灯盏生脉胶囊抗大鼠局灶性脑缺血-再灌注损伤作用研究

目的 研究灯盏生脉胶囊抗大鼠局灶性脑缺血 再灌注损伤的作用。方法将58只雄性SD大鼠随机分为4组，即假手术组10只，0.9%氯化钠溶液组和灯盏生脉胶囊高、低剂量组各16只。假手术组行假手术，其他3组制作大脑中动脉缺血 再灌注模型。假手术组和0.9%氯化钠溶液组于术前72，48，24，0.5 h及术后22 h灌胃给予0.9%氯化钠溶液，灯盏生脉胶囊低、高剂量组在相同时点分别按0.36和0.72 g·kg 1剂量灌胃给予灯盏生脉胶囊内容物悬浊液4 mL；各组大鼠均在最后一次给药后2 h处死，于脑缺血2 h和再灌注2和22 h分别进行神经行为学评分，再灌注22 h后测定大鼠血清乳酸脱氢酶活性及脑梗死体积百分比，观察脑组织病理学改变。结果再灌注2 h后，灯盏生脉胶囊高剂量组神经行为学评分［(1.75±0.68)分］较0.9%氯化钠溶液组［(2.31±0.79)分］明显降低（P＜0.05）；再灌注22 h后，灯盏生脉胶囊高剂量组神经行为学评分［(1.63±0.89) 分］明显低于0.9%氯化钠溶液组［(2.38±1.09)分］（P＜0.05）。再灌注22 h后，灯盏生脉胶囊高剂量组血清乳酸脱氢酶活性［(1 331.89±366.85) U·L 1］明显低于0.9%氯化钠溶液组［(1 799.56±325.92) U·L-1］（P＜0.05），灯盏生脉胶囊高剂量组脑梗死体积百分比［(9.73±5.66)%］亦明显低于0.9%氯化钠溶液组［(18.31±6.96)%］（P＜0.05）。灯盏生脉胶囊低、高剂量组鼠脑组织缺血周围区细胞周围水肿、神经元变性和间质破坏较其他各组轻。结论灯盏生脉胶囊有一定的抗大鼠局灶性脑缺血 再灌注损伤作用，作用机制可能在于抑制缺血半暗带恶化。高剂量作用更明显。     

褐藻多糖硫酸酯对大鼠脑缺血再灌注损伤后的神经保护作用

目的探讨褐藻多糖硫酸酯（FPS）对脑缺血再灌注损伤的神经保护作用及可能机制。方法将48只大鼠按照随机数字表法分为假手术组、大脑中动脉栓塞（MCAO）组、FPS低剂量组（MCAO＋FPS 50 mg/kg）、FPS高剂量组（MCAO＋FPS 100 mg/kg）,每组12只。假手术组大鼠只分离右侧颈总动脉及颈内外动脉,MCAO组、FPS低剂量组、FPS高剂量组采用线栓法制作大脑中动脉栓塞模型,缺血2 h后再灌注后FPS低、高剂量组予鼠尾静脉注射FPS50、100 mg/kg。对所有大鼠按照Longa标准进行神经功能评分;采用TTC法测量脑梗死体积;蛋白印迹法测定Nrf2蛋白的表达。结果与MCAO组比较,FPS低、高剂量组大鼠神经功能评分明显降低、转录因子NF-E2相关因子2（NF-E2-related factor 2,Nrf2）蛋白表达明显增高,FPS高剂量组神经功能评分、Nrf2蛋白表达改变得更为显著（P〈0.05,P〈0.01）;FPS高剂量组大鼠脑梗死体积明显减小（P〈0.01）。结论 FPS通过提高Nrf2的表达缩小脑梗死体积,对脑缺血再灌注损伤大鼠起到神经保护作用。     

