Effects of Tongmai Huoxue Yin(通脉活血饮) on Tumor Necrosis Factor-alpha in the Acute Cerebral Ischemia Model Rat

Objective:To observe the interfering action of Tongmai Huoxue Yin(通脉活血饮) on the acute cerebral ischemia model rat.Methods:Total 60 SD rats,30 females and 30 males,were randomly divided into 4 groups,sham-operation group,model group,Nimodipine group and Tongmai Huoxue Yin group,15 rats in each group.The acute cerebral ischemia rat model was duplicated,the middle cerebral artery(MCA) were ligated and the thread was inserted for the rats in the model group,Nimodipine group and Tongmai Huoxue Yin group,for the rats in the sham-operation group,the arteries were separated without ligature and the thread was not inserted.After the modeling has succeed,the water-decocted concentrated solution of 20-fold Tongmai Huoxue Yin clinical dosage was intragastrically administrated in a dose of 3 mL /100 g · d divided into twice,1.5 mL /100 g once.Distilled water 3 mL /100 g·d was intragastrically administrated,1.5 mL /100 g once,for the rat in the model group,Nimodipne suspension 3 mL /100 g·d(0.6 mg /100 g) for the Nimodipine group and 3 mL /100 g · d(5.4g /100 g) for the Tongmai Huoxue Yin group,no drugs for the sham-operation group.And changes of tumor necrosis factor-alpha(TNF-α) contents in the serum and brain tissue were investigated.Results:Compared with the model group,compared with the sham-operation group,serum TNF-α content at 5 h of focal cerebral ischemic ischemia in the model group started to increase and reached to the high peak at 12 h,but in both the Tongmai Huoxue Yin group and the Nimodipine group decreased in varying degrees at the same time;compared with the sham-operation group,brain TNF-α content at 6 h of focal cerebral ischemic ischemia in the model group started to increase and reached to the high peak at 12 h,but in both the Tongmai Huoxue Yin group and the Nimodipine group decreased in varying degrees,with the most obviously decreased at 24 h of ischemia.Tongmai Huoxue Yin could significantly decrease TNF-α content in the brain tissue.Conclusion:Tongmai Huoxue Yin has a protective action on acute cerebral ischemia injury in the rat.

Nonhuman primate models of focal cerebral ischemia

Rodents have been widely used in the production of cerebral ischemia models. However, successful therapies have been proven on experimental rodent stroke model, and they have often failed to be effective when tested clinically. Therefore, nonhuman primates were recommended as the ideal alternatives, owing to their similarities with the human cerebrovascular system, brain metabolism, grey to white matter ratio and even their rich behavioral repertoire. The present review is a thorough summary of ten methods that establish nonhuman primate models of focal cerebral ischemia; electrocoagulation, endothelin-1-induced occlusion, microvascular clip occlusion, autologous blood clot embolization, balloon inflation, microcatheter embolization, coil embolization, surgical suture embolization, suture, and photochemical induction methods. This review addresses the advantages and disadvantages of each method, as well as precautions for each model, compared nonhuman primates with rodents, different species of nonhuman primates and different modeling methods. Finally it discusses various factors that need to be considered when modelling and the method of evaluation after modelling. These are critical for understanding their respective strengths and weaknesses and underlie the selection of the optimum model.

Effects of strain, body mass, and thread tip preparation on the establishment of focal cerebral ischemia mouse models

