SD大鼠乳鼠原代皮质神经元和小胶质细胞同时提取并培养的实验方法

背景:原代皮质神经元和小胶质细胞在探索神经系统疾病细胞疗法中起着至关重要的作用,而目前获得2种细胞的方法大多较繁琐,需分别单独提取。因此找到一种便捷、快速可同时提取2种细胞的方法十分关键。目的:探索一种新型同时提取原代皮质神经元及小胶质细胞的方法。方法:取24 h内新生SD大鼠乳鼠,将大脑取出放入装有DMEM的培养皿中,去除软脑膜后备用。同一脑组织,先提取原代神经元,再将剩余脑组织用于提取小胶质细胞,整个过程在冰上操作。原代皮质神经元提取培养步骤:镊子夹取2.0-3.0 mm厚度大脑皮质组织置于培养皿,用木瓜蛋白酶消化20 min,中止消化后吹打组织呈互相粘连的组织悬液,吸上清细胞悬液,过滤、分装到15 mL离心管中,离心并重悬后将细胞接种于左旋多聚赖氨酸包被的6孔板爬片上放入细胞培养箱,每隔1 d观察神经元形态,第7天进行MAP2和β-Tubulin免疫荧光染色鉴定。小胶质细胞提取培养步骤:夹取上一步取皮质后剩余脑组织,厚度8-10 mm,置于培养皿,用胰酶消化20 min,中止消化后吹打组织呈匀浆,然后将匀浆转移到培养瓶中培养,第14天将培养瓶封口后进行恒温水平摇床振荡2 h,小胶质细胞脱落于上清中;取纯化后的小胶质细胞继续培养3 d进行Iba1免疫荧光染色鉴定。结果与结论:(1)神经元培养24 h后贴壁、胞体变大,部分神经元长出突触;培养到第3,5天时胞体进一步增大,大部分神经元已呈突触形态,部分神经元成团生长;第7天时,神经元突触延长增粗并互相连接成网。经β-Tubulin、MAP2免疫荧光染色鉴定为神经元。(2)小胶质细胞培养第1天换液后可见细胞贴壁生长;第3,5,7天时细胞密度均较小,细胞形态呈高亮椭圆或圆形,但已基本成团块状生长于其他细胞上层;第10天时小胶质细胞密度明显增大;第14天时小胶质细胞呈致密团块状生长于其他细胞上层,此时可进行分离纯化。取分离纯化后细胞再培养至第3天,经Iba1免疫荧光鉴定为小胶质细胞,且纯度大于95%。(3)结果表明,经此法提取并培养获得的原代皮质神经元和小胶质细胞纯度高、形态好、成活率高。

吸入暴露途径下纳米塑料在SD大鼠体内的富集特征研究

为探讨吸入暴露纳米级塑料在生物体呼吸系统以外组织上的富集特征,以100 nm聚苯乙烯微球对SD大鼠进行2 h短期吸入暴露,使用热裂解-气相色谱质谱法测定大鼠肝脏、肾脏、心脏以及脑组织上的聚苯乙烯浓度。结果显示:暴露组大鼠肾脏、心脏和肝脏中的浓度[(0.416±0.143)、(0.633±0.278)和(0.617±0.179)g/g]显著高于对照组大鼠[(0.249±0.020)、(0.070±0.096)和(0.101±0.140)g/g],差异具有统计学意义(P<0.05),但各组织间的相关性不显著(P>0.05);2组大鼠脑组织均未检出聚苯乙烯。研究表明,100 nm聚苯乙烯微球吸入暴露2 h后,可通过肺泡进入血液循环,进而在大鼠肝脏、肾脏、心脏中形成沉积。通过探讨纳米级塑料在生物体内的累积特征,为深入开展吸入纳米塑料对人体的健康影响研究提供数据支撑。