依达拉奉对脑缺血再灌注损伤

［摘要］目的研究自由基清除剂依达拉奉对脑缺血 再灌注损伤大鼠的抗细胞凋亡和神经保护作用。方法采用线栓法制备大鼠大脑中动脉缺血 再灌注模型。将SD大鼠分为假手术组、再灌注组和依达拉奉组。再灌注组和依达拉奉组按再灌注2，6，12，24，48 h分为5个亚组。依达拉奉组在缺血2 h后解除栓塞，给予依达拉奉3 mg·kg-1静脉注射，首次给药24 h后相同剂量再次给药。再灌注组给予0.9%氯化钠溶液，给药剂量、时间和方法同依达拉奉组。应用硫代巴比妥酸(TBA)比色法检测各组血清丙二醛(MDA)浓度，免疫组化染色及原位细胞凋亡检测法(TUNEL法)测定脑组织bcl 2蛋白表达和凋亡细胞数，并测量各组脑梗死体积。结果依达拉奉组再灌注后6，12，24，48 h的梗死体积、血清MDA浓度、TUNEL阳性细胞数均明显小于再灌注组（均P＜0.05），各时间点bcl 2蛋白表达均高于再灌注组（P＜0.01）。结论依达拉奉可以降低羟自由基水平，对抗细胞凋亡，对脑缺血 再灌注损伤大鼠有明显保护作用。     

脑心通对大鼠神经功能评分及梗死体积的影响

目的：观察脑缺血再灌注损伤后SD大鼠不同时间点神经功能评分及梗死体积的变化,探讨脑心通对它们的影响。方法线栓法制作大鼠大脑中动脉缺血（Middle cerebral artery occlusion,MCAO）模型,选用健康雄性SD大鼠68只,随机分成四组：假手术组;MCAO模型组;MCAO＋脑心通胶囊低剂量组;MCAO＋脑心通胶囊高剂量组。每组再按照脑缺血再灌注后第3d、第7d、第14d和第21d 4个不同时间点进行神经功能评分及2,3,5-氯化三苯基四氮唑（2,3,5-Triphenyltetrazolium chloride,TTC）染色测定大鼠脑缺血再灌注后梗死体积。结果除假手术组外,其余三组实验鼠在缺血再灌注后4个不同时间点之间比较神经功能评分具有差异（P〈0.01）;不同剂量脑心通组与MCAO模型组比较差异有统计学意义（P〈0.05）;但脑心通高、低剂量组之间比较无明显差异（P〉0.05）。 TTC染色发现缺血再灌注后第7d各组大鼠脑梗死体积最大;不同时间点之间比较差异有统计学意义（P〈0.05）;不同剂量的脑心通实验组与模型组之间比较,差异具有统计学意义（P〈0.01）。结论脑心通胶囊对改善缺血再灌注损伤后实验鼠神经功能评分和减轻梗死体积有促进作用,但对神经功能评分的影响与药物剂量不相关。     

口服PTD-SOD对大鼠局灶性脑缺血再灌注损伤的保护作用

目的探讨口服灌胃PTD．SOD对大鼠局灶性脑缺血再灌注（cerebral ischemia-reperfusion,CIR）引起的脑损伤的保护作用及机制。方法采用线栓法制作大鼠大脑中动脉闭塞再灌注模型（middl ecerebral artery occlusion,MCAO）,缺血2h,再灌注24h。于缺血前一周开始灌胃给予PTD-SOD10,20,40mg·kg-1,2次d-1。对术后MCAO大鼠进行神经行为学特性观察并评分;TTC染色法测量大鼠脑梗死面积;生化法检测大鼠脑组织匀浆中s0D、GSH-Px、CAT的活力和MDA、NO的含量。HE染色观察大鼠脑组织病理改变。结果PTD．SOD可明显改善大鼠神经行为,减少脑梗死面积,提高脑内SOD、GSH-Px、CAT的活性,降低脑组织中MDA及N0含量,减轻脑组织病理改变（P〈0．05或P〈O．01）。结论PTD．SOD可减轻CIR神经细胞损伤,其机制可能与其能抗自由基损伤,减轻NO神经毒性有关。     

