Effects of Total Flavonoids ofHippophae RhamnoidesL.on Intracellular Free Calciumin Cultured Vascular Smooth Muscle Cells of Spontaneously Hypertensive Rats and Wistar-Kyoto Rats

Essential hypertension (EH) has become oneof the main diseases that endanger human healthinmodern society . It has been a key project forscholars at home and abroad to find out howto ef-fectively prevent and cure hypertension andlesionsinits target organs . Total flavonoids ofHippophae rhamnoidesL.(TFH) ,a natural flavonoids ,is extractedfromthe fruit of wild plantHippophae rhamnoidesL.,the great mass of which is quercetin ( Que) andisorhamnetin (Isor) . Research has found thatTFH has di…     

大鼠脑缺血模型的制备方法与评价指标总结

本文综述了大鼠脑缺血模型的制备方法与评价指标总结，并提出了其优缺点，以供生物医药研究实验时根据需要的选择使用。     

Lewis大鼠骨髓间充质干细胞分离培养和鉴定

目的建立从Lewis大鼠骨髓中分离、培养间充质干细胞的方法,并对分离、培养的间充质干细胞进行鉴定.方法采用Percoll（1.073g/L）密度梯度分离和贴壁培养相结合的方法分离培养Lewis大鼠的骨髓间充质干细胞（MSC）.MSC细胞鉴定：采用相差显微镜观察MSC的形态学特征;流式细胞仪检测细胞表面标志抗原CD29,CD90,CD34和CD45的表达率;苏丹Ⅳ染色检测MSC成脂细胞分化;碱性磷酸酶染色和茜素红染色检测MSC成骨分化.结果原代培养的MSC于24 h后贴壁,48～72 h形成集落,12～15 d可达到80%～90%融合.第1代细胞表面标记物CD29,CD90,CD34和CD45的阳性率分别为80.41%,66.27%,2.73%,0.74%.第3代细胞表面标记物CD29,CD90,CD34和CD45的阳性率分别为89.91%,88.17%,0.81%,0.17%.细胞周期分析显示,第5代细胞G0/G1期细胞占68.9%,S＋G2＋M期为31.1%.用α-MEM培养液培养的第6代细胞部分分化为成骨和成脂细胞.结论采用Percoll（1.073g/L）密度梯度分离和贴壁培养相结合的方法能够成功分离和培养大鼠的MSC.第5代MSC细胞保持分化潜能.     

局灶性脑缺血后室管膜/室下区细胞迁移到梗塞区周围并分化为神经元和星形胶质细胞

目的研究局灶性脑缺血后室管膜/室下区细胞的迁移分化,揭示梗塞区周围新生细胞的来源.方法大脑中动脉阻塞前,将10μl 0.2%的荧光染料DiI注射于体质量250～350 g的雄性SD大鼠侧脑室以预标记室管膜/室下区细胞.脑缺血后,采用累积式的BrdU标记方法标记新生细胞并通过双重免疫荧光染色确定细胞分化.标记的细胞通过激光共聚焦显微镜观察.结果在非缺血对照大鼠,DiI标记细胞定居于室管膜/室下区.局灶性脑缺血后,DiI标记细胞出现于胼胝体,邻近的纹状体和皮质.此外,缺血14 d后,梗塞区周围纹状体和皮质内可见一些DiI/BrdU/GFAP或DiI/BrdU/NeuN三重标记阳性细胞.结论局灶性脑缺血后,室管膜/室下区细胞迁移到梗塞区周围并分化成神经元和星形胶质细胞,这一发现对于理解成体神经干细胞的起源和开发促进脑损伤后内源性神经发生的新措施具有重要意义.     