基于液质联用法测定嘧啶核苷类化合物GY005在SD大鼠体内的药代动力学实验研究

建立LC-MS/MS法测定嘧啶核苷类化合物GY005在大鼠血浆中的浓度,并应用于大鼠体内的药代动力学研究。以特非那定为内标,经乙腈-甲醇沉淀蛋白后,采用ACE 5 C4 column(50 mm×2.1 mm,2.5μm)色谱柱,以0.1%甲酸水和0.1%甲酸乙腈为流动相,进行梯度洗脱分离,流速0.7 mL·min^(-1),进样量10μL;采用电喷雾离子源(ESI),正离子扫描方式,MRM扫描,分别以m/z 445.15→270.10和m/z 472.40→436.40作为化合物GY005和内标(特非那定)的质谱检测条件。大鼠灌服2 mg·kg^(-1)化合物GY005后,不同时间点取样测定其血浆浓度,计算药代动力学参数。雌雄SD大鼠单次静脉注射2 mg·kg^(-1)的受试品后,化合物GY005在雄性大鼠体内的暴露量(AUC_(last))为(228±44)h·ng·mL^(-1),消除半衰期(T_(1/2))为(0.30±0.01)h,清除率(CL)为(149±29)mL·min^(-1)·kg^(-1)。化合物GY005在雌性大鼠体内的暴露量(AUC_(last))为(215±158)h·ng·mL^(-1),消除半衰期(T_(1/2))为(0.32±0.03)h,清除率(CL)为(226±162)mL·min^(-1)·kg^(-1)。雌雄间暴露水平相当,没有显著性差异。这个方法具有简便、灵敏、快捷,适用于化合物GY005的药代动力学研究。

菜籽油对SD大鼠糖脂代谢的影响

本实验采用微波对菜籽进行预处理并压榨制成菜籽油(rapeseed oil,RO),探究RO干预后对高糖高脂饮食SD大鼠糖脂代谢的影响。结果表明:RO能够抑制SD大鼠体内HbAlc的上升和促进肝糖原的合成以改善血糖代谢。RO能够显著降低SD大鼠血浆中TC、TG、LDL-C的水平和提高HDL-C以改善血脂代谢,同时提高SD大鼠血浆中GSH-Px、SOD、CAT的活性和GSH含量,并减少降低MDA的生成以降低血糖血脂对脏器的损伤。此外,RO通过降低ApoB含量以及增加ApoA1含量来调控LDL-C和HDL-C。因此,RO可有效地调节血糖血脂代谢、提高血浆抗氧化能力,缓解氧化应激。RO具备开发成为辅助降血糖血脂制品原料的潜力。

刺山柑果风湿止痛凝胶膏对SD大鼠胚胎-胎仔发育的影响

目的:观察刺山柑果风湿止痛凝胶膏对SD大鼠胚胎-胎仔发育的影响。方法:实验设刺山柑果风湿止痛凝胶膏高(117.8 g/m^(2))、中(58.9 g/m^(2))、低(29.5 g/m^(2))剂量组,辅料对照组(辅料帖)和阳性对照组(环磷酰胺),妊娠第6天(GD6)~GD15每天经皮给药1次,阳性对照组于GD12皮下注射环磷酰胺。实验期间观察孕鼠的一般状况、体质量和饲料消耗量;GD20处死孕鼠,取胎仔,检查子宫连胎质量、黄体数、着床数、胎仔质量、胎仔身长、胎仔尾长、活胎数、死胎数、吸收胎数、胎仔性别及外观、内脏和骨骼畸形。结果:刺山柑果风湿止痛凝胶膏各剂量组孕鼠体质量、体质量增量、摄食量、子宫连胎质量、子宫和卵巢的脏器质量及其系数、平均黄体数、着床前丢失率、平均着床数、着床率、平均活胎数、活胎率、死胎率、吸收胎率、胎仔性别比、胎仔的外观异常率、窝均体质量、窝均身长、窝均尾长、骨骼畸形率和内脏畸形率与辅料对照组相比差异均无统计学意义(均为P>0.05)。结论:刺山柑果风湿止痛凝胶膏对SD大鼠胚胎-胎仔发育毒性的未见不良反应水平为117.8 g/m^(2),是临床拟用体表剂量的12倍。在本试验条件下,SD大鼠经皮给予刺山柑果风湿止痛凝胶膏对胚胎-胎仔发育无毒性影响。

Effect of transforming growth factor beta and bone morphogenetic proteins on rat hepatic stellate cell proliferation and transdifferentiation

AIM: To explore different roles of TGF-β (transforming growth factor beta) and bone morphogenetic proteins (BMPs)in hepatic stellate cell proliferation and trans-differentiation.METHODS: Hepatic stellate cells were isolated from male Sprague-Dawley rats. Sub-cultured hepatic stellate cells were employed for cell proliferation assay with WST-1 reagent and Western blot analysis with antibody against smooth muscle alpha actin (SMA).RESULTS: The results indicated that TGF-β1 significantly inhibited cell proliferation at concentration as low as 0.1 ng/ml, but both BMP-2 and BMP-4 did not affect cell proliferation at concentration as high as 10 ng/ml. The effect on hepatic stellate cell trans-differentiation was similar between TGFβ1 and BMPs. However, BMPs was more potent at transdifferentiation of hepatic stellate cells than TGF-β1. In addition, we observed that TGF-β1 transient reduced the abundance of SMA in hepatic stellate cells.CONCLUSION: TGF-β may be more important in regulation of hepatic stellate cell proliferation while BMPs may be the major cytokines regulating hepatic stellate cell transdifferentiation.