表儿茶素对大鼠脑缺血再灌注损伤的保护作用及抗氧化能力的影响#br#

摘要：目的　研究表儿茶素（EC）对大鼠脑缺血再灌注后的保护作用及抗氧化能力的影响。方法　取90只雄性清洁级SD大鼠,按随机数字表法分为6组,每组15只,分别为假手术组（Sham组）、CIRI模型组（I/R组）、EC 5 mg/kg组、EC 10 mg/kg组、EC 20 mg/kg组、依达拉奉（ED）3 mg/kg组。除Sham组外,其余组采用线栓法构建大脑中动脉栓塞（MCAO）模型,缺血2 h,于再灌注后0、12、24 h给药,12、24 h进行Longa法神经功能损伤评分,36 h取脑行氯化三苯基四氮唑（TTC）染色检测缺血侧脑组织梗死面积,HE染色观察缺血侧脑组织形态学变化,比色法检测血清和缺血侧脑组织中的丙二醛（MDA）含量、超氧化物歧化酶（SOD）活力以及脑组织中谷胱甘肽过氧化物酶（GPx）的活力。结果　与Sham组比较,I/R组神经功能损伤评分升高,梗死面积百分比增大（P＜0.05）,组织形态明显改变,血清和脑组织中MDA含量增多、SOD活力降低（P＜0.05）,脑组织中GPx活力降低（P＜0.05）。与I/R组比较,EC 10 mg/kg、EC 20 mg/kg、ED 3 mg/kg组神经功能损伤评分降低,梗死面积百分比减少（P＜0.05）,组织损伤有所改善,血清和缺血侧脑组织中MDA含量减少、SOD活力升高,EC 10 mg/kg、EC 20 mg/kg、ED 3 mg/kg组脑组织中GPx活力升高（P＜0.05）。结论　EC能提高脑抗氧化能力,对抗大鼠脑缺血再灌注损伤,且在一定范围内呈剂量依赖性。     

钙蛋白酶抑制剂calpeptin对大鼠局灶性脑缺血再灌注损伤的影响

目的 应用大脑中动脉闭塞(MCAO)模型,研究钙蛋白酶抑制剂calpeptin对大鼠局灶性脑缺血再灌注损伤的影响.方法 32只SD大鼠随机分为4组,每组8只.单纯缺血组(I组):大脑中动脉闭塞2h;calpeptin组(C组)大鼠在2h的大脑中动脉闭塞前30min经侧脑室注射calpeptin50μg;二甲基亚砜(DMSO)组(D组)大鼠在2h的大脑中动脉闭塞前30min经侧脑室注射DMSO 5μL;对照组(S组)不闭塞大脑中动脉,其余手术操作同其他组.各组大鼠在2h的缺血后均再灌注48h.分别在再灌注12、24、48h行神经功能学评分,在再灌注48h后应用TTC染色法测定脑梗死体积.结果 S组大鼠在再灌注后各时间点的神经功能学评分与术前相比无明显变化,手术后48h未发现梗死灶.C组大鼠各时间点的神经功能学评分及再灌注48h脑梗死体积均低于I组和D组(P＜0.05),而I组和D组之间无显著性差异(P＞0.05).结论 缺血前应用钙蛋白酶抑制剂可以显著减轻大鼠局灶性脑缺血再灌注损伤.     

依那普利对肾陛高血压大鼠脑缺血再灌注后脑组织基质金属蛋白酶-2、-9表达的影响

目的 通过观察血管紧张素转化酶抑制剂（ACEI）依那普利对肾性高血压大鼠脑缺血再灌注后脑组织基质金属蛋白酶（MMP）-2和MMP-9表达的影响,探讨依那普利的脑保护机制.方法 Wistar雄性大鼠28只,随机分为高血压组和正常血压组.前组采用狭窄肾动脉方法建立肾性高血压大鼠模型,再随机分为依那普利组（Y）和缺血再灌注组（HIR）,分别采用依那普利2 mg/kg和0.9%氯化钠注射液进行灌胃治疗;正常血压组再分为假手术组（N）和缺血再灌注组（IR）,分别行假手术和缺血再灌注处理.采用大脑中动脉线栓法造成大脑缺血再灌注模型,缺血时间为2 h,再灌注22 h.采用免疫组织化学技术检测脑组织MMP-2和MMP-9的表达,用图象分析仪测定灰度值.结果 与N组比较,IR组MMP-2、MMP-9灰度值明显增高（P＜0.01）;与IR组比较,HIR组MMP-2、MMP-9灰度值增高（P＜0.05）;与HIR组比较,Y组MMP-2、MMP-9灰度值减少（均P＜0.01）.结论 高血压加重缺血再灌注后脑组织MMP-2和MMP-9的表达;依那普利能明显抑制脑组织MMP-2和MMP-9的表达.     

