,ET-1,iNOS,and reduce pulmonary arterial pressure.

摘要： 目的 观察新生大鼠缺氧缺血性脑损伤(HIBD)模型的AMP-活化蛋白激酶(AMPK)、核转录因子FOXO3a及凋亡蛋白活化型天冬氨酸特异性半胱氨酸蛋白酶3(CC3)表达水平变化,探讨抑制AMPK/FOXO3a信号通路对新生大鼠缺氧缺血神经元凋亡的保护作用机制.方法 将64只7 d龄SD大鼠按照随机数字表法分为4组:缺氧缺血组(HI组)、假手术组(sham组)、AMPK特异性抑制剂Compound C干预组(干预组)、二甲基亚砜(DMSO)溶剂组(对照组),每组SD大鼠各为16只.建模方法:对HI组SD大鼠在乙醚麻醉下进行右侧颈总动脉结扎,采用氧、氮混合气体(8％ O2、92％ N2)缺氧处理2.5 h;对sham组SD大鼠仅分离右侧颈总动脉,不结扎,不进行缺氧处理;对干预组和对照组SD大鼠分别于右侧脑室内注射等量Compound C和DMSO,30 min后,采用氧、氮混合气体(8％ O2、92％ N2)进行缺氧处理.分别于建模24 h后处死4组SD大鼠,取大脑皮质,采用Western印迹法定量检测各组SD大鼠模型大脑皮质的AMPK蛋白、p-AMPKα蛋白、总FOXO3a蛋白、细胞核FOXO3a蛋白、细胞质FOXO3a蛋白及CC3表达水平;应用原位缺口末端标记(TUNEL)染色法,检测凋亡神经元细胞.结果 ①与sham组相比,HI组及对照组SD大鼠模型大脑皮质的AMPK蛋白水平与总FOXO3a蛋白水平均无显著变化,并且差异均无统计学意义(P>0.05);p-AMPKα蛋白水平及细胞核FOXO3a蛋白水平均显著增高,细胞质FOXO3a蛋白水平均显著降低,CC3表达水平均显著增高,并且差异均有统计学意义(P0.05).However, expression levels of p-AMPKα protein and nuclear protein of FOXO3a in HI group and control group all were much higher than those in sham group, and the cytoplasmic protein of FOXO3a were evidently lower than that in sham group, which up-regulated expression level of CC3 protein after hypoxic-ischemic in HI group and control group compared with sham group, and all the differences were statistically significant (P<0.01).②Expression levels of p-AMPKα protein and nuclear protein of FOXO3a in treated group were obviously lower than those in HI group and control group, and expression level of cytoplasmic protein was much higher than those in HI group and control group, and meanwhile leading to the decrease of CC3 protein after hypoxic-ischemic in treated group compared with HI group and control group, and all the differences were statistically significant (P<0.01).③Apoptosis levels of neuronal cell (expression levels of TUNEL positive cells) in HI group and control group both were higher than that in sham group, and both the differences were statistically significant (P<0.01).TUNEL positive cells were obviously reduced in treated group compared with those in HI group and control group, and all the differences were statistically significant (P<0.01).Conclusions AMPK activity increased in the neonate rat brain with HIBD.AMPK activity inhibition can inhibit FOXO3a translocation from cytoplasm to nucleus, down-regulate the level of CC3 protein, leading to the reduction of neuronal apoptosis.