Magnetic resonance imaging of hepatocellular carcinoma induced by diethylnitrosamine in Sprague-Dawley rats

Diethylnitrosamine (DENA) is able to induce various benign and malignant liver lesions in rats with a high success rate and a low mortality rate. It provides a more appropriate model that better simulates the various lesions occurring in humans than the usual model of tumor implantations. The aim of the present study was to evaluate MRI liver examination in Sprague-Dawley(SD) rats as a routine method to detect hepatocellular carcinoma (HCC) nodules induced by DENA and to follow up their growth. METHODS:Hepatic carcinogenesis was induced in 80 male SD rats using oral DENA solution. All animals were imaged for liver tumor detection with a 1.5 Tesla magnet (Siemens Sonata,Erlangen, Germany) using correspondence scan parameters and a radio-frequency knee coil. Macroscopic examinations were performed along the axial MRI sections to evaluate magnetic resonance imaging (MRI) findings, and histopathological assessment was also performed. RESULTS:No false negative results were obtained on MR images. Hepatic tumors in 72 rats were confirmed macro-scopically and 68 rats were detected by MRI till the 20th week. The smallest and the largest nodules detected by MRI were 2 mm and 37 mm in diameter respectively. The agreement rate of MRI with macroscopic observation was 39. 1% and 97. 4% respectively for 2 mm to 5 mm and more than 5 mm nodules. CONCLUSIONS:The hepatic tumor induced by DENA provides a more representative range of tumors for imaging diagnosis and interventional treatment. MRI is the best approach for scrutinizing pathological changes of rat livers in the period of observation.

Toxic Effects of Tetrabromobisphenol A on Thyroid Hormones in SD Rats and the Derived-reference Dose

The present study determined the thyroid hormone interference of tetrabromobisphenol A （TBBPA） in Sprague-Dawley （SD） rats, and the derived-reference dose （RfD） of different endpoint effects on mammals based on experimental results and data collection. Based on repeated exposure toxicity tests on mammals and extensive research, the present study used BMDS240 Software to derive a benchmark dose, and analyzed the accuracy and uncertainty, and similarity with other studies. Test results on triiodothyronine （T3）, thyroxine （T4）, and thyroid stimulating hormone （TSH） demonstrated that all the indicators presented a non-monotonous dose-effect relationship clearly, except TSH in male rats exposed to 0-1000 mg/kg BW per day. Therefore, RfDs were derived from different critical effects. In summary, RfD for mammals in the present study was found to be 0.6 mg/kg per day.

Analysis of transcriptome sequencing of sciatic nerves in Sprague-Dawley rats of different ages

An aging-induced decrease in Schwann cell viability can affect regeneration following peripheral nerve injury in mammals. It is therefore necessary to investigate possible age-related changes in gene expression that may affect the biological function of peripheral nerves. Ten 1-week-old and ten 12-month-old healthy male Sprague-Dawley rats were divided into young（1 week old） and adult（12 months old） groups according to their ages. mRNA expression in the sciatic nerve was compared between young and adult rats using next-generation sequencing（NGS） and bioinformatics（n = 4/group）. The 18 groups of differentially expressed mRNA（DEmRNAs） were also tested by quantitative reverse transcription polymerase chain reaction（n = 6/group）. Results revealed that（1） compared with young rats, adult rats had 3608 groups of DEmRNAs. Of these, 2684 were groups of upregulated genes, and 924 were groups of downregulated genes. Their functions mainly involved cell viability, proliferation, differentiation, regeneration, and myelination.（2） The gene with the most obvious increase of all DEmRNAs in adult rats was Thrsp（log2 FC = 9.01, P 〈 0.05）, and the gene with the most obvious reduction was Col2 a1（log2 FC = -8.89, P 〈 0.05）.（3） Gene Ontology analysis showed that DEmRNAs were mainly concentrated in oligosaccharide binding, nucleotide-binding oligomerization domain containing one signaling pathway, and peptide-transporting ATPase activity.（4） Analysis using the Kyoto Encyclopedia of Genes and Genomes showed that, with increased age, DEmRNAs were mainly enriched in steroid biosynthesis, Staphylococcus aureus infection, and graft-versus-host disease.（5） Spearman＇s correlation coefficient method for evaluating NGS accuracy showed that the NGS results and quantitative reverse transcription polymerase chain reaction results were positively correlated（rs = 0.74, P 〈 0.05）. These findings confirm a difference in sciatic nerve gene expression between adult and young rats, suggesting that, in peripheral nerves, cells and the microenvironment change with age, thus influencing the function and repair of peripheral nerves.