二巯基丙磺酸钠对大鼠脑缺血再灌注损伤的影响

目的探讨二巯基丙磺酸钠对大鼠短暂性局灶性脑缺血再灌注损伤的影响。方法雄性SD大鼠随机分为假手术组(Sham组)、缺血再灌注组(即生理盐水组,简NS组)、二巯基丙磺酸钠组(DMPS组)。参照ZeaLonga等方法栓塞右侧大脑中动脉制作短暂性局灶性脑缺血再灌注模型(tMCAO),除Sham组仅施行手术而不进行大脑中动脉栓塞术外,其余大鼠均接受大脑中动脉栓塞术,缺血2h时退出栓线实现再灌注,以出现提尾悬空时左前肢屈曲内收,视为模型成功标准。在再灌注24h后取全脑制成10%匀浆,光谱比色法测定MAD含量、SOD和GSH-Px活力。结果①tMCAO大鼠神经行为缺陷评分:各组大鼠分别于再灌注24h按照Julio神经行为缺陷评分法进行神经行为缺陷评分,各缺血组均有不同程度的神经行为缺陷,药物治疗后,能不同程度的改善缺血再灌注大鼠神经行为缺陷(与NS组比较,P<0.05);②tMCAO大鼠梗死面积百分比测定:缺血再灌注脑组织均有不同程度的梗死现象,经二巯基丙磺酸钠治疗后,能不同程度的的减少缺血再灌注大鼠脑组织的梗死面积百分比(与NS组比较,为P<0.05);③HE染色光镜检查:tMCAO大鼠缺血再灌注后脑组织坏死范围增大,存活细胞减少,坏死严重,DMPS组能改善梗塞区神经细胞的变性程度;④与缺血再灌注组(NS)比较,DMPS组不完全性全脑缺血再灌注损伤后,大鼠MAD含量显著下降(P<0.01),SOD活性和GSH-Px活性均显著提高(均为P<0.01)。结论二巯基丙磺酸钠能显著减少脑缺血再灌注后MAD含量,提高SOD和GSH-Px活性;改善神经行为缺陷和减少梗死面积百分比,减轻神经细胞坏死。     

后适应对大鼠脑缺血神经细胞凋亡和内皮型一氧化氮合酶与诱导型一氧化氮合酶的影响

目的：研究缺血耐受（后适应和预适应）对脑缺血再灌注神经细胞凋亡及内皮型一氧化氮合酶（eNOS）和诱导型一氧化氮合酶（iNOS）蛋白表达的影响。方法：50只雄性SD大鼠随机分为假手术组（sham）、脑缺血再灌注模型组（MCAO）、预适应组（MCAO＋preconditioning）、后适应组（MCAO＋postconditioning）和尼莫地平组（MCAO＋nimodipine），每组10只大鼠。预适应组大鼠在MCAO前24h，双侧颈总动脉夹闭2min，再灌注20min，循环两次。后适应组大鼠在脑缺血再灌注开始时，再灌注20s，栓塞20s，循环3次。大鼠大脑中动脉阻断1．5h，再灌注24h。用TUNEL法和免疫组化染色法分别检测缺血半暗带凋亡细胞和iNOS、eNOS蛋白表达。结果：与MCAO组比较，后适应组大鼠脑缺血半暗带的凋亡细胞显著减少，eNOS蛋白表达显著增加，iNOS蛋白表达显著减少，差异有统计学意义（P〈0．05）。结论：缺血后适应能诱导脑缺血耐受，对脑缺血／再灌注损伤产生保护作用，其保护作用与促进eNOS蛋白的表达，抑制iNOS蛋白的表达有关。     