摘要： Objective To investigate the expression of long non - coding RNA(lncRNA)in neonatal rats with hypoxic - ischemic brain damage(HIBD). Methods SD rats of 10 postnatal days were divided into the sham -operated control and the hypoxic - ischemic(HI)group. At 24 h after HI,the animals were sacrificed. HE staining was used to assess histopathological damage. Microarray was used to detect the expression of lncRNA and mRNA in hypoxic -ischemic and sham control brain. Real - time PCR was used to verify the microarray result. The differentially expressed mRNA was analyzed by gene ontology(GO),pathway and coding - noncoding RNA co - expression(CNC)network analysis. Results HE staining showed that cells in HI brains became swollen and disordered with ambiguous cell struc-ture. Microarray data demonstrated that 322 lncRNAs and 375 mRNAs were significantly altered in the neonatal brains following hypoxic - ischemic injury compared with sham control(P ﹤ 0. 05). The real - time PCR results agreed with those of the microarray. GO analysis showed that the most enriched biological process associated with the upregulated mRNA had response to wounding,whereas the biological process mostly enriched among the downregulated mRNA was so-matic stem cell division. Pathway analysis indicated that upregulated mRNA was primarily corresponded with cytokine -cytokine receptor interaction pathway and that downregulated mRNA mainly correlated to axon guidance pathway. CNC network analysis demonstrated that 177 lncRNAs were correlated to the expression of mRNA involved in inflammation and cell death(P ﹤0. 05). Conclusions HI injury significantly influences cerebral lncRNA and mRNA expression profiles in the neonatal rat brains. Deregulated lncRNAs might contribute to the pathogenesis of HIBD via interacting with mRNA.%目的：探讨长链非编码 RNA（lncRNA）在新生大鼠缺氧缺血性脑损伤中的表达情况。方法10日龄 SD 大鼠分为假手术组和缺氧缺血组，于缺氧缺血24 h 处死取脑。采用 HE 染色观察脑组织病理改变，采用芯片技术检测缺氧缺血组和假手术组 lncRNA 和 mRNA 的表达，采用实时定量 PCR 验证芯片数据。对差异表达 mRNA 进行基因本体论（GO）分析、通路分析和编码-非编码基因共表达网络等生物信息学分析。结果HE 染色显示缺氧缺血组脑组织细胞肿胀、结构不清，排列紊乱。芯片数据表明，与假手术组相比，缺氧缺血组脑组织中322个 lncRNA 和375个 mRNA 呈差异表达（P ﹤0.05），实时定量 PCR 结果与芯片结果一致。GO 分析结果表明，上调表达的 mRNA 主要富集于损伤反应过程，而下调表达的 mRNA 则主要富集于成体干细胞分裂过程。通路分析结果表明，上调表达的 mRNA 主要与细胞因子及其受体相互作用通路相关，而下调表达mRNA 则主要与轴突导向通路相关。编码-非编码基因共表达网络分析发现，177个 lncRNA 与炎症、细胞死亡方面的 mRNA 表达相关（ P ﹤0.05）。结论缺氧缺血导致新生鼠脑组织中 lncRNA 和 mRNA 显著改变， lncRNA 可能通过与 mRNA 相互作用参与发育期脑缺氧缺血损伤的病理过程。

摘要： 本发明公开了一种新生大鼠海马神经元原代培养的培养液及原代培养大鼠海马神经元的方法，该培养液是在基础培养液Neurobasalmedium中加入1、6二磷酸果糖、果糖和营养添加剂。原代培养新生大鼠海马神经元具有以下步骤：①制备海马神经元培养液；②取材、消化；③制备单细胞悬液；④接种、培养。本发明原代培养大鼠培养海马神经元的方法简便，采用本发明培养液能够获得生长状态良好、纯度较高的海马神经元，具有重要的应用价值和经济价值。

摘要： 本发明涉及生物技术领域，更具体地涉及分离原代新生大鼠心肌细胞(NRCM)的方法，所述方法采用两种消化酶进行逐步消化，并且不包括离心步骤。

摘要： 本发明属于动物细胞培养技术领域，涉及一种新生大鼠皮肤终隔细胞大量扩增方法及应用。步骤如下：新生大鼠皮肤样品的预处理；酶消化液配置；酶消化液处理样品，获得终隔细胞悬液；制备新生大鼠皮肤终隔细胞原代细胞；原代细胞的传代扩大培养，制备传代细胞；c‑kit+/CD34+流式细胞仪分选纯化皮肤终隔细胞，重悬细胞液，扩大培养，制备纯度较高的皮肤终隔细胞。本发明采用0.05%胶原酶II、0.05%胰蛋白酶，联合消化法分离培养皮肤终隔细胞，该方法得到的原代终隔细胞数量大，纯度高，活力好，经流式细胞仪分选后，可快速大量扩增，扩增后细胞形态稳定，活力高，可广泛用于后续相关终隔细胞形态检测和功能研究。

摘要： 一种研究高压氧对脑出血大鼠脑内血管新生影响的方法，将SD大鼠随机分为假手术组(A组)、模型组（B组）和高压氧组（C组）。采用组织切片HE染色显微镜下观察各组大鼠血肿周围脑组织新生血管形成情况；采用免疫组织化学分析方法检测各组大鼠脑内HIF1-α、VEGF的蛋白表达含量；采用荧光定量RT-PCR方法测定各组大鼠脑组织HIF1-αmRNA、VEGFmRNA的表达水平。