Hormetic Effects of Yttrium on Male Sprague-Dawley Rats

To evaluate hormesis induced by Yttrium（Y） nitrate in male rats, Y was offered to F0 mother rats and F1 offspring at concentrations of 0, 20, 80, and320 ppm daily from gestational day （GD） 0 through postnatal day 70（PND70）.The F1 offspring were evaluated with respect to motor function,learning and memory, and histopathology.

SD大鼠胚胎-胎仔发育毒性试验背景数据的建立

目的总结国家药物安全评价监测中心2013-2022年SD大鼠胚胎-胎仔发育毒性试验各指标的正常值范围,建立SD大鼠胚胎-胎仔发育毒性试验各指标的背景数据库,为药物胚胎-胎仔发育毒性评价提供参考。方法对本中心2013-2022年11项SD大鼠胚胎-胎仔发育毒性试验中对照组共计205只孕鼠和3037只胎鼠各项胚胎发育和胎仔生长发育指标进行统计分析,计算其平均数、标准差、变异系数和95%置信区间。指标包括孕鼠妊娠期体重和体重增长幅度、孕鼠摄食量、妊娠结局(妊娠率、平均黄体数、平均着床数、平均活胎数、活胎率、吸收胎率、死胎率)、胎仔生长发育情况(胎仔重、胎盘重、性别比)、胎仔外观异常率、内脏异常率和骨骼异常率。结果孕鼠妊娠期体重呈增长趋势,妊娠后期体重增长幅度明显增大。孕鼠摄食量随着妊娠时间的增加呈增长趋势。妊娠第20天进行剖腹产,妊娠率为93.2%,平均黄体数、着床数和活胎数分别为18.0±3.2,15.9±2.8和14.8±3.0,活胎率为93.4%,死胎率为6.6%;胎仔雄/雌性别比为0.94,平均体重为(3.6±0.3)g,胎盘平均重量为(0.6±0.3)g。胎仔外观异常发生率约为0.2%,内脏异常率约为0.8%。骨骼异常率约为1.2%,未骨化和骨化不全的发生率较高,主要发生于胸骨和舌骨等,胎仔的掌骨骨化数、跖骨骨化数和骶尾椎骨化数分别为7.0±0.7,8.0±0.1和7.4±0.5,第Ⅰ~Ⅳ胸骨骨化率较高,平均为98.6%~99.9%,第Ⅴ胸骨骨化率为(68.0±28.4)%,第Ⅵ胸骨骨化率为(82.8±23.9)%。结论初步建立了本GLP实验室SD大鼠胚胎-胎仔发育毒性试验中各指标背景数据库,为生殖毒性研究提供参考。

Comparative study on the difference of obesity model induced by two kinds of high fat diet in Sprague-Dawley rats