白藜芦醇预处理对大鼠局灶性脑缺血/再灌注损伤的神经保护作用

目的探讨白藜芦醇(resveratrol,Res)预处理对大鼠局灶性脑缺血/再灌注损伤的神经保护作用。方法♂SD大鼠45只,随机分为假手术组(Sham)、缺血/再灌注组(I/R)和白藜芦醇预处理缺血/再灌注组(Res+I/R),每组15只。采用线栓法行大脑中动脉阻断前脑血90min,拔出栓线实现再灌注。Res+I/R组缺血前30min腹腔注射白藜芦醇(30mg·kg-1)。再灌注后24h,用2,3,5-氯化三苯基四氮唑(TTC)染色显示梗死范围;再灌注72h后利用Morris水迷宫检测脑缺血后大鼠空间学习记忆能力,用电生理学方法检测白藜芦醇对脑缺血大鼠突触可塑性的影响。结果白藜芦醇预处理可以明显缩小脑梗死体积并改善大鼠的行为学障碍(P<0.05),在Morris水迷宫实验中Res+I/R组大鼠逃避潜伏期明显短于I/R灌注组(p<0.05),高频刺激后,I/R组海马CA1区NMDA受体依赖性的长时程增强(LTP)诱导障碍(n=5),Res+I/R组海马CA1区可诱导稳定的LTP(n=5)。结论缺血前30min白藜芦醇预处理对局灶性脑缺血/再灌注损伤具有神经保护作用,可以缩小脑梗死体积,并对大鼠的学习与记忆能力具有改善作用。     

Therapeutic time window for treatment of focal cerebral ischemia reperfusion injury with XQ-1h in rats

Pervious experimental studies have shown that XQ-1h has beneficial neuroprotective effect in the cerebral ischemia reperfusion injury. However, the therapeutic time window for treatment of focal cerebral ischemia reperfusion injury with XQ-1h is not clear. Under chloral hydrate anesthesia, transient focal cerebral ischemia was induced in rats by 2h of middle cerebral artery occlusion (MCAO), followed by 24h of reperfusion. Saline as vehicle or XQ-1h at the doses of 31.2, 15.6 and 7.8 mg/kg i.v. was administered at 0.5, 1, 2, 3h after induction of ischemia. Subsequently, 24h after MCAO brain edema, infarct volume, neurological deficits and cerebral blood flow were evaluated. Administrations of XQ-1h at the doses of 31.2mg/kg at 0.5, 1, and 2h after reperfusion of MCAO significantly reduced infarct rate (%) by 75.6% (5.2 ± 1.7), 66.2% (7.2 ± 1.9), and 47.9% (11.1 ± 1.2), respectively. XQ-1h (31.2mg/kg) treatment, 0.5, 1, and 2h after reperfusion produced significant improvement in neurological score compared to vehicle-treated group (P<0.01). Administrations of XQ-1h at the doses of 31.2mg/kg and 15.6 mg/kg at 0.5, 1, and 2h after reperfusion of MCAO significantly increased cerebral blood flow (mv) by 16.9 ± 1.9, 11.7 ± 1.3, 9.5 ± 1.0, respectively (P<0.01). In conclusion the therapeutic time window of XQ-1h for cerebral ischemia reperfusion injury is within 2h. Interestingly, we also discovered that the therapeutic time window of XQ-1h is deeply related with the activity of scavenging oxidative stress products. Further studies need to be conducted more drug combination therapy programs in order to assess the potential clinical application of XQ-1h.     

Effects of sodium beta-aescin on expression of adhesion molecules and migration of neutrophils after middle cerebral artery occlusion in rats.

AIM: To investigate the effects of sodium beta-aescin on neutrophil migration and expression of adhesion molecules (ICAM-1 and E-selectin) after middle cerebral artery occlusion (MCAO) in rats. METHODS: Rats were pretreated with sodium beta-aescin for 7 d and then subjected to cerebral ischemia/reperfusion (I/R) injury induced by an MCAO. After a 2-h ischemia and a 24-h reperfusion, the infarct volume and neurological deficit were determined by the method of TTC staining and the Longa's score. The effect of sodium beta-aescin on the migration of neutrophils was evaluated by measuring the activity of myeloperoxidase (MPO) enzyme. The expressions of adhesion molecules were determined by immunohistochemistry and Western blot. RESULTS: Sodium beta-aescin significantly reduced the cerebral infarct volume and ameliorated the neurological deficit (P<0.05 or P<0.01). The MPO activity and the expressions of ICAM-1 and E-selectin in the vehicle-treated rats were increased significantly (P<0.01) after cerebral I/R. After treatment with sodium beta-aescin, the enzymatic activity of MPO and the expressions of these adhesion molecules were significantly reduced compared with the vehicle-treated group (P<0.05 or P<0.01). CONCLUSION: Sodium beta-aescin can attenuate brain injury, down-regulate the protein expressions of ICAM-1 and E-selectin, and reduce the migration of neutrophils after cerebral I/R.     