高压氧对缺氧缺血性脑损伤新生大鼠的远期影响

【目的】探讨早期和延迟高压氧（HBO）治疗对缺氧缺血性脑损伤（HIBD）新生大鼠的远期影响。【方法】7日龄大鼠37只随机分为对照组（n=10，假手术处理）、HIBD组（n=9）、EHBO组（n=9，HIBD模型后0．5～1h开始2个绝对大气压HBO治疗，稳压30min／次，间隔24h，连续2次）和LHBO组（n=9，HIBD模型后48～72h开始2个绝对大气压HBO治疗，稳压30min／次，间隔24h，连续5次），以大鼠远期的学习记忆功能（Morris水迷宫实验）和海马形态组织学（海马锥体细胞层结构和CAl区存活神经元数）来判断干预效果。【结果】HIBD组大鼠的学习记忆功能严重不良伴海马结构严重受损，与对照组相比，水迷宫实验中平均逃逸潜伏期延长（56．35S与23．07S，P〈0．05）、搜索时间和搜索路程缩短（分别为29．29S与51．21S，548cm与989cm．均P〈0．05）、海马存活神经元减少（100个／mm与183个／mm，P〈0．05）；早期HBO治疗明显减轻HIBD大鼠的学习记忆功能障碍（潜伏期：39．17s与56．35s；搜索时间：36．84s与29．29s；搜索路程：686cm与548cm，P〈0．05），并使海马存活神经元增多（131个／mm与100个／mm，P〈0．05），延迟HBO治疗不能减轻HIBD大鼠的学习记忆功能障碍（潜伏期：56．25s与56．35。；搜索时间：30．11s与29．29s；搜索路程：572cm与548cm，P〉0．05），海马存活神经元无增多（95个／mm与100个／mm，P〉0．05）；大鼠在水迷宫实验中的逃逸潜伏期与海马锥体细胞层存活神经元数呈直线负相关（r=-0．819，P〈0．01）。【结论】早期HBO治疗可减轻HIBD程度，并可改善HIBD所引起的远期学习记忆功能缺陷，延迟HBO治疗对HIBD无效。     

灵芝孢子油干预治疗6-羟多巴帕金森病大鼠模型的实验研究

[目的]研究灵芝孢子油对6-羟多巴(6-OHDA)帕金森病(PD)大鼠模型行为学及病理改变的影响,探讨该药对黑质多巴胺能神经元的保护作用.[方法]100只SD大鼠随机分为正常对照组20只、6-OHDA组40只、灵芝孢子油 6-OHDA组40只.通过脑部立体定向法将6-OHDA注射到SD大鼠一侧黑质致密部建立6-OHDA大鼠PD模型,灵芝孢子油 6-OHDA组在造模前3 d开始给灵芝孢子油每天500 mg/kg,连续10 d.6-OHDA组喂食生理盐水做对照.造模后每周给予阿朴吗啡观察大鼠旋转行为的变化,4周后用高效液相色谱法检测大鼠纹状体多巴胺及其代谢物含量,免疫组织化学法检测黑质致密部酪氨酸羟化酶(TH)阳性细胞数量,western blot法对TH蛋白进行半定量测定.[结果]①灵芝孢子油 6-OHDA组出现旋转行为的大鼠比例为10%,6-OHDA组为45%,两组比较有显著性差异.②纹状体多巴胺及其代谢物含量在两组间存在差异,多巴胺手术侧含量/手术对侧含量在灵芝孢子油 6-OHDA组为(60.12±7.5)%,在6-OHDA组为(38.58±7.26)%.③在黑质致密带区灵芝孢子油 6-OHDA组的TH免疫阳性细胞及TH蛋白表达量均较6-OHDA组显著增多.[结论]灵芝孢子油能明显改善6-OHDA大鼠模型行为学,增加纹状体多巴胺及其代谢物含量并提高黑质多巴胺能神经元的残存率,提示灵芝孢子油可能具有减缓PD病变进程的神经保护作用.     

大鼠佐剂性关节炎模型的建立与评价

目的诱导大鼠产生佐剂性关节炎（adjuvant arthritis，AA）并进行综合评价。方法大鼠左后足垫皮下注射完全弗氏佐剂（CFA）诱导AA后，采用关节炎评分法对关节肿胀程度进行评估，应用ELISA法检测大鼠血清中抗Ⅱ型胶原（Collagen Ⅱ,CⅡ）抗体的含量，并用透射电镜观察眉窝淋巴结和病变关节滑膜的超微结构的变化。结果注射CFA后模型组血清中抗CⅡ抗体含量明显升高，透射电镜显示呈典型的类风湿性关节炎病变。结论CFA能成功诱导大鼠AA模型，为研究类风湿性关节炎（rheumatoid arithritis，RA）的发病机制及药物疗效提供理想的实验模型。     

