摘要
目的 探讨血管平滑肌细胞(SMC)对光敏剂血啉甲醚(HMME)的吸收特点及影响因素,为进一步建立动脉粥样硬化和血管成形术后再狭窄的光动力治疗方法提供实验依据。方法 培养兔主动脉SMC并接种至96孔培养板。首先,培养液中分别加入HMME和血卟啉衍生物(HpD),浓度分别为20、40、60、80、100μgml,孵育时间分别为0、15、30、60、120、180、240min,采用荧光分析法测定细胞内光敏剂含量,绘制SMC吸收光敏剂的浓度含量和时间含量关系曲线。其次,将SMC分成血小板生长因子组、胎牛血清组和对照组,前两组培养液中分别加入10ngml血小板生长因子或10%胎牛血清,12h后进行光敏剂含量测定、细胞计数,并用MTT比色法测定琥珀酸脱氢酶活性。结果 SMC对HMME和HpD的吸收量随孵育浓度的增高和孵育时间的延长而增加,但HMME的曲线上升斜率明显大于HpD。SMC对HMME的吸收量可较HpD高130%～170%(P<005)。血小板生长因子或胎牛血清作用12h后,两组SMC数量与对照组比较差异均无显著性(P>005),但SMC琥珀酸脱氢酶活性血小板生长因子组较对照组增加621%(P<005),胎牛血清组较对照组增加1194%(P<005);同时,SMC吸收HMME的浓度含量和时间含量关系曲线均明显上移,其关系为胎牛血清组>血小板生长因子组>对照组(P<005)。
Objective To investigate the absorption characteristics of hematoporphyrin monomethyl ether (HMME) by vascular smooth muscle cells (SMC) and to provide a basis for suitable photodynamic therapy (PDT) for atherosclerosis