阿司匹林预处理对大鼠脑缺血再灌注损伤的神经保护作用

目的:观察阿司匹林(ASA)预处理对大鼠局灶性脑缺血再灌注损伤(I/R)的神经保护作用,并对其作用机制进行探讨。方法:复制大鼠大脑中动脉缺血再灌注模型,分别采用TTC染色法、神经功能缺损评分法观察ASA预处理对大鼠脑梗死体积和神经功能评分的影响,以及对脑组织中超氧化物歧化酶(SOD)、丙二醛(MDA)和血前列环素I2(PGI2)/血栓素A2(TXA2)的影响。结果:ASA能够降低I/R大鼠脑梗死体积和神经功能评分,增加脑组织中SOD的活性,降低MDA的含量,升高血PGI/TXA2的比值。结论:ASA对大鼠脑缺血再灌注损伤有一定保护作用,其作用机制与增加脑组织中SOD的活性、降低MDA的含量、提高血PGI2/TXA2的比值有关。     

香芹酚通过降低一氧化氮水平防治大鼠脑缺血/再灌注损伤的研究

目的研究香芹酚通过降低一氧化氮水平对大鼠脑缺血/再灌注(CI/R)损伤的保护作用。方法制做大鼠脑中动脉闭塞(MCAO)2 h后再灌注24 h模型,将大鼠随机分为5组:假手术组,模型组,香芹酚低、中、高剂量组。实验终末,评价神经功能及脑梗死面积,并检测脑组织中NO的含量。结果与模型组相比,香芹酚组神经功能学评分显著下降(P<0.05),大鼠脑梗死面积亦显著减少(P<0.05)。模型组中再灌注24 h后,大鼠缺血侧和自身对照侧皮质及海马NO的含量异常升高(P<0.05);香芹酚组缺血侧和自身对照侧皮质及海马NO的含量明显降低(P<0.05)。结论香芹酚对大鼠CI/R损伤有保护作用,其机制与降低CI/R损伤时的脑组织中的NO水平可能相关。     

Neuroprotective effects of scutellarin on rat neuronal damage induced by cerebral ischemia/reperfusion

AIM: To investigate the neuroprotective effect and mechanisms of scutellarin, a flavonoid extracted from Erigeron breviscapus Hand Mazz, against neuronal damage following cerebral ischemia/reperfusion. METHODS: Rats were pretreated ig with scutellarin for 7 d and then subjected to cerebral ischemia/reperfusion (I/R) injury induced by a middle cerebral artery occlusion (MCAO). The infarct volume and neurological deficit were determined by TTC staining and Longa's score. The permeability of the blood-brain barrier was evaluated by measurement of the Evans blue (EB) content in the brain with a spectrophotometer. The total NOx content was determined. Nitric oxide synthase (NOS) isoforms (iNOS, eNOS, nNOS) and the key angiogenic molecules, vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF), were detected by Western blotting. RESULTS: Scutellarin significantly reduced infarct volume (P<0.05 or P<0.01), ameliorated the neurological deficit and reduced the permeability of the blood-brain barrier (BBB) (P<0.05). When rats were pretreated with scutellarin (50 or 75 mg/kg), upregulation of eNOS expression and downregulation of VEGF, bFGF, and iNOS expression was observed, whereas scutellarin had no effect on nNOS expression. CONCLUSION: Scutellarin has protective effects for cerebral injury through regulating the expression of NOS isoforms and angiogenic molecules.     