淫羊藿提取物对去卵巢大鼠骨转换率增强的抑制作用

目的研究淫羊藿提取物对去卵巢大鼠骨转换的影响。方法取3月龄SD大鼠手术切除双侧卵巢,1个月后ig给予阳性药17β-雌二醇(2mg/kg)以及淫羊藿提取物混悬液(110mg/kg),连续给药3个月。采用全自动生化分析仪测定血清中钙(s-Ca)、磷(s-P)的量和碱性磷酸酶(ALP)的活性及尿中钙(u-Ca)、磷(u-P)和肌酐(Cr)的量。利用ELISA试剂盒测定尿脱氧吡啶酚(DPD)的量。取胫骨近端制成不脱钙骨切片进行骨组织形态计量学分析。结果与假手术组相比,去卵巢大鼠s-Ca、s-P没有变化,ALP活性增强,u-Ca排出显著升高,尿中DPD与Cr的比值(DPD/Cr)显著增加,子宫萎缩。骨小梁面积百分比降低,骨小梁间隙增加。ig给予雌二醇或淫羊藿提取物3个月,血清ALP活性降低,u-Ca排出量减少,尿中DPD/Cr降低,骨小梁增加,且骨小梁间隙减小。雌二醇组大鼠子宫增生,淫羊藿组大鼠子宫与模型组相比没有变化。结论淫羊藿提取物能够显著抑制由于雌激素缺乏引发的骨转换增强,调整骨形成和骨吸收的关系,对遏制去卵巢造成的骨质疏松有一定的作用,且长期服用对子宫没有刺激作用。     

Dopamine-independent action of cocaine on striatal and accumbal neurons

Increasing evidence suggests that dopamine (DA) mechanisms alone cannot fully explain the psychoemotional and behavioural effects of cocaine, including its ability to induce drug-taking behaviour. Although it is known that cocaine, after intravenous administration or smoking, may reach brain levels high enough to inhibit Na+ transport, the role of this action remains unclear. To examine the contribution of local anaesthetic and DA mechanisms to changes in striatal and accumbal neuronal activity induced by cocaine, single-unit recording was combined with iontophoresis in awake, unrestrained rats. Most spontaneously active and glutamate-stimulated neurons were highly sensitive to brief cocaine applications (0-40 nA); cocaine-induced inhibitions occurred at small ejection currents (0-5 nA), were dose-dependent, highly stable during repeated applications and strongly dependent on basal activity rates. These neuronal responses remained almost unchanged after systemic administration of either a selective D1 antagonist (SCH-23390, 0.2 mg/kg) or a combination of SCH-23390 (1 mg/kg) and eticlopride (1 mg/kg), a D2 antagonist. Whereas SCH-23390 alone had a weak attenuating effect, no effect and even a slight enhancement of responses to cocaine occurred in fast-firing glutamate (GLU)-stimulated units after the combined blockade of D1 and D2 receptors. Responses to cocaine were mimicked by iontophoretic procaine (0-40 nA), a short-acting local anaesthetic with minimal effect on DA uptake. Procaine-induced inhibitions occurred at the same low currents, had a similar time-course, and were also strongly dependent on basal discharge rate. Our data support the existence of a DA-independent mechanism for the action of cocaine involving a direct interaction with Na+ channels. Although further studies are required to clarify this mechanism and its interaction with other pharmacological and behavioural variables, a direct interaction with Na+ channels may contribute to changes in neuronal activity induced by self-injected cocaine, thereby playing a role in mediating the psychoemotional and behavioural effects of this drug.     

利用胶原构建皮肤组织工程支架的研究

目的 利用胶原海绵膜构建皮肤组织工程支架。方法 从大鼠鼠尾肌腱中提取胶原。胶原和6-硫酸软骨素冷冻干燥，重度脱水及戊二醛交联后制得胶原海绵膜。将其行扫描电镜观察；并埋藏在Balb／c小鼠皮下，术后定期取出观察组织学变化。将成纤维细胞种植于该支架上。结果 构建的支架孔径均匀，孔径在50～100μm；具有良好的组织相容性，并逐渐降解。成纤维细胞于其上生长良好。结论 该支架适合成纤维细胞生长，可作为组织工程皮肤的支架。