柔肝通络汤联合依达拉奉对脑梗死模型大鼠脑可塑性物质及结构影响作用

目的:分析柔肝通络汤联合依达拉奉对脑梗死模型大鼠脑可塑性物质及结构影响。方法:选取SPF级Wistar雄性大鼠60只,随机数字表法将大鼠分成三组,正常组、模型组与观察组,每组各20只。除正常组其他各组大鼠制备脑梗死模型,给药后7天随机选取8只大鼠,给药后21天选取剩余大鼠,断颈处死后选取脑组织内缺血中心位置冠状脑片,行HE染色,免疫检测大鼠冠状脑片内突触素(SY38)及神经微丝蛋白(NFP-200)表达状况。结果:给药7天、21天时模型组大鼠SY38、NFP-200表达量比正常组下降,观察组比模型组上升,差异均有统计学意义(P<0.05)。结论:依达拉奉结合柔肝通络汤可使脑梗死大鼠脑组织内NFP-200、SY38表达量升高,降低神经细胞和毛细血管缺血性受损,加速突出超微结构功能变强。

Correlation of free radical level and apoptosis after intracerebral hemorrhage in rats

Objective To investigate the correlation of perihematomal free radical level and neuronal apoptosis following the intracerebral hemorrhage （ICH）. Methods Animals were randomly divided into 4 groups： sham operation group, model group, 1 mg/kg edaravone group, and 3 mg/kg edaravone group. Each group was then divided into seven subgroups, in which the rats were correspondingly killed at 6 h, 12 h, 24 h, 48 h, 72 h, 7 d or 14 d （n = 1 in each subgroup of the sham group, and n = 6 in each subgroup of the other 3 groups）. By Horseley-Clarke technique, autoblood （80 μL） were administered into the left caudate putamen of SD rats in a double administration-withdrawal way. Rats in the sham group were needled in but not administered with autoblood. The ICH model was then evaluated by Bederson＇s scale. Around the hematoma, the levels of malonaldehyde （MDA） and hydroxyl radical were tested by spectrophotometer, and the process of apoptosis was tested by terminal deoxynucleofidyltransferase-mediated dUTP-biotin nick end labeling （TUNEL） method. Results （1） ICH significantly increased the levels of MDA and hydroxyl radicals. Significant differences in MDA and hydroxyl radical contents were observed among the four groups. （2） In the sham group, a small number of TUNEL-positive cells were found. In the other three groups, the TUNEL-positive cells were observed at 6 h, increased significantly at 24 h, and reached peak level at 3 d, then fell profoundly at 7 d, but remained detectable at 14 d. （3） The positive correlation existed between apoptosis and free radical level （r = 0.2003）, and existed between apoptosis and MDA content （r = 0.6563） in the brain. Conclusion Post-hemorrhagic apoptosis was related to the production of free radicals, indicating that the elevated free radicals following the ICH could induce neuron and glial cell apoptosis.

Edaravone inhibits apoptosis caused by ischemia/reperfusion injury in a porcine hepatectomy model

AIM： TO investigate the effect of E3-methyl-l-phe- nyl-2-pyrazolin-5-one （Edr） on hepatic ischemia-reper- fusion （I/R） injury and liver regeneration in a porcine hepatectomy model. METHODS： One hour ischemia was induced by occlud- ing the vessels and the bile duct of the right and median lobes. A 40% left hepatectomy was performed after re- perfusion. Six animals received Edr （3 mg/kg per hour） intravenously and six control animals received saline just before reperfusion. Remnant liver volume, hemody- namics, aspartate aminotransferase （AST）, alanine ami- notransferase, lactate dehydrogenase and lactic acid, were compared between the groups. The expression of transforming growth factor-β （TGF-β1） and toll-like receptor （TRL） mRNA in hepatic tissues was examined using reverse transcription polymerase chain reaction. Apoptosis was demonstrated by terminal deoxynucleo- tidyl transferase dUTP nick end labeling （TUNEL） stain- ing, respectively. RESULTS： Serum AS-I- （P = 0.029）, and toll like recep- tor 4 level （P = 0.043） were significantly lower after 3 hin animals receiving Edr. In addition, TUNEL staining in Edr-treated pigs showed significantly fewer hepatocytes undergoing apoptosis compared with control pigs. After 1 mo, all factors were non-significantly different between the two groups. CONCLUSION： Edr is considered to reduce hepatic injury in the early stage of I/R injury in a porcine model.

