提睾肌再灌注损伤模型碱性成纤维细胞生长因子变化与丹参的干预效应

Effect of Danshen on basic fibroblast growth factor level in the rat cremaster muscle during ischemia/reperfusion

目的:观察丹参对缺血再灌注损伤提睾肌模型碱性成纤维细胞生长因子变化的影响,以及丹参干预后血清中超氧化物歧化酶活性、丙二醛浓度的变化。方法:实验于2006-02/09在福建中医学院骨伤系创伤修复与重建实验室及福建师大生命科学院实验室完成。实验分组:选用清洁级雄性SD大鼠84只,手术造成左提睾肌缺血再灌注损伤模型,按随机数字表法分为丹参组(n=42)与生理盐水组(n=42),缺血再灌注前30min腹腔分别注射相同剂量的丹参注射液(正大青春宝制药厂,主要成分:丹参,含生药1.5g/mL,批号:0408113,给药量按人剂量与大鼠剂量之间的换算方法,1.809g/kg)、生理盐水。分别于缺血再灌注后10,20,30,40,50,60,90min时抽取腹主动脉血,每个时相点6只。实验评估:①采用考马斯亮兰蛋白法和酶链免疫吸附法测定提睾肌碱性成纤维细胞生长因子含量。②采用黄嘌呤氧化酶法测定血清中超氧化物歧化酶活性。③采用硫代巴比妥酸法测定丙二醛浓度。结果:纳入大鼠84只,均进入结果分析。①提睾肌碱性成纤维细胞生长因子含量:再灌注10,20min丹参组与生理盐水组无明显差别,再灌注30,40,50,60,90min丹参组单位蛋白中碱性成纤维细胞生长因子含量高于生理盐水组,差异有显著性意义(t=2.415,2.292,4.605,3.595,5.196,P<0.05,0.01)。②血清超氧化物歧化酶活性:再灌注开始10min时,丹参组与生理盐水组超氧化物歧化酶活性无明显差异;再灌注20,30,40,50,60,90min丹参组超氧化物歧化酶活性高于生理盐水组,差异均有显著性意义(t=2.949,2.327,4.050,2.958,9.142,6.188,P<0.05,0.01)。③血清丙二醛浓度:再灌注10,20min时,丹参组与生理盐水组丙二醛浓度无明显差异,再灌注30,40,50,60,90min丹参组丙二醛浓度低于生理盐水组,差异有显著性意义(t=3.164,2.697,4.461,2.587,3.002,P<0.05,0.01)。结论:丹参能促进再灌注损伤中骨骼肌碱性成纤维细胞生长因子的表达,减少骨骼肌缺血再灌注损伤过程中产生的丙二醛并促进血清中超氧化物歧化酶的分泌,这可能亦是丹参防治缺血再灌注损伤的机制之一。

AIM： To observe the effect of Danshen on basic fibroblast growth factor （bFGF）, serum superoxide dismutase （SOD）, and malondialdehyde （MDA） in rat models of ischemia/reperfusion （I/R）. METHODS： The experiment was conducted in the laboratory of College of Life Science, Fujian Normal University and Department of Orthopaedics and Traumatology, Fujian University of Traditional Chinese Medicine from February to September 2006.①The I/R models of cremaster muscles were established in 84 male SD rats of clean grade. Then the rats were randomly divided into Danshen group （n =42） and normal saline group （n =42）, which were intraperitoneally injected with Danshen （No. 0408113, Chiatai Qingchunbao Pharmaceutical, Co., Ltd. composed of Danshen, 1.5 g/mL crude drug, the administration dose was accorded with the conversion between human and rats, 1.809 g/kg） or normal saline 30 minutes before reperfusion. Blood of abdominal aorta was drawn after reperfusion for 10, 20, 30, 40, 50, 60, and 90 minutes, respectively, with 6 rats at each time point. ②Coomassie brilliant blue and ELISA methods were adopted to determine the content of bFGF; SOD activity and MDA levels in serum were detected by xanthine oxidase and thiobarbituric acid methods, respectively. RESULTS： Totally 84 rats were involved in the result analysis. ①There was no obvious difference in bFGF content between two groups after reperfusion for 10 and 20 minutes, but the content in Danshen group was significantly increased compared with normal saline group after reperfusion for 30, 40, 50, 60 and 90 minutes （t =2.415, 2.292, 4.605, 3.595, 5.196, P〈 0.05, 0.01）. ②No distinct difference was found in serum SOD activity between two groups after reperfusion for 10 minutes, but the activity in Danshen group was significantly increased compared with normal saline group after reperfusion for 20, 30, 40, 50, 60 and 90 minutes （t =2.949, 2.327, 4.050, 2.958, 9.142, 6.188, P 〈 0.05, 0.01）. ③There was no obvious difference in MDA content between two groups after reperfusion for 10 and 20 minutes, but the content in Danshen group was significantly increased compared with normal saline group after reperfusion for 30, 40, 50, 60 and 90 minutes （t =3.164, 2.697, 4.461, 2.587, 3.002, P〈 0.05, 0.01）. CONCLUSION： Danshen could promote the expression of bFGF and production of SOD but reduce the content of MDA in cremaster muscles of rats with I/R injury. This may be one of the mechanisms of Danshen to prevent I/R injury.