岭南心血管病杂志
嶺南心血管病雜誌
령남심혈관병잡지
SOUTH CHINA JOURNAL OF CARDIOLOGY
2009年
3期
234-237
,共4页
紫杉醇%肌细胞,平滑肌%血管成形术,经腔,经皮冠状动脉%大鼠
紫杉醇%肌細胞,平滑肌%血管成形術,經腔,經皮冠狀動脈%大鼠
자삼순%기세포,평활기%혈관성형술,경강,경피관상동맥%대서
paclitaxel%smooth muscle cell%percutaneous transcoronary angioplasty%rat
目的 以体外培养的大鼠血管平滑肌细胞为研究对象,探讨紫杉醇对平滑肌细胞增殖的作用.方法 SD大鼠腹腔注射麻醉后,解剖、分离其主动脉,仔细分离血管的平滑肌层并剪成小块置于培养瓶中培养.采用倒置像筹显微镜下观察及α平滑肌肌动蛋白免疫荧光检测对传4代～6代的细胞进行鉴定.根据加入的干预药物将平滑肌细胞分为无血清M199培养基组(A组)、1 nmol/L紫杉醇组(B组)、10 nmol/L紫杉醇组(C组)及100 nmol/L紫杉醇组(D组)共4组,每组均为5个样本.采用流式细胞仪检测不同干预组平滑肌细胞的生长周期,用增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)检测方法 检测细胞的PCNA阳性百分比.结果 培养约2周后出现大量细胞生长,可见致密的细胞层.倒置像差显微镜下观察,细胞呈梭形;随着细胞融合度的增加,可以看到平滑肌细胞特有的"峰-谷"生长现象.培养的血管平滑肌细胞α平滑肌肌动蛋白免疫荧光检测呈阳性,总阳性率大于90%.随着紫杉醇用量的增加,细胞的PCNA阳性百分比呈逐渐减少的趋势,与对照组(A组)比较,10 nmol/L(C组)和100 nmol/L(D组)两种浓度下的紫杉醇引起的PCNA阳性率明显减低.流式细胞仪检测显示,随着紫杉醇用量的增加,细胞S期百分比旱逐渐减少的趋势.与对照组相比,10 nmol/L(C组)和100 nmol/L(D组)两种浓度下的紫杉醇引起的S期百分比明显减低.结论 紫杉醇能够抑制大鼠血管平滑肌细胞的增殖.
目的 以體外培養的大鼠血管平滑肌細胞為研究對象,探討紫杉醇對平滑肌細胞增殖的作用.方法 SD大鼠腹腔註射痳醉後,解剖、分離其主動脈,仔細分離血管的平滑肌層併剪成小塊置于培養瓶中培養.採用倒置像籌顯微鏡下觀察及α平滑肌肌動蛋白免疫熒光檢測對傳4代～6代的細胞進行鑒定.根據加入的榦預藥物將平滑肌細胞分為無血清M199培養基組(A組)、1 nmol/L紫杉醇組(B組)、10 nmol/L紫杉醇組(C組)及100 nmol/L紫杉醇組(D組)共4組,每組均為5箇樣本.採用流式細胞儀檢測不同榦預組平滑肌細胞的生長週期,用增殖細胞覈抗原(proliferating cell nuclear antigen,PCNA)檢測方法 檢測細胞的PCNA暘性百分比.結果 培養約2週後齣現大量細胞生長,可見緻密的細胞層.倒置像差顯微鏡下觀察,細胞呈梭形;隨著細胞融閤度的增加,可以看到平滑肌細胞特有的"峰-穀"生長現象.培養的血管平滑肌細胞α平滑肌肌動蛋白免疫熒光檢測呈暘性,總暘性率大于90%.隨著紫杉醇用量的增加,細胞的PCNA暘性百分比呈逐漸減少的趨勢,與對照組(A組)比較,10 nmol/L(C組)和100 nmol/L(D組)兩種濃度下的紫杉醇引起的PCNA暘性率明顯減低.流式細胞儀檢測顯示,隨著紫杉醇用量的增加,細胞S期百分比旱逐漸減少的趨勢.與對照組相比,10 nmol/L(C組)和100 nmol/L(D組)兩種濃度下的紫杉醇引起的S期百分比明顯減低.結論 紫杉醇能夠抑製大鼠血管平滑肌細胞的增殖.
목적 이체외배양적대서혈관평활기세포위연구대상,탐토자삼순대평활기세포증식적작용.방법 SD대서복강주사마취후,해부、분리기주동맥,자세분리혈관적평활기층병전성소괴치우배양병중배양.채용도치상주현미경하관찰급α평활기기동단백면역형광검측대전4대～6대적세포진행감정.근거가입적간예약물장평활기세포분위무혈청M199배양기조(A조)、1 nmol/L자삼순조(B조)、10 nmol/L자삼순조(C조)급100 nmol/L자삼순조(D조)공4조,매조균위5개양본.채용류식세포의검측불동간예조평활기세포적생장주기,용증식세포핵항원(proliferating cell nuclear antigen,PCNA)검측방법 검측세포적PCNA양성백분비.결과 배양약2주후출현대량세포생장,가견치밀적세포층.도치상차현미경하관찰,세포정사형;수착세포융합도적증가,가이간도평활기세포특유적"봉-곡"생장현상.배양적혈관평활기세포α평활기기동단백면역형광검측정양성,총양성솔대우90%.수착자삼순용량적증가,세포적PCNA양성백분비정축점감소적추세,여대조조(A조)비교,10 nmol/L(C조)화100 nmol/L(D조)량충농도하적자삼순인기적PCNA양성솔명현감저.류식세포의검측현시,수착자삼순용량적증가,세포S기백분비한축점감소적추세.여대조조상비,10 nmol/L(C조)화100 nmol/L(D조)량충농도하적자삼순인기적S기백분비명현감저.결론 자삼순능구억제대서혈관평활기세포적증식.
Objectives To evaluate the inhibitory effect of paclitaxel on proliferation of vascular smooth muscle cell in rats. Methods Isolated the aortae of the rats, drawed off the smooth muscle and cut them into fragment, then put the fragment in the culture flask to cultivate. Verificated the cells by inverted aberratio microscope and α-smooth muscle-actin immunofluorescence detection after they have been passaged for 4-6 times. The smooth muscle cells were divided into 4 different groups according to the adding medicine: group A with non serum M199 medium; group B with 1 nmol/L paclitaxel, group C with 10 nmol/L paclitaxel and group D with 100 nmoi/L paclitaxel. There were 5 samples in each group. Flow cytomery was used to detect the grow periodicity of the smooth muscle cells and vascular smooth muscle cell proliferating cell nuclear antigen (PCNA) were also detected. Results A great quantity of cells growed after 2 weeks. The cells were observed fusiform shape and presented peak-valley grow phenomenon by inverted aberration microscope. The α-smooth muscle-actin immunofluorescence detection of the culture smooth muscle cells were positive and the positive rate was over 90%. The posititve rate of the smooth muscle cell PCNA presented a taper tendency, following the increase dosage of paclitaxel. Compared with group A, the posititve rate of the smooth muscle cell PCNA in group C and group D decreased significantly. The flow cytometry Results indicated that the S period rate of the smooth muscle cell presented a taper tendency, following the increase dosage of paclitaxel. Compared with group A, the S period rate in group C and group D decreased significantly. Conclusions Paclitaxel can inhibit the proliferation of vascular smooth muscle cell in rats.