BACKGROUND： What are the successful factors of the establishment of the thread-blocking method for focal cerebral ischemia/reperfusion mouse models？ OBJECTIVE： To study the effects of strain, body mass, and thread tip preparation for the establishment of focal cerebral ischemia mouse models by using middle cerebral artery occlusion （MCAO）. DESIGN： Observational contrast animal study. SETTING： Gulou Hospital, Medical College of Nanjing University. MATERIALS： （1） The following experiment was performed at the Animal Experimental Center, Gulou Hospital Affiliated to Medical College of Nanjing University from December 2006 to April 2007. Sixty male white Kunming mice, whose body masses were 18-22 g （n =40）, 25-29 g （n =10） and 30-33 g （n =10）, as well as 10 male C57BL/6J mice, whose body mass was 18-22 g, were provided by the Animal Experimental Center, Gulou Hospital Affiliated to Medical College of Nanjing University. All mice were 10-12 weeks old. The project received confirmed consent from the local ethics committee. （2）Experimental materials： tripheryltetrazolium hydrochloride （TTC） and 0.1% poly-L-lysine were provided by Sigma Company, USA; citromint was provided by Shanghai Lingfeng Chemical Company Limited. METHODS： （1） Strain comparison： Ten white Kunming mice （weighing 18-22 g） and ten C57BL/6J mice （weighing 18 - 22 g） were selected. （2） Comparison of body mass： Thirty white Kunming mice, whose body masses were 18-22 g （n =10）, 25-30 g （n =10）, and 30-35 g （n =10）, were divided into groups. （3） Comparison of thread tip preparation： White Kunming mice weighing 18-22 g were divided into a poly-L-lysine line group and general line group, with 10 mice in each group. Mice in these two groups, which were respectively treated with poly-L-lysine or nothing, underwent MCAO. （4） All experimental mice received MCAO. Three hours after ischemia, and 24 hours after reperfusion, neurological deficit scores were measured and a success rate of model establishment was calculated. In addition, after sacrifice, sample tissues were cut into coronal sections to calculate the cerebral infarction area. MAIN OUTCOME MEASURES： （1） Success rate of model establishment and cerebral infarction area. （2） Neurological deficit scores. RESULTS： Sixty white Kunming mice and 10 C57BL/6J mice were included in the final analysis. （1） Strain comparison： The cerebral infarction area of white Kunming mice was larger than the C57BL/6J mice, and the neurological deficit scores of white Kunming mice were greater than those of the C57BL/6J mice （P 〈 0.05）. （2） Comparison of body mass： A success rate of model establishment in white Kunming mice weighing 18-22 g was higher than in white Kunming mice weighing 25-30 g or 30-35 g （P 〈 0.05）. （3） Comparison of thread tip preparation： The cerebral infarction area of mice in the poly-L-lysine line group was larger than in mice in the general line group. The neurological deficit scores of mice in the poly-L-lysine line group were greater than in mice in the general line group （P 〈 0.05）. CONCLUSION： Strain, body mass, and thread tip preparation can affect the establishment success rate of a focal cerebral ischemia mouse model using MCAO.

Effects of Chuanxiongqin hydrochloride on increasing the fluidity of brain cell membrane and scavenging free radicals in model rats with ischemia/reperfusion injury

BACKGROUND: The fluidity of cell membrane can be affected by various factors. Many experiments have confirmed that the ischemia/reperfusion of organic tissue can increase the contents of free radicals, which lead to high rigidity and low fluidity of cell membrane, and the conditions can be changed by Chuanxiongqin. OBJECTIVE: To observe the effect and mechanism of Chuanxiongqin hydrochloride on the fluidity of brain cell membrane in rat models of ischemia/reperfusion. DESIGN: A completely randomized controlled animal trial. SETTINGS: Institute of Brain Sciences; Department of Physiology, Medical College, Datong University. MATERIALS: Twenty male grade Ⅰ Wistar rats of 170-220 g were randomly divided into model group (n =10) and control group (n =10). Chuanxiongqin hydrochloride (molecular mass was 172.2) was purchased from the National Institute for the Control of Pharmaceutical and Biological Products (batch number: 0817-9803); Spin labelers: 5-doxyl-stearlic acid methylester (5DS), 16-doxyl-stearlic acid methylester (16DS), xanthine, xanthine oxidase (XOD) and 5,5-dimeth-1-pyrroline- N-oxide (DMPO) from Sigma Company; Bruker ESP 300 electron paramagnetic resonance (EPR) spectrometer by Bruker Company (Germany). METHODS: The experiments were carried out in the State Key Laboratory of Natural and Biomimetic Drugs, Peking University from June 2001 to July 2002. In the model group, rats were made into models of cerebral ischemia by 30-minute ligation and 2-hour reperfusion of common carotid arteries; The rats in the control group were not made into models. The order parameter (S) and rotational correlation time (τc) were detected with the ESR spectrometer by means of spin labeling. The greater the S and τc, the smaller the fluidity. Meanwhile, the clearance rate of free radicals was detected with ESR spin trapping. The measurement data were compared using the t test. MAIN OUTCOME MEASURES: The S, τc and clearance rates of O2 · and OH· free radicals were compared between the model group and control group. RESULTS: The S and τc in the model group [0.738 4±0.003 5; (8.472±0.027)×10-10 s/circle] were obviously different from those in the control group [0.683 9±0.008 3; (7.945±0.082)×10-10 s/circle, t =5.731, 5.918, P < 0.05], which suggested that ischemia/reperfusion injury decreased the fluidity of brain cell membrane. After adding Chuanxiongqin hydrochloride, there were no obvious differences between the model group [0.688 5±0.030 5; (7.886±0.341)×10-10 s/circle] and control group (P > 0.05), indicating that Chuanxiongqin hydrochloride could recover the fluidity of brain cell membrane after ischemia/reperfusion injury close to the level in the normal control group. Chuanxiongqin hydrochloride could directly scavenge the O2 · and OH· free radicals, and the maximal clearance rates were 83.92% and 44.99% respectively. CONCLUSION: Chuanxiongqin hydrochloride increases the fluidity of membrane of ischemia-injured brain cell by scavenging both O2 ·and OH· free radicals.