Objective: To explore the differences of obese Sprague-Dawley(SD)rats model induced by lard oil high-fat(HF)diet or purified HF diet. Methods: SD weanling rats were randomly divided into three groups: D1 group,where rats were fed by lard oil HF diet;D2 group,where rats were fed by purified HF diet;C group,where rats were fed on chow. After 12 weeks,diet-induced obesity rat(stop 33% based on weight)were selected for further study,and the rest rats from group D1 and D2 were excluded. The food intake and weight were weighted daily and weekly,respectively. The subcutaneous,visceral and total fat contents of rats was measured by 256-row CT scan and the Lee index was calculated accordingly. The kidney,liver,testis,spleen and heart were weighted respectively. Serum leptin and insulin levels were quantified. The pathology in liver and adipose tissues were analyzed by HE staining. Oral glucose tolerance test(OGTT)was used to compare the glucose tolerance ability. Serum total cholestero(lT-CHO),high density lipoprotein(HDL-C),low density lipoprotein(LDL-C),triglyceride(TG)and inflammatory cytokines IL-6,TNF-α were detected as well. Results: After 12 weeks,the body weight,subcutaneous fat,visceral fat,total fat mass,wet weight of liver,kidney and heart,area under blood glucose curve and the levels of serum insulin,leptin,T-CHO,LDL-C,TG,IL-6 and TNF-α in group D2 were significantly increased compared to those of group C and group D1. HDL-C of group D2 was markedly lower than that in group C(P<0. 05). The visceral fat,total fat content and HDL-C in group D1 were significantly different from those of group C(P<0. 05). Steatosis and enlarged adipocyte were found in the livers of rats in group D1 and D2,and the lesions were more significant in group D2. Conclusion: Purified HF diet was more effective in inducing metabolic abnormalities,steatosis,peripheral chronic inflammation in obese SD rat models. But lard oil HF diet was more economical when only inducing visceral steatosis was required.

Differences in action potential propagation speed and axon initial segment plasticity between neurons from Sprague-Dawley rats and C57BL/6 mice

Action potentials(APs)in neurons are generated at the axon initial segment(AIS).AP dynamics,including initiation and propagation,are intimately associated with neuronal excitability and neurotransmitter release kinetics.Most learning and memory studies at the single-neuron level have relied on the use of animal models,most notably rodents.Here,we studied AP initiation and propagation in cultured hippocampal neurons from Sprague-Dawley(SD)rats and C57BL/6(C57)mice with genetically encoded voltage indicator(GEVI)-based voltage imaging.Our data showed that APs traveled bidirectionally in neurons from both species;forward-propagating APs(fpAPs)had a different speed than backpropagating APs(bpAPs).Additionally,we observed distinct AP propagation characteristics in AISs emerging from the somatic envelope compared to those originating from dendrites.Compared with rat neurons,mouse neurons exhibited higher bpAP speed and lower fpAP speed,more distally located ankyrin G(AnkG)in AISs,and longer Nav1.2 lengths in AISs.Moreover,during AIS plasticity,AnkG and Nav1.2 showed distal shifts in location and shorter lengths of labeled AISs in rat neurons;in mouse neurons,however,they showed a longer AnkG-labeled length and more distal Nav1.2 location.Our findings suggest that hippocampal neurons in SD rats and C57 mice may have different AP propagation speeds,different AnkG and Nav1.2 patterns in the AIS,and different AIS plasticity properties,indicating that comparisons between these species must be carefully considered.

不同低氧胁迫方式下SD大鼠急进高原模型的比较研究

目的对SD大鼠在高原实地和模拟高原环境这2种低氧胁迫方式下建立的急进高原模型进行比较研究,进而鉴定模拟高原实验舱的可靠性。方法将SD大鼠分别急进模拟高原动物实验舱(4000 m)或高原实地实验室(4010 m)来建立大鼠急进高原模型,在暴露24 h或72 h后采集并测定高原生理病理变化相关指标,主要包括血常规、血生化、血气、氧化损伤指标(超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px))、炎症指标(白细胞介素-1β(interleukin 1β,IL-1β)、γ干扰素(interferon-γ,IFN-γ)、单核细胞趋化蛋白1(monocyte chemotactic protein 1,MCP-1)和白细胞介素-6(interleukin 6,IL-6))、病理组织分析和低氧敏感基因(低氧诱导因子-1α(hypoxia inducible factor-1α,Hif-1α)和血管内皮生长因子A(vascular endothelial growth factor A,Vegfa))等,最终对结果进行差异性分析,得出差异性评估报告。结果相同海拔高度下,高原实地或模拟高原暴露72 h后均可以产生明显的肺部和脑部损伤。相同暴露时间下,动物机体的血常规、血生化和血气结果相近,炎症指标(IL-6,IL-1β,MCP-1和IFN-γ)、氧化损伤指标(MDA,SOD和GSH)和脑部低氧敏感基因(Hif-1α和Vegfa)等检测结果无显著性差异。但是,模拟72 h组的二氧化碳分压(partial pressure of carbon dioxide,PaCO_(2))和碱剩余(base excess,BE)显著高于其他低氧处理组、模拟72 h组的肺部低氧敏感基因(Hif-1α和Vegfa)与空白对照组无显著性差异,以及高原实地处理组脑系数显著高于模拟高原处理组等结果提示高原实地和模拟高原环境可能存在细微区别。结论模拟高原实验舱在4000 m海拔可成功建立急进高原动物模型,最优暴露时间应大于24 h但略短于72 h。模拟高原实验舱具有良好的可靠性,但条件允许情况下应尽量前往高原实地来建立急进高原动物模型。