摘要： 本发明提出一种新生大鼠嗅球嗅鞘细胞(OECs)的分组纯化方法，其应用于新生大鼠嗅球嗅鞘细胞的纯化过程，所述纯化过程包括OECs的取材；OECs的分离：OECs分组纯化及比较；形态学观察以及OECs纯度检测。本发明通过不同的的新生大鼠嗅球嗅鞘细胞(OECs)的纯化方法实验，结果显示体外培养的新生大鼠嗅球OECs主要为双极或三级细胞，其突起细长。未经纯化的OECs则成纤维细胞生长迅速而占优势，三种纯化方法均能使接种14d后的OECs纯度均值大于75％。p75抗体吸附法和阿糖胞苷抑制法的纯化率均值略高于差速贴壁法。故新生大鼠嗅球OECS的原代培养中细胞纯化是必要的；在脊髓损伤再生研究中，较之p75抗体吸附法和阿糖胞苷抑制法，差速贴壁法是一种简单、经济、实用的OECs纯化方法。

摘要： 本发明公开了一种新生大鼠海马神经元原代培养的培养液及原代培养大鼠海马神经元的方法，该培养液是在基础培养液Neurobasalmedium中加入1、6二磷酸果糖、果糖和营养添加剂。原代培养新生大鼠海马神经元具有以下步骤：①制备海马神经元培养液；②取材、消化；③制备单细胞悬液；④接种、培养。本发明原代培养大鼠培养海马神经元的方法简便，采用本发明培养液能够获得生长状态良好、纯度较高的海马神经元，具有重要的应用价值和经济价值。

摘要： 本发明提出一种新生大鼠嗅球嗅鞘细胞(OECs)的分离方法，其应用于OECs的纯化过程，所述纯化过程包括OECs的取材；OECs的分离：OECs分组纯化及比较；形态学观察以及OECs纯度检测。本发明通过不同的新生大鼠嗅球嗅鞘细胞(OECs)的纯化方法实验，结果显示体外培养的新生大鼠嗅球OECs主要为双极或三级细胞，其突起细长。未经纯化的OECs则成纤维细胞生长迅速而占优势，三种纯化方法均能使接种14d后的OECs纯度均值大于75％。p75抗体吸附法和阿糖胞苷抑制法的纯化率均值略高于差速贴壁法。故新生大鼠嗅球OECS的原代培养中细胞纯化是必要的；在脊髓损伤再生研究中，较之p75抗体吸附法和阿糖胞苷抑制法，差速贴壁法是一种简单、经济、实用的OECs纯化方法。

摘要： 本发明提供一种在大鼠离乳期给与3‑MA处理制备神经发育缺陷大鼠模型的方法，包括给予正常离乳期大鼠连续7天腹腔注射自噬抑制剂3‑MA制备抑制自噬发生的大鼠模型，并通过免疫印迹检测自噬发生，而后通过三箱社交实验验证社交障碍、开放旷场实验、高架十字迷宫实验以及黑白箱实验验证大鼠模型焦虑水平是否增加，通过Y迷宫实验和被动规避实验验证大鼠模型是否出现记忆损伤，大鼠模型与对照组进行高尔基体染色，观察海马和额叶神经元及突触形态。通过多次实验发现，离乳期3‑MA处理制备神经发育缺陷的大鼠模型，具有成本低廉，造模周期短，模型行为异常更明显，稳定性更好的优势，可用于精神分裂症、孤独症等疾病研究及新药研发。

摘要： 一种新生大鼠手术台，其包括：固定大鼠的台体；台体的周缘设有防止大鼠逃脱固定的板体，且板体与台体可合围成一盒体；在台体上设置一可绕台体平面一直线翻折的展示板，且对应大鼠躯干及四肢位置在展示板上分别设置固定大鼠躯干及四肢的束紧带；束紧带可在展示板上进行移动；使用时利用板体与台体组成的盒体可以便于工作人员进行大鼠手术台的场地更换，且可将台体作为手术台完全展示在工作人员眼前；利用可活动翻折的展示板可以帮助工作人员观察大鼠后侧躯干；利用可移动的束紧带可以帮助工作人员观察大鼠被固定位置。

摘要： 本实用新型提供一种经玻璃体腔注射大鼠诱导视网膜蜕变用针对大鼠的注射器，涉及针筒技术领域，本实用新型包括本体和固定装置，所述本体为注射器，所述本体的一端连通有连接口，所述连接口远离本体的一端设有针头，所述本体的表面设有固定装置，所述固定装置包括连接板，所述连接板的内部与本体贯穿滑动连接，所述连接板的一侧固定连接有滑柱，所述滑柱远离连接板的一端滑动连接有套环，所述套环的表面固定连接有两个固定板，所述固定板呈“L”形，且固定板的长臂端贯穿插设有圆柱。本实用新型解决了在注射的过程中老鼠可能会乱动，可能会导致人无法一次性的将注射液通过注射器注射进老鼠体内的问题。