摘要
目的探讨地黄多糖(RPS)对血管钙化(VC)的作用及其作用机制。方法将8周龄雄性SD大鼠按随机数目表法分配原则分为3组:正常对照组(Con)、VC、VC+RPS组,每组12只。采用维生素D3(VD3)和尼古丁制备大鼠VC模型。VC+RPS组在造模后第2d开始予RPS灌胃,Con和VC组每天给予同剂量生理盐水灌胃,4周后处死动物并取材。茜素红染色和原位缺口末端标记法(TUNEL)检测VC及凋亡;试剂盒检测血管中碱性磷酸酶(ALP)和钙离子含量;蛋白印迹法(Western blot)检测主动脉中Runx相关转录因子2(Runx2)、α-肌动蛋白(α-SMA)、B淋巴细胞瘤因子相关X蛋白(bax)、B淋巴细胞瘤2(bcl-2)、活化型半胱氨酸天冬氨酸蛋白酶3(cleaved Caspase-3)蛋白表达水平。多组间均数比较采用单因素方差分析。结果VC组ALP活性及钙含量高于Con组[(219.80±32.70)U/g、(197.32±28.30)μmol/g比(53.20±12.20)U/g、(20.46±5.30)μmol/g,F=202.800、247.000,P<0.01],VC+RPS组ALP活性及钙含量低于VC组[(122.00±21.50)U/g、(78.45±12.60)μmol/g比(219.80±32.70)U/g、(197.32±28.30)μmol/g,q=16.630、20.720,P<0.01];Tunel染色结果显示,VC组凋亡指数明显高于Con组[(62.30±7.40)%比(1.30±0.14)%,F=197.700,P<0.01],VC+RPS组凋亡指数低于VC组[(45.70±6.20)%比(62.30±7.40)%,q=7.394,P<0.01]。VC组中α-SMA蛋白相对表达量低于Con组(0.74±0.03比1.00±0.03,F=26.870,P<0.01),VC+RPS组中α-SMA蛋白相对表达量高于VC组(1.01±0.25比0.74±0.03,q=9.037,P<0.01);VC组中Runx2表达高于Con组(1.75±0.08比1.00±0.05,F=28.250,P<0.01),VC+RPS组中Runx2表达低于VC组(1.44±0.07比1.75±0.08,F=35.241,P<0.05);VC组中bax蛋白相对表达量高于Con组(2.65±0.21比1.00±0.04,F=113.400,P<0.01),VC+RPS组中bax蛋白表达量低于VC组(1.51±0.12比2.65±0.21,q=6.375,P<0.01);VC组中bcl-2蛋白相对表达量低于Con组(0.51±0.05比1.00±0.02,F=64.280,P<0.01),VC+RPS组中bcl-2蛋白表达高于VC组(0.79±0.01比0.51±0.05,q=9.061,P<0.01);VC组中Cleaved Caspase-3蛋白相对表�
Objective To investigate the effect of rehmanniaglutinosa polysaccharide(RPS)on vascular calcification(VC)and its mechanisms.Methods Male Sprague Dawley rats aged 8 weeks were randomly divided into 3 groups:control group(Con),VC group and VC+RPS group,with 12 rats in each group.The rat model of VC was established by injection of vitamin D3 and nicotine.The VC+RPS group was given an additional treatment of RPS on the second day,while Con and VC groups were given the same dosage of normal saline every day.Four weeks later,the animals were sacrificed and the aorta was peeled off.VC and cell apoptosis were individually detected by alizarin red staining and terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)staining.The alkaline phosphatase(ALP)and calcium ion in vascular tissue were determined according to the kit protocol.The expression levels of Runx-related transcription factor 2(Runx2),alpha smooth muscle actin(α-SMA),B-cell lymphoma 2 associated X protein(bax),B-cell lymphoma 2(bcl-2)and cleaved Caspase-3 in aorta were detected by Western blotting.The means of multiple groups were compared by single factor analysis of variance.Results The ALP activity and calcium content in VC group were higher than Con group[(219.80±32.70)U/g,(197.32±28.30)μmol/g vs.(53.20±12.20)U/g,(20.46±5.30)μmol/g,F=202.800,247.000,P<0.01],VC+RPS group were lower than VC group[(122.00±21.50)U/g,(78.45±12.60)μmol/g vs.(219.80±32.70)U/g,(197.32±28.30)μmol/g,q=16.630,20.720,P<0.01]Tunel staining showed that the apoptosis index in VC group was significantly higher than Con group[(62.30±7.40)% vs.(1.30±0.14)%,F=197.700,P<0.01],the VC+RPS group was lower than VC group[(45.70±6.20)% vs.(62.30±7.40)%,q=7.394,P<0.01].The expression ofα-SMA in VC group were lower than Con group(0.74±0.03 vs.1.00±0.03,F=26.870,P<0.01),the VC+RPS group was higher than VC group(1.01±0.25 vs.0.74±0.03,q=9.037,P<0.01).The expression of Runx2 in VC group were higher than Con group(1.75±0.08 vs.1.00±0.05,F=28.250,P<0.01),the VC+RP
作者
高帅
泮思林
杜占慧
罗刚
纪志娴
Gao Shuai;Pan Silin;Du Zhanhui;Luo Gang;Ji Zhixian(Heart Center,Qingdao Women and Children's Hospital,Qingdao University,Qingdao 266034,China)
出处
《中华实验外科杂志》
CAS
北大核心
2021年第8期1466-1469,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(81770316、81970249)
山东省泰山学者工程资助项目(2018)。
关键词
地黄多糖
血管钙化
凋亡
Rehmanniaglutinosa polysaccharide
Vascular calcification
Apoptosis