Analysis of Behaviour Pattern in Sprague-Dawley Rats Using Microwave Techniques

This communication presents a new method of detecting the behaviour pattern in Sprague-Dawley (SD) rats based on the measurement of the dielectric properties of blood plasma at microwave frequencies at different periods of time. The microwave measurements were performed by rectangular cavity perturbation method in the S-band of microwave frequency with the blood plasma collected from normal rats (Controls) as well as chemically induced rats (A&beta;). A change is observed in the dielectric properties of the A&beta;samples but not the controls samples at the extended period of time. This measurement technique is simple and the collection of blood from the rats is nonsurgical in nature. These results prove a new method of diagnosing Alzheimer’s Disease (AD) using microwave techniques.

N-亚硝基布美他尼SD大鼠体内致突变性风险研究

目的评价布美他尼杂质N-亚硝基布美他尼的体内致突变风险和肝细胞DNA损伤风险。方法SD大鼠随机分为6组,分别为:溶媒对照组(溶媒为0.5%羧甲基纤维素钠),N-亚硝胺布美他尼100、300、1000 mg·kg^(-1)剂量组,阳性对照组1[N-乙基-N-亚硝基脲(ENU),40 mg·kg^(-1)]、阳性对照组2[甲磺酸乙酯(EMS),200 mg·kg^(-1)]。2个阳性对照组均为每组6只动物,其余每组12只动物。大鼠连续14 d经口灌胃给予受试物N-亚硝胺布美他尼。首次给药后约14 d和约28 d后采集外周血用于Pig-a基因突变率检测,末次给药后约3 h取肝细胞开展彗星试验。结果试验期间给予N-亚硝基布美他尼未导致异常临床症状和动物体重及摄食量改变。100、300、1000 mg·kg^(-1)剂量组动物肝细胞平均tail%DNA值(平均数/中位数)分别为2.90±0.38/2.34±0.46、3.58±0.27/2.87±0.51、3.45±0.59/2.21±1.44,与溶媒对照组相应数值相比未见差异,且无明显剂量效应相关性。首次给药后14 d,100、300、1000 mg·kg^(-1)剂量组动物平均RBC^(CD59-)和RET^(CD59-)百万分之发生率(RBC^(CD59-)百万分之发生率/RET^(CD59-)百万分之发生率)分别为2.9±1.6/1.2±0.8、2.8±2.4/1.3±1.5、2.4±1.0/1.3±1.5;首次给药后28 d,100、300、1000 mg·kg^(-1)剂量组动物RBC^(CD59-)百万分之发生率/RET^(CD59-)百万分之发生率分别为5.1±1.5/2.2±0.6、4.3±1.5/3.5±3.6、4.8±2.4/2.5±2.7。上述数值与相同时间点溶媒对照组相比均未见差异,且经统计学分析未见剂量效应相关性。结论连续经口给予N-亚硝基布美他尼14 d,SD大鼠的最大耐受量高于1000 mg·kg^(-1),未检出外周血基因突变风险和肝细胞DNA损伤风险。研究数据可为药品中遗传毒性杂质的合理监管和含N-亚硝基杂质药物的安全使用提供数据支持。

SD大鼠、小鼠创面新生皮肤组织石蜡切片的制备方法

目的:探索有效的SD大鼠、小鼠创面新生皮肤组织石蜡切片的制备方法。方法:取21只雄性SD大鼠、21只雄性SD小鼠制作创伤模型,在不同愈合时间取创面新生皮肤组织制作石蜡切片,对石蜡切片制作过程中的关键步骤进行控制,包括固定、脱水、浸蜡、包埋、切片、脱蜡、复水、染色、脱水、封藏等。比较不同愈合时间皮肤组织石蜡切片的差异。结果:实验制得组织切片染色均匀,细胞排列紧密,无裂缝,细胞核染色清晰,呈暗红色,成纤维细胞、毛囊、毛细血管和肌肉组织结构均清晰可见。随着愈合时间的延长,皮肤表皮层增厚,毛囊越来越多且分化程度提高,成纤维细胞数量增加。结论:该制备方法可以有效保证石蜡切片的质量并判定皮肤愈合程度。