Neuroprotection of total steroid saponins from Dioscorea zingiberensis C.H.Wright against transient focal cerebral ischemia-reperfusion injury in rats via its anti-inflammatory and anti-apoptotic effects

OBJECTIVE The total steroid saponins(TSSN) isolated from Dioscorea zingiberensis C. H. Wright(D.zingiberensis) has shown a variety of beneficial bioactivities. However, there are no reports about the neuroprotective effects of the TSSN until now. Therefore, we explored the neuroprotective effects of TSSN on rats against transient focal cerebral ischemia-reperfusion(I/R) and the underlying mechanisms. METHODS The healthy adult Sprague-Dawley rats were randomly assigned into six groups. After pre-treatment with the TSSN intragastrically for six days, the rats were subjected to the ischemia injury by the surgery of middle cerebral artery occlusion(MCAO) for 90 min. Some indexes were evaluated and detected. RESULTS As compared to the I/R group, TSSN group of rats, especial y given the 30 mg·kg~(-1) of TSSN, not only marked reduction in the neurological deficit scores, cerebral infarct volume, and brain edema, but also an increase in neuron survival(Nissl bodies) in the hippocampal cornuammons 1(CA1) and cortex hemisphere of the ipsilateral ischemia. At the same time, the inflammatory cytokines in serum induced by MCAO were significantly alleviated by the TSSN pre-administration. What′s more, the increase of caspase-3 was evidently reduced in the CA1 and cortex of the hemisphere injured brain. Final y, the down-regulating anti-apoptotic Bcl-2 and up-regulating pro-apoptotic Bax proteins were obviously suppressed. CONCLUSION TSSN plays a potential neuroprotective role against a severe injury induced by transient focal cerebral ischemic reperfusion in a rat experimental model, and this role may be mediated by its antiinflammatory and anti-apoptotic actions.     

蕲蛇酶对大鼠脑缺血再灌注损伤的保护作用

目的进一步探讨蕲蛇酶对大鼠脑缺血再灌注损伤的保护作用及其机制。方法采用线栓法制备大鼠大脑中动脉闭塞后再灌注(MCAOR)模型,观察大鼠MCAOR时神经功能状态,同时,TTC染色测脑梗死面积,电镜观察脑血管超微结构,放免法测定血浆血栓素B2及6-酮前列腺素F1α含量。用药组分别在缺血即刻或再灌注即刻静脉给药,观察比较模型组和蕲蛇酶所有给药组之间上述指标的变化。结果蕲蛇酶所有给药组都能有效改善MCAOR大鼠神经功能缺失症状,明显减小梗死灶,调节血栓素B2/6-酮前列腺素F1α,保护血管内皮细胞,维持微血管正常结构。结论蕲蛇酶对脑缺血再灌注损伤保护作用可能与维持血栓素B2/6-酮前列腺素F1α平衡、改善脑微循环有关。     

Effect of active components group of Xiaoxuming decoction on cerebral mitophagy after focal cerebral ischemia/reperfusion injury

OBJECTIVE To explore the effect of the active components group of Xiaoxuming decoction(XXMD)on mitophagy after cerebral ischemia/reperfusion(I/R) in MCAO rats. METHODS Male Sprague-Dawley rats were randomly divided into 4 groups: sham group, XXMD and rapamycin group. The I/R models were induced by middle cerebral artery occlusion. The effects of XXMD and rapamycin on behavioral study of focal cerebral I/R in rats were measured by Zea-Longa′ s level standard scoring method and neurological defect score(m NSS). The volume percentage and hemispheric swelling of rat cerebral infarction were detected by TTC staining. Proteins in the mitophagy were detected by Western blotting. RESULTS The behavioral results showed that compared with the model group, XXMD decoction and rapamycin could significantly alleviate the neurological deficit scores after I/R, decreased the m NSS score(P<0.05) of the MCAO rats. TTC results showed that XXMD and rapamycin could significantly reduce the percentage of infarct volume and hemispheric swelling(P<0.05) of MCAO rats. Compared with the model group, the XXMD could significantly inhibit the expression of P62, Beclin-1 and BNIP3 and decrease LC3-Ⅱ/LC3-Ⅰ ratio(P<0.05, P<0.01) in isolated cerebral mitochondrial proteins at 24 h after cerebral I/R. However, the expression of P62,Beclin-1, BNIP3(P<0.05, P<0.01) and the ratio of LC3-Ⅱ/LC3-Ⅰin purified mitochondrial proteins can be significantly elevated by XXMD at 96 h after cerebral I/R. CONCLUSION Autophagy and mitophagy were activated at 24 h after cerebral I/R, and then it was inhibited. XXMD can protect mitochondrial structures, regulate the level of mitophagy at different points after cerebral I/R, inhibit the excessive activation of mitophagy in 24 h after I/R, and improve the level of mitophagy in the later stage of I/R.