The rat model of placental ischemia as a model of postpartum posterior cortical atrophy?

Potential link between preeclampsia（PE）and posterior cortical atrophy（PCA）：PCA is a neurodegenerative disorder affecting the parietal,occipital,and occipital-temporal brain regions,often manifesting as a decline in visual processing and perception skills in affected individuals.

Meta-analysis of Buyang Huanwu decoction in animal model of cerebral ischemia

Objective:To evaluate the clinical efficacy of Buyang Huanwu decoction in treating animal model of cerebral ischemia.Methods:We searched PubMed,EMbase,Cochrane,CNKI,VIP,WanFang(1983 to 2021)for randomized control trials about Buyang Huanwu decoction treating animal model of cerebral ischemia.Results:According to the inclusion and exclusion criteria and assessment the quality of included trials(RCTs),the extraction of effective data,7 randomized clinical trials and their efficiency were evaluated,statistical analysis was conducted by using RevMan 5.3 software.The outcome measures assessed were neurological score and/or infarction volume.Meta-analysis showed that the neurological behavior scores of Buyang Huanwu decoction on animal model of cerebral ischemia was lower than the control group(SMD=2.06,95%CI(1.75,2.37),P<0.00001);The cerebral infarct volume of Buyang Huanwu decoction on animal model of cerebral ischemia were smaller than the control group(SMD=4.08,95%CI(3.57,4.58),P<0.00001).Conclusion:It was effective by using Buyang Huanwu decoction to reduce neurological behavior scores and the volume of animal model of cerebral ischemia.

Gradual Clamping Reduced Ischemia-Reperfusion Injury in an Isolated Rat Heart Model

Objectives: We hypothesized that the organisms and their organs or tissues could adapt themselves to the gradual changes of environment for surviving or reducing damage. This study explored whether gradual clamping (GC) could reduce myocardial ischemia-reperfusion (IR) injury in rat heart. Methods: Twelve rats were randomized to IR group and GC group, then the hearts were isolated and perfused with Langendorff apparatus. Before cardioplegia, the perfusion was stopped abruptly in IR group while slowly with 5-minute in GC group. The hearts were subjected to 30-minute ischemia and 60-minute reperfusion. The left ventricular develop pressure (LVDP) and systolic pressure (LVSP), the maximal rate of the increase and decrease of left ventricular pressure (+dp/dtmax, ﹣dp/dtmax) were measured by polygraph system at different time points. The recovery of the variables was expressed as the ratio of these values at individual time point after reperfusion to the baseline respectively. Results: The recovery of LVDP after reperfusion was better than that in IR group (P = 0.034). No significant difference in the recovery of LVSP, +dp/dtmax and ﹣dp/dtmax between groups was observed. Conclusions: Gradual clamping could improve the recovery of LVDP after IR, suggesting that gradual clamping could reduce myocardial IR injury.