不同抗凝比例对SD大鼠凝血指标的影响

目的探讨不同抗凝比例对SD大鼠凝血结果的影响。方法SD大鼠80只,雌雄各半。腹腔静脉采血,部分进行血钙(Ca^(2+))、血细胞比容(HCT)和血小板(PLT)的检测,其余按照枸橼酸钠抗凝管1∶2、1∶4、1∶6、1∶8和1∶10的抗凝比例,进行凝血酶原时间(PT)和活化部分凝血活酶时间(APTT)的检测,并分析结果。结果SD大鼠的Ca2+和HCT与人类正常值相近,但PLT远大于人类PLT的正常值范围。APTT随着抗凝剂量增加时间延长,各抗凝组间差异有统计学意义(P<0.05),PT随着抗凝剂量增加时间延长,各抗凝组与1∶2或1∶4组间差异有统计学意义(P<0.05)。APTT在抗凝比1∶8~1∶10时凝固曲线基线期缩短,PT在抗凝比1∶2~1∶4时凝固曲线有异常波动。结论SD大鼠最优抗凝比应适当高于人类,在1∶6左右能降低不合格标本的检出率,给动物实验提供更可靠的检测结果。

不同周龄和性别的SPF级SD大鼠血液生理生化指标的测定与比较分析

目的探讨分析不同周龄和性别的SPF级SD大鼠血液生理生化指标。方法研究对象为SPF级SD大鼠80只,并随机将其分为4、12、24、48周龄4个实验组,每组内雌雄比例为1:1,所有SD大鼠均应用XFA6030型动物血液细胞分析仪对其血液生理指标展开分析,应用Beckman Coulter AU5800全自动生化分析仪对其血液生化指标展开分析。结果与12、24、48周龄SD大鼠相比较,4周龄的SD大鼠在WBC、RBC、HGB、HCT、MCV、MCH、MCHC、PLT、PCT、BA#、BA%几方面具有较大差异,P<0.05,且各项指标达到峰值的时间均为24周,在雌雄的性别差异中,4周龄SD大鼠BA%具有差异;12、24周龄SD大鼠WBC、RBC具有差异;24周龄SD大鼠BA%具有差异;48周龄SD大鼠MCH、PCT具有差异,且所有差异P值均<0.05;与12、24、48周龄SD大鼠相比较,4周龄的SD大鼠在AST、CHOL、TP、GLU、CRE、ALT、ALP、BUN、TG几方面具有较大差异,P<0.05,在雌雄的性别差异中,4周龄SD大鼠CRE、ALP见具有差异;12周龄SD大鼠AST、CHOL、BUN具有差异;12周龄以及24周龄SD大鼠ALP具有差异,且所有差异P值均<0.05。结论在性别相同但周龄不同的SD大鼠中,其血液生理生化指标存在统计学差异,同样的,在同周龄但不同性别的SD大鼠中亦存在差异。

The effect of Xin-Nao-Xue-Mai-Ning on SD rats with hypothyroidism and IRE1-XBP1 signaling

Objective:To investigate the effect of Xin-Nao-Xue-Mai-Ning(XNXMN)on hypothyroidism in SD rats and explore the effect of IRE1-XBP1 signaling pathway on hypothyroidism in rats,and to analyze the possible mechanism of XNXMN in the treatment of hypothyroidism.Method:Twenty-four male SD rats were given a 0.1%concentration of propylthiouracil(PTU)solution by gavage every day to establish a hypothyroidism animal model.After the completion of the model building,intervention with XNXMN.Observe the general conditions of rats,serum triiodothyronine(T3),tetraiodothyronine(T4),thyroid stimulating hormone(TSH)levels,X box binding protein 1(XBP-1)protein expression level,and perform statistical analysis.Results:XNXMN can improve the general condition of hypothyroidism rats,reduce serum TSH,increase T3,T4 and reduce the expression of XBP1 protein in thyroid endoplasmic reticulum.Conclusion:XNXMN can improve the symptoms of hypothyroidism rats and has the effect of treating hypothyroidism.Its mechanism of action may be related to the expression of IRE1-XBP1 signaling pathway proteins that regulate the unfolded protein pathway of the endoplasmic reticulum.