丹红注射液联合依达拉奉在脑梗死患者神经功能恢复中的临床研究

目的:探讨丹红注射液联合依达拉奉在脑梗死患者神经功能恢复中的临床疗效。方法:随机抽取2012年3月至2015年1月我院收治的128例脑梗死患者的临床资料,按照随机数字表法将患者均分为实验组和对照组,实验组和对照组患者例数均为64例。患者均进行常规营养支持治疗的同时,加用抗血小板聚集药、脑细胞活化药等,对照组患者采取依达拉奉联合灯盏花素治疗;实验组患者采取丹红注射液、灯盏花素联合依达拉奉治疗,观察两组患者神经功能缺损评分(NIHSS)、神经功能缺损程度评分量表(NDS评分)、临床疗效、不良反应发生情况。结果:实验组患者神经功能缺损评分、NDS评分在治疗后均明显低于对照组和治疗前。对照组患者神经功能缺损评分、NDS评分在治疗后均明显低于治疗前。实验组患者治疗总有效率(90.63%)高于对照组(73.44%),两组患者均未发生严重不良反应。两组患者治疗后IL-6、IL-8、TNF-α、hs CRP、MMP-9水平均明显低于治疗前;实验组患者治疗后IL-6、IL-8、TNF-α、hs CRP、MMP-9水平明显低于对照组。两组患者治疗后TG、TC、LDL-C水平均低于治疗前;实验组患者治疗后TG、TC、LDL-C水平低于对照组,两组患者治疗前后HDL-C水平变化差异无统计学意义。两组患者不良反应发生率比较,差异无统计学意义。结论:丹红注射液、灯盏花素联合依达拉奉在脑梗死患者神经功能恢复中的应用价值较高,可改善患者NIHSS、NDS评分,提高临床疗效,无明显不良反应。

Protective effects of edaravone against cobalt chloride-induced apoptosis in PC12 cells

Objective To investigate the neuroprotective effects of edaravone （Eda） on cobalt chloride （CoCl2）-induced oxidative stress and apoptosis in cultured PC12 cells as well as the underlying mechanisms. Methods PC12 ceils impaired by COCl2 were used as the cell model of hypoxia. MTT （methyl thiazolyl tetrazolium） was used to assay the viability of the PC12 ceils exposed to Eda with gradient concentrations; Hochest 33258 stain assay was used to analyze the apoptosis ratio of the PC12 cells; Bcl-2 and Bax protein levels in PC12 cells were examined by western blotting. ROS level, the mitochondrial transmembrane potential and caspase-3 activity in each group were detected by spectrofluorometer. Results COCl2 treatment caused the loss of cell viability in PC12 cells, which was associated with the elevation of apoptotic rate, the formation of ROS and the disruption of mitochondrial transmembrane potential. COCl2 also significantly induced the upregulation of Bax/Bcl-2 ratio and the activation of caspase-3. In contrast, Eda significantly reversed these phenotypes, with its maximum protective effect at 0.1 μmol/L. Conclusion These results indicated that Eda could protect PC12 cells from CoCl2-induced cytotoxicity, and this protection might be ascribed to its anti-oxidative and anti-apoptotic activities.

Electrophysiological study on differentiation of rat bone marrow stromal stem cells into neuron-like cells in vitro by edaravone

Objective： To explore the electrophysiological proper- ties of differentiation of rat bone marrow-derived stromal stem cells （rBMSCs） to neuron-like cells in vitro by edaravone, a new type of free radical scavenger. Methods： Stromal stem cells were separated from rat bone marrow with Ficoll-Paque reagent and expanded in different culture medium in vitro, rBMSCs were induced by edaravone containing serum-free L-DMEM. Morphologic observation and Western blot analysis including the expression of Nav1.6, Kvl.2, Kv1.3, Cav1.2 were performed, and whole patch-clamp technique was used. Results： Cyton contraction and long processes were shown in differentiated stromal stem cells. Navl.6, Kvl.2, Kv 1.3 and Cav 1.2 were expressed in both differentiated and undifferentiated ceils. However, the expression of channel proteins in differentiated cells was up-regulated. Consistently, their resting potential and outward currents were also enhanced in the differentiated cells, which was especially significant in the outward rectifier potassium current. Conclusion： In vitro, neuron-like cells derived from rBMSCs, induced by edaravone, possess electrophysiological properties of neurons.

Synthesis of two metabolites of edaravone

Edaravone glucuronate and edaravone sulfate,two metabolites of edaravone,were synthesized in improved yields. Edaravone glucuronate was synthesized by conjugation of edaravone with glucuronolactone through a series of reactions.Edaravone sulfate was synthesized through sulfonation of edaravone.Compared with the reported methods in the literature,the yields of both metabolites with the improved methods were much higher.