内皮细胞特异性骨形态发生蛋白2对血管新生的影响:生物信息学分析和实验验证

背景:血管新生是心血管疾病的主要干预靶点,骨形态发生蛋白2具有调控血管新生作用,但内皮细胞特异性骨形态发生蛋白2对血管新生的调控作用不清楚。目的:探讨内皮细胞特异性骨形态发生蛋白2对血管新生的影响。方法:(1)生物信息学分析:通过Panglao DB公共基因表达数据库单细胞转录组荟萃分析观察骨形态发生蛋白2细胞群表达丰度和定位。血管新生小鼠和内皮(心内膜)过表达骨形态发生蛋白2小鼠转录组测序数据集探索内皮细胞骨形态发生蛋白2对血管新生信号通路的调控作用。(2)体内实验验证:建立小鼠后肢缺血模型,对比模型小鼠患侧与健侧缺血后肢7,14和21 d血流灌注情况,免疫荧光和免疫组织化学染色评估小鼠骨形态发生蛋白2和CD31的表达定位情况。(3)体外实验验证:体外培养人脐静脉内皮细胞,分为对照组、缺氧组和骨形态发生蛋白2抑制剂(Noggin蛋白)干预组,培养24 h,观察各组内皮细胞血管新生情况。结果与结论:(1)内皮细胞是表达骨形态发生蛋白2的重要细胞亚群,在血管新生内皮细胞和骨形态发生蛋白2过表达内皮细胞转录组再分析均发现骨形态发生蛋白2表达明显升高,血管新生通路明显激活。(2)缺血7 d小鼠新生血管周围骨形态发生蛋白2阳性血管明显增加(P<0.05),缺血2周骨形态发生蛋白2阳性血管明显减少(P<0.001)。(3)体外培养人脐静脉内皮细胞,缺氧干预后,内皮细胞迁移能力和血管出芽明显增加,血管新生因子血管内皮生长因子和血小板衍生生长因子的表达明显升高,Noggin明显减少了缺氧诱导的内皮细胞血管新生(P<0.001),并下调血管内皮生长因子和血小板衍生生长因子的表达(P<0.01)。(4)结果证实,内皮细胞特异性骨形态发生蛋白2具有调控血管新生作用,靶向性内皮细胞骨形态发生蛋白2可望改善血管新生。

基于灰阶超声构建症状性颈动脉斑块影像组学模型的临床研究

目的:探讨基于灰阶超声构建症状性颈动脉斑块影像组学模型的可行性及应用价值。方法:由两名超声医师独立收集颈动脉斑块病例分别作为训练集和验证集。在训练集和验证集内部根据患者近期是否出现对应的脑缺血症状分为症状组和非症状组。比较两组患者一般临床资料情况。提取基于灰阶超声图像的颈动脉斑块影像组学特征。在训练集中,以是否存在脑缺血症状为因变量,以斑块影像组学特征为自变量,采用LASSO回归筛选自变量构建影像组学模型。采用ROC曲线、校准曲线及决策曲线对模型进行内部验证和外部验证。结果:训练集和验证集内部症状组和非症状组一般临床资料比较差异无统计学意义(P均>0.05)。训练集中共精选出6个斑块影像组学特征构建模型,该模型在训练集中的AUC为0.913[95%CI(0.862~0.964)],C-index为0.887;在验证集中的AUC为0.837[95%CI(0.758~0.917)],C-index为0.798。训练集和验证集中校准曲线与理想曲线拟合良好,DCA曲线净获益值高。结论:基于灰阶超声构建影像组学模型识别症状性颈动脉斑块具有可行性及一定的应用价值。

缺血缺氧脑瘫大鼠的时效研究

目的:基于缺血缺氧脑瘫大鼠神经功能评分(Zea-Longa评分)、脑组织肉眼观和大脑海马区胱天蛋白酶-9(Caspase-9)、胱天蛋白酶-3(Caspase-3)的表达水平变化,探讨缺血缺氧模型脑瘫大鼠的有效时长。方法:选取3周龄斯泼累格·多雷(SD)健康大鼠,随机分为正常组和模型组,采用改良的Rice-Vannucci方法建立脑瘫模型,造模后第1、7、14、21天,观察各组大鼠的一般情况并进行神经功能评分,在第7、14、21天分批处死大鼠并取脑组织,观察各组大鼠左侧脑组织,检测海马区Caspase-9、Caspase-3的表达水平。结果:一般情况:造模后第1天,与正常组比较,模型组大鼠左侧瞳孔缩小、姿势异常、自发或夹尾左旋、自主活动减少、兴奋性降低、肌肉颤动、头颤,抽搐,抓取时抵抗反应明显,随着时间延长,以上异常行为逐渐消失,造模后21 d基本消失不见,但左侧瞳孔一直小于对侧;Zea-Longa评分:与正常组比较,模型组造模后7、14 d Zea-Longa评分较高,差异有统计学意义(P<0.05);脑组织肉眼观:与正常组比较,模型组造模后7、14及21 d大鼠左侧脑组织有不同程度的萎缩和坏死;免疫组化结果:与正常组比较,模型组造模后7 d、14 d Caspase-9、Caspase-3的表达水平均显著升高,差异有统计学意义(P<0.05)。结论:3周龄缺血缺氧脑瘫模型大鼠的有效时长为14~21 d,可干预14 d。

