萝卜硫素对内皮素-1诱导大鼠平滑肌细胞增殖的影响  

Effects of sulforaphane on endothelin-1-induced proliferation in rat vascular smooth muscle cells

作　　者：汪晨净[1] 裴淑艳[1] 王晓坤 孙枫 郭忠[1] WANG Chen-jing;PEI Shu-yan;WANG Xiao-kun;SUN Feng;GUO Zhong(Department of Pharmacology,School of Basic Medical Sciences,Northwest Minzu University Health Science Center,Lanzhou 730030,Gansu Province,China)

机构地区：[1]西北民族大学医学部基础医学院药理学教研室,甘肃兰州730030

出　　处：《中国临床药理学杂志》2024年第3期363-367,共5页The Chinese Journal of Clinical Pharmacology

基　　金：中央高校基本科研业务费基金资助项目(31920230053);兰州市科技计划医疗卫生专项基金资助项目(2021-1-133);甘肃省自然科学基金资助项目(20JR5RA503)。

摘　　要：目的 研究萝卜硫素(SFN)对内皮素-1(ET-1)诱导大鼠胸主动脉血管平滑肌细胞(VSMC)增殖的影响及其机制。方法 将组织贴块法原代培养的VSMC随机分为对照组(正常培养)、模型组(给予0.1μmol·L^(-1 )ET-1处理)和低、中、高剂量实验组(分别给予5、10、20μmol·L^(-1 )SFN+0.1μmol·L^(-1) ET-1处理)。用噻唑蓝法检测细胞增殖活力,用实时荧光定量聚合酶链反应法测定增殖及凋亡相关基因mRNA表达水平,用荧光探针2’,7’-二氯二氢荧光素二乙酸酯法检测活性氧(ROS)相对荧光强度,用蛋白质印迹法测定B淋巴细胞瘤-2(Bcl-2)、半胱氨酸天冬氨酸蛋白酶-3(caspase-3)、p38丝裂原活化蛋白激酶(p38MAPK)、磷酸化p38丝裂原活化蛋白激酶(p-p38MAPK)及p53蛋白的表达水平。结果 对照组、模型组和低、中、高剂量实验组的细胞增殖活力分别为0.27±0.02、0.50±0.04、0.40±0.03、0.38±0.03和0.34±0.03,Bcl-2 mRNA表达水平分别为1.57±0.28、1.00±0.00、0.87±0.05、0.75±0.08和0.41±0.18,caspase-3 mRNA分别为0.84±0.10、1.00±0.00、1.33±0.10、1.61±0.15和1.83±0.16,ROS相对荧光强度分别为6.05±1.50、31.45±3.12、30.03±1.85、18.39±5.62和17.18±1.97,Bcl-2蛋白相对表达水平分别为0.65±0.02、0.60±0.02、0.49±0.02、0.48±0.03和0.49±0.01,caspase-3蛋白相对表达水平分别为0.03±0.00、0.25±0.01、0.42±0.01、0.46±0.02和0.64±0.03,p-p38MAPK/p38MAPK蛋白相对表达水平比值分别为0.97±0.05、1.44±0.04、1.62±0.10、2.18±0.05和2.70±0.05,p53蛋白相对表达水平分别为0.20±0.01、0.30±0.01、0.34±0.02、0.37±0.01和0.42±0.01。模型组的上述指标与对照组比较,中、高剂量实验组的上述指标与模型组比较,在统计学上差异均有统计学意义(P<0.05,P<0.01)。结论 SFN可抑制ET-1诱导的VSMC增殖,促进VSMC凋亡,其机制可能与SFN抗氧化、激活p38MAPK及p53依赖的细胞凋亡途径有关。Objective To investigate the effects and the mechanisms of sulforaphane (SFN) on endothelin-1 (ET-1)-induced proliferation in rat vascular smooth muscle cell (VSMC).Methods Rat VSMC were cultured by using the modified tissue explant technique in vitro and were randomly divided into control group (normal culture),model group(treated with 0.1μmol·L^(-1)ET-1) and experimental-L,-M,-H groups (treated with 5,10,20 SFN+0.1μmol·L^(-1)ET-1).The survival rate of VSMC were measured by methyl thiazolyl tetrazolium.The proliferation and apoptosis related genes in mRNA level in VSMC were respectively examined with real-time quantitative polymerase chain reaction;the relative fluorescence intensity of reactive oxygen species (ROS) in VSMC were measured by fluorescent probe 2’,7’-dichlorodihydrofluorescein diacetate;the protein expression of B-cell lymphoma-2 (Bcl-2),cysteinyl aspartate-specific proteinase-3 (caspase-3),p38 mitogen-activated protein kinases (p38MAPK),phosphorylated p38 mitogen-activated protein kinases (p-p38MAPK),and p53 in VSMC were investigated by Western blot.Results The proliferation activities of the control group,model group,experimental-L,-M,-H groups were 0.27±0.02,0.50±0.04,0.40±0.03,0.38±0.03 and 0.34±0.03;the expression levels of Bcl-2 mRNA were 1.57±0.28,1.00±0.00,0.87±0.05,0.75±0.08 and 0.41±0.18;the expression levels of caspase-3 mRNA were 0.84±0.10,1.00±0.00,1.33±0.10,1.61±0.15 and1.83±0.16;the relative fluorescence intensities of ROS in VSMC were 6.05±1.50,31.45±3.12,30.03±1.85,18.39±5.62 and 17.18±1.97;the relative expression levels of Bcl-2 protein were 0.65±0.02,0.60±0.02,0.49±0.02,0.48±0.03 and 0.49±0.01;the relative expression levels of caspase-3 protein were 0.03±0.00,0.25±0.01,0.42±0.01,0.46±0.02 and 0.64±0.03;the protein expression ratios of p-p38MAPK/p38MAPK were 0.97±0.05,1.44±0.04,1.62±0.10,2.18±0.05 and 2.70±0.05;the relative expression levels of p53protein were 0.20±0.01,0.30±0.01,0.34±0.02,0.37±0.01 and 0.42±0.01 respectively.There w

关 键 词：萝卜硫素 增殖 内皮素-1 凋亡 血管平滑肌细胞 

分 类 号：R28[医药卫生—中药学]