摘要
目的:观察黄芪多糖对大鼠脑出血后神经细胞凋亡及核因子κB表达的影响。方法:实验于2006-01/06在南京医科大学中心实验室完成。①选用健康雄性SD大鼠69只,按随机数字表法分为3组:假手术组、模型组与治疗组,每组23只,每组分别于术后6h,1,3,5d取5只用于病理切片制作,于术后3d每组各取3只用于电镜检查。②模型组与治疗组均采用Ⅶ型胶原酶诱导脑出血模型,假手术组以等量生理盐水代替Ⅶ型胶原酶;治疗组于造模术后2h给予黄芪多糖(美国泛华医药公司Pharmagenesis提供,批号8k01101)25mg/kg腹腔注射,1次/d;其余两组在同样时间点给予2mL生理盐水腹腔注射。③用原位末端标记技术和免疫组化方法检测血肿周围神经凋亡和核因子κB表达的动态变化。④透射电镜观察术后3d血肿周围神经元超微结构的变化。结果:大鼠69只均进入结果分析。①血肿周围神经元凋亡细胞:模型组于术后6h开始增多,为(31.24±4.55)个/高倍视野。术后1d明显增多,为(72.58±7.23)个/高倍视野,术后3d达高峰[(159.42±12.26)个/高倍视野],术后5d减少[(90.98±7.65)个/视野];模型组术后各时间点明显多于假手术组[(1.73±0.37),(1.45±0.30),(1.43±0.32),(1.41±0.13)个/高倍视野,P<0.01]。黄芪多糖组术后1,3,5d神经元凋亡细胞为(57.64±8.45),(132.65±8.48),(61.23±7.12)个/高倍视野,少于模型组(P<0.05～0.01)。②血肿周围核因子κB阳性细胞数:模型组与治疗组术后6h在血肿周围及皮质部位见大量核因子κBp65阳性细胞,术后3d达高峰,术后6h～5d各时间点均明显高于假手术组(P<0.01);治疗组术后6h,1,3,5d为(19.21±3.87),(44.28±5.76),(53.29±5.67),(27.82±2.74)个/高倍视野,均少于模型组[(26.75±4.08),(58.33±8.13),(68.15±6.89),(37.98±4.57)个/高倍视野,P<0.05～0.01]。③神经元超微结构:假手术组基本正常;治疗组神经元细胞损伤程度轻于模型组。结论:黄芪多糖可能是通过抑制核因子κB表达来减轻脑出血后的细胞凋亡,对脑出血损伤有神经保护作用。
AIM： To investigate the effects of astragalus polysaccharide on the apnptnsis of nerve cells and expression of nuelear factor （NF）-κB protein in rats after experimental intracerebral hemorrhage（ICH）. METHODS： The experiment was conducted at the Central Laboratory of Nanjing Medical University from January to June 2006. ①Sixty-nine healthy male SD rats were randomly divided into sham-operation group （n=23）, model group （n=23） and treatment group （n=23）; The latter two groups were observed in 4 different time points respectively, such as 6 hours, 1 day, 3 clays and 5 clays after ICH. Three rats in each group were used in the electron microscope test 3 days after operation. ②ICH model was established in model group and treatment group by injection of cnllagenase Ⅶ. Sham-operation group was established by saline of the same volume instead. Treatment group were injected with 25 mg/kg astragalus pnlysaccharide （prnvided by Pharmagenesis, batch number 8 k01101） by intraperitoneal injection once a clay 2 hours after establishing models. The other two groups received 2 mL saline at the same time point by intraperitoneal injection.③ The dynamic change of apoptosis and the expression of NF-κB in the tissue surrounding hematoma were examined with in situ end-labeling （ISEL） and immunohistochemical method. ④ Change of uhrastructures of the neurons around the hematoma were observed with transmission electron microscope 3 days after operation. RESULTS： A total of sixty-nine rats were involved in the analysis of results. ①The numher of apoptotic cells around hematoma after ICH began to increase at the 6^th hour after operation in the model group （31.24±4.55） per high power field, significantly increased at the first day after operation （72.58±7.23） per high power field, reached its peak at the 3^th day after operation [（159.42±12.26） per high power field, and the number gradually decreased at 5^th day（90.98±7.65） per field. It was more obviously in the model group than sham-operation group at each time point after operation [（ 1.73±0.37 ）,（1.45±0.30）,（1.43±0.32）,（1.41±0.13） per high power field, P 〈 0.01]. The number of apoptntic cells of neurons was （57.64±8.45）, （132.65±8.48）,（61.23±7.12） per high power field in the treatment gruup at days 1, 3, 5 after operation, which was fewer than that in the model group （P 〈 0.05-0.01 ）. ②Number of positive cells of NF-κB around hematoma： A great quantity of NF-κBp65 positive cells appeared around hematnma and at cortical part in the model group and treatment group 6 hours after operation, reached the peak 3 days after operation, and was higher remarkably than that in the sham-operation group from hour 6 to day 5 after operation （P 〈 0.01）. The number of positive cells was （ 19.21 ±3.87 ）,（44.28±5.76）,（53.29±5.67）,（27.82±2.74） per high power field at hour 6, days 1, 3, 5 after operation, which was fewer than that in the model group [（26.75±4.08）,（58.33±8.13）,（68.15±6.89）,（37.98±4.57） per high power field,P 〈 0.05-0.01]. ③Ultrastructure of neurons： It was normal in the sham-operation group; the injured level of neurons in the treatment group was lighter than that in the model group. CONCLUSION： Astragalus polysaceharide has a neuroprotective effect on ICH by down-regulating the expression of NF-κB protein and inhibiting the apoptosis of nerve cells.
出处
《中国临床康复》
CSCD
北大核心
2006年第35期13-15,F0003,共4页
Chinese Journal of Clinical Rehabilitation