长爪沙鼠脑缺血和听觉障碍模型研究进展

目前分布在世界各地的实验动物长爪沙鼠均源自中国。早在1930年代,野生长爪沙鼠经人工驯养后被引入医学研究。时至今日,长爪沙鼠已经成为一种公认的“多功能实验动物”,被广泛应用于脑神经、寄生虫和微生物、肿瘤等研究领域。长爪沙鼠具有独特的脑底动脉解剖学特点,例如先天性Willis环缺失。因此,用单侧颈总动脉结扎的简易手术操作既可以构建脑缺血或脑缺血再灌注损伤模型,还可以实现同体对照。长爪沙鼠的这一解剖特点不仅增加了脑缺血的敏感性,也易于诱导耳蜗缺血。因此,长爪沙鼠在制备听觉障碍模型中也发挥重要作用。长爪沙鼠脑缺血模型和听觉障碍模型的发病过程和病理表现均与人类患者有诸多相似之处。科学家们利用长爪沙鼠构建了脑缺血模型、脑缺血再灌注损伤模型、耳蜗缺血模型、人工耳蜗植入模型、感音性神经性耳聋模型等,均取得显著成效。本文重点阐述当前长爪沙鼠脑缺血模型和听觉障碍模型的创制方法和评价指标,讨论各种造模方法的优缺点及其应用进展,以期为长爪沙鼠在这两个重要领域的应用提供理论依据和借鉴。

小鼠肠缺血再灌注损伤模型的构建与应用

肠缺血再灌注损伤(IRI)是临床上常见的急危重症,发病率和死亡率均较高.目前对肠IRI的基础研究得到广泛关注,而成功地构建肠IRI动物模型是开展基础研究的基础,夹闭肠系膜上动脉的肠IRI动物模型应用最为广泛.本文详细阐述了夹闭肠系膜上动脉的小鼠肠IRI模型的构建过程及注意事项,将有助于提高小鼠肠IRI模型构建的成功率,从而助推肠IRI的动物体内研究.

缺血性卒中无复流动物模型建立和多维度评价方案

目的 建立脑无复流模型并对灌注减低进行多维度评价。方法 利用激光散斑对比成像和双光子活体成像,比较C57BL/6(n=16)和BALB/c小鼠(n=37)短暂性大脑中动脉闭塞,以及BALB/c小鼠缺血1h或1.5h灌注改变情况。结合激光散斑对比成像、低倍率和较高放大倍率的灌注脑片图像,以及双光子显微镜监测红细胞流速和流量对短暂性大脑中动脉闭塞后脑灌注下降进行活体动态以及全脑切片和微血管的评估。用微管相关蛋白2染色和行为学评分评估梗死面积和行为学缺损。结果 在C57BL/6小鼠中,大脑中动脉区域大多数毛细血管在缺血期间仍然流动,而在BALB/c小鼠中,大多数毛细血管被阻断。此外,BALB/c小鼠缺血1.5 h后,24 h后再通时的皮质灌注减少至基线76.1%(与BALB/c sham组89.0%相比减少,P=0.046),而缺血1 h减少至79.9%,与BALB/c sham组无明显差异(P=0.299)。切片全脑灌注评估BALB/c小鼠短暂性大脑中动脉闭塞1.5 h导致全脑灌注减少至75.1%(与BALB/c sham组100%相比降低,P<0.001),双光子活体成像评估大脑中动脉流域皮质表面毛细血管血流在再通24h,红细胞流速降至基线水平的50.3%(与BALB/c sham组110.7%相比降低,P=0.010),红细胞流量降至基线水平的38.9%(与BALB/c sham组119.2%相比降低,P=0.010);高倍率切片成像评估毛细血管有灌注长度为241μm±33μm(与BALB/c sham组997μm±103μm相比减少,P=0.001),有灌注的毛细血管占据的总体积分数为0.0128±0.0018(与BALB/c sham组0.0539±0.0055相比减少,P<0.001)。微管相关蛋白2染色提示BALB/c小鼠短暂性大脑中动脉闭塞1.5 h导致梗死面积占全脑面积约36%,行为学评分提示行为学缺损,评分升高至9分。结论BALB/c小鼠短暂性缺血1.5 h导致明显的脑灌注下降以及毛细血管无复流,适合建立无复流模型。激光散斑对比成像、低倍率和较高放大倍率的灌注脑片图像,以及双光子显微镜活体成像的结合,可以对灌注变化进行多维度评价。

