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本室在实验性动脉粥样硬化研究过程中,建立了体外培养动脉壁平滑肌细胞(SMC)高脂损伤模型。为了观察SMC中粘多糖的存在及性质及其与动脉粥样硬化病变的关系,采用阿新蓝一钌红法,显示SMC内羟基酸性粘多糖和硫酸基粘多糖。一材料 1.标本制备:取新小牛主动脉平滑肌。(1)在无菌条件下,剔除动脉内外膜,将中膜剪成小块,置于胶原酶、弹性蛋白酶1∶1混合液中,在37℃条件下进行孵育,边孵边搅动,孵育6小时后,弃除溶液中胶原组织,900转/分离心5分钟。Hank’s液清洗沉淀物三次,然后加入含小牛血清(cs)的IMDM培养液,在37℃封闭培养,每隔四日换液一次,分别培养15天,72天。(2)刮下SMC,用蒸馏
Our laboratory established experimental models of atherosclerosis in vitro cultured rat arterial wall smooth muscle cells (SMC) model of high-fat injury. In order to observe the presence and nature of mucopolysaccharides in SMC and its relationship with atherosclerotic lesions, amoxicillin-ruthenium red method was used to reveal the presence of hydroxy acid mucopolysaccharides and sulpham mucopolysaccharides in SMCs. A material 1. Specimen preparation: Take new cows aorta smooth muscle. (1) under aseptic conditions, remove the inner and outer arterial membrane, the membrane cut into small pieces, placed collagenase, elastase 1: 1 mixture, incubated at 37 ℃ conditions, stirring while incubation, incubation After 6 hours, the collagen solution was discarded and centrifuged at 900 rpm for 5 minutes. Hank’s solution was washed three times with sediment, and then IMDM medium containing bovine serum (cs) was added. The medium was blocked at 37 ° C and changed every four days for 15 days and 72 days respectively. (2) scraped SMC, with distillation