阿托伐他汀预处理对高血糖诱导小鼠脑缺血后出血转化的影响

目的探究阿托伐他汀对高血糖诱导的小鼠脑缺血后出血转化(HT)的作用及机制。方法36只SPF级雄性C57BL/6小鼠随机分为假手术组、HT模型组和阿托伐他汀组,每组12只。比较各组小鼠神经功能评分、死亡率、HT发生率、HT分级评分,苏木精-伊红染色观察脑组织出血情况,免疫荧光染色评估血脑屏障通透性,Western blot检测缺血半暗带脑组织免疫球蛋白G(IgG)、闭锁连接蛋白1(ZO-1)、闭合蛋白(occludin)、紧密连接蛋白5(claudin5)、基质金属蛋白酶(MMP)2和MMP-9的蛋白表达。结果与假手术组比较,HT模型组神经功能评分、死亡率、HT发生率、HT评分、IgG荧光强度、IgG、MMP-2、MMP-9蛋白表达水平显著增高,ZO-1、occludin、claudin5蛋白表达水平明显降低(P<0.01)。与HT模型组比较,阿托伐他汀组神经功能评分、死亡率、HT发生率、HT评分、IgG荧光强度及IgG、MMP-2、MMP-9蛋白表达水平显著降低[(2.73±1.19)分vs(3.91±0.94)分,16.7%vs 41.6%,58.3%vs 91.6%,(1.00±1.04)分vs(2.58±1.13)分,(504.30±105.52)a.u vs(859.91±153.28)a.u,4.55±1.40 vs 12.06±3.73,1.87±0.41 vs 2.95±0.68,1.47±0.24 vs 2.12±0.23,P<0.05,P<0.01],ZO-1、occludin、claduin5蛋白表达显著升高(1.55±0.20 vs 0.53±0.10,0.92±0.11 vs 0.35±0.07、0.58±0.04 vs 0.30±0.05,P<0.01)。结论阿托伐他汀可通过抑制MMP-2、MMP-9激活,上调ZO-1、occludin、claudin5表达,降低血脑屏障通透性,从而抑制高血糖诱导的脑缺血后HT。

张家口地区急性心肌梗死患者行急诊PCI时发生缺血再灌注损伤的预测模型

目的 构建经皮冠状动脉介入(percutaneous coronary intervention, PCI)术后发生缺血/再灌注损伤的预测模型,并评价其有效性。方法 收集张家口地区262例急性心肌梗死并行急诊PCI患者的临床资料,根据PCI术后是否发生缺血/再灌注损伤分为缺血/再灌注损伤组和非缺血/再灌注损伤组,应用多因素Logistic回归筛选独立危险因素,绘制列线图对缺血/再灌注损伤发生率进行预测。利用ROC下面积(AUC),校准曲线和DAC曲线评价其鉴别力、校准能力和临床应用价值。使用Hosmer-Lemeshow检验评估校准度,使用内部抽样法进行验证。结果 构建不同模型取最小AIC值,最终将收缩压、Killip分级、多支病变、BNP升高、白介素6、超氧化物歧化酶纳入预测模型中绘制列线图,AUC面积为0.818(95%CI:0.748~0.888),校准曲线显示预测结果和实际结果有较好的一致性,决策曲线表明列线图具有临床实用价值。结论 缺血/再灌注损伤预测模型有较高的区分度和准确性,可以筛选高危人群,有较高的预测价值。

不同线栓法脑缺血模型对小鼠脑区及行为的影响

目的 探讨3种线栓法模型对小鼠不同脑区缺血及小鼠行为学差异的影响。方法 将120只ICR小鼠随机分成4组:假手术组、A组(颈总动脉进线栓至颈内动脉8 mm)、B组(颈外动脉进线栓至颈内动脉10 mm)、C组(颈总动脉进线栓至颈内动脉10 mm),每组30只。脑缺血1.5 h再灌注24 h后,采用2,3,5-氯化三苯基四氮唑(TTC)染色观察小鼠脑缺血部位并计算缺血面积,通过Longa评分评估小鼠神经运动功能,通过抓力实验评估小鼠的前肢力量,通过游泳实验评估小鼠的体力,伊文思蓝染色测定血脑屏障的通透性,通过计算脑含水量评估脑水肿程度,苏木精-伊红(HE)染色观察脑组织病理改变。结果 TTC染色结果显示3种方法造成小鼠大脑梗死的区域及面积有所不同,A、B组在梗死面积上差异无统计学意义(P>0.05),但C组梗死面积相较于A、B组差异均有统计学意义(P<0.05),假手术组未发现梗死区域。行为学实验结果显示A、B、C 3组结果相比于假手术组差异均有统计学意义(P<0.05),Longa评分中B、C组得分差异无统计学意义(P>0.05),但A组相较于B、C组差异均有统计学意义(P<0.05);前肢抓力实验中仅A、C组测试结果差异有统计学意义(P<0.05);游泳实验B、C组游泳时间差异无统计学意义(P>0.05),但A组相较于B、C组差异均有统计学意义(P<0.05)。伊文思蓝染色、脑含水量以及HE染色发现相比于A、B组,C组所造成的脑缺血对小鼠脑损伤更严重。结论 3种线栓法脑缺血模型造成小鼠大脑不同脑区的缺血,且小鼠皮质缺血更容易造成运动功能障碍。

精胺后处理上调线粒体自噬保护脑缺血/再灌注损伤小鼠模型的分子机制研究

目的 探讨精胺后处理保护脑缺血/再灌注损伤(I/RI)小鼠模型的疗效及其分子机制。方法 60只6~8 w龄C57BL小鼠随机分为6组(每组10只):(1)对照(Control)组;(2)假手术(Sham)组;(3)模型(MCAO)组;(4)模型+精胺治疗(MCAO+Spm)组;(5)MCAO+Spm+Adv shRNA-Drp-1组;(6)MCAO+Spm+Adv shRNA-NT组。构建大脑中动脉闭塞(MCAO)小鼠模型。TTC(2,3,5-氯化三苯基四氮唑)染色脑切片测定脑梗死体积。尼氏(Nissl)染色脑组织切片测定神经元死亡情况。ELISA测定活性氧自由基(ROS)、超氧化物歧化酶(SOD)和丙二醛(MDA)水平。腺病毒转染敲低脑组织动态相关蛋白-1 (Drp-1)。Western blot测定脑组织Drp-1、B淋巴细胞瘤基因-2相关X蛋白(Bax)、B淋巴细胞瘤基因-2(Bcl-2)、PTEN诱导激酶1(PINK1)和自噬相关蛋白(Parkin-1)的表达情况。结果 与Sham组相比,MCAO组脑梗死体积所占百分比升高(P<0.05),Nissl阳性细胞所占百分比降低(P<0.05),ROS和MDA水平升高(P<0.05),SOD水平降低(P<0.05),Drp-1、Bax、PINK1和Parkin-1的表达水平升高(P <0.05),Bcl-2表达水平降低(P<0.05)。与MCAO组相比,MCAO+Spm组脑梗死体积所占百分比降低(P<0.05),Nissl阳性细胞所占百分比增多(P <0.05),ROS和MDA水平降低(P <0.05),SOD水平升高(P <0.05),Drp-1、Bcl-2、PINK1和Parkin-1的表达水平升高(P<0.05),Bax的表达水平降低(P<0.05)。与MCAO+Spm组相比,MCAO+Spm+Adv shRNA-Drp-1组Drp-1和Bax表达水平降低,Bcl-2、PINK1和Parkin-1的表达水平升高(P <0.05)。结论 Spm后处理通过Drp-1上调线粒体自噬发挥保护I/RI损伤小鼠模型的功效。

多肽CRY-14保护心肌对抗缺血缺氧损伤的作用机制

目的研究多肽CRY-14在心肌缺血缺氧中的作用与机制。方法分析多肽CRY-14对缺血缺氧后H9C2心肌细胞增殖、氧化应激的影响。通过M型小动物心超比较多肽CRY-14对小鼠心肌梗死后心脏功能[左心室舒张末内径(LVEDD)、左心室收缩末内径(LVESD)、左心室射血分数(LVEF)及左心室缩短分数(LVFS)]的影响。小鼠心脏组织HE染色、Tunel染色以及Masson染色评估多肽CRY-14干预后小鼠心肌梗死后心脏组织学及凋亡的变化。此外,Western Blot实验初步探讨多肽CRY-14发挥心肌保护功能的机制。结果在细胞水平,多肽CRY-14可以缓解缺血缺氧对H9C2心肌细胞增殖的抑制作用,减轻缺血缺氧导致的氧化应激反应。在体水平,CRY-14可以缓解小鼠心肌梗死后心功能的改变,提高LVEF、LVFS,降低LVEDD、LVESD。CRY-14可以明显改善小鼠心肌梗死后心肌损伤,减轻心肌细胞凋亡以及心肌纤维化程度。此外,Western Blot结果提示CRY-14可以下调缺血缺氧后H9C2心肌细胞凋亡相关蛋白Caspase 3以及PARP的表达,CRY-14可以上调缺血缺氧后H9C2心肌细胞HSP70的表达。结论多肽CRY-14可通过改善缺血性心脏病中的氧化应激和凋亡水平发挥心肌保护作用,其机制可能与调控HSP70有关。

电凝法与线栓法制备大脑中动脉缺血模型在脑卒中后中枢痛研究中的应用对比

目的采用电凝法与线栓法分别造成小鼠大脑中动脉缺血,建立脑卒中后中枢痛(central post-stroke pain,CPSP)模型,探究更贴近临床的造模方法。方法选择6~8周龄(20~25g)健康雄性C57BL/6小鼠,随机分为空白对照组(Naive组)、电凝组(dMCAO组)和线栓组(tMCAO组),进行不同造模处理。在造模后行Longa神经功能缺陷评分,利用TTC染色评估大脑梗死体积,通过机械性缩足阈值和热缩足潜伏期评估小鼠的疼痛状态,旷场实验评估小鼠的运动功能。结果与Naive组相比,电凝组和线栓组小鼠造模后神经功能缺陷评分均升高(均P<0.01),TTC染色可观察到不同程度的脑缺血(P<0.05,P<0.01);与Naive组相比,电凝组和线栓组在造模后的第7、14、21、28天均表现出机械性痛觉超敏和热痛觉过敏,两组差异无统计学意义;与Naive组相比,电凝组小鼠在造模后第29天的运动功能无显著差异,而线栓组小鼠的运动功能下降(P<0.01)。结论电凝法和线栓法均可诱发脑卒中后中枢痛,但电凝法更贴近CPSP的临床表征,更具复制意义。

冠心病患者心肌缺血再灌注损伤的危险因素分析及预测模型效能研究

目的:分析冠心病患者心肌缺血再灌注损伤(MIRI)的危险因素,建立预测模型并评估效能。方法:选择2021年10月至2022年9月东莞市东南部中心医院接受溶栓治疗或经皮冠状动脉介入(PCI)治疗的冠心病患者80例为对象,根据患者治疗后是否发生MIRI分为对照组(未发生MIRI 45例)和观察组(发生MIRI 35例)。查阅所有患者的临床资料,对冠心病患者MIRI可能的影响因素进行统计,建立MIRI列线图预测模型,并评估模型预测效能。结果:80例冠心病患者中,有35例发生MIRI(43.75%)。多因素logistic回归结果显示,发作到手术时间、左室射血分数(LVEF)、中性粒细胞计数与淋巴细胞计数比值(NLR)、血小板计数与淋巴细胞计数比值(PLR)、肌钙蛋白T(TNT)、脑利钠肽(BNP)是冠心病患者MIRI发生的独立危险因素(P<0.05);本研究构建的列线图预测模型校准曲线斜率较高,霍斯莫-莱梅肖(H-L)拟合优度检验的结果为χ^(2)=1.334,P=0.323,表明模型拟合良好;受试者工作特征(ROC)曲线结果显示,曲线下面积为0.817,95%CI(0.759,0.871)。结论:发作到手术时间、LVEF、NLR、PLR、TNT及BNP是冠心病患者发生MIRI的独立危险因素,且基于上述因素构建的预测模型具有较高的拟合度,预测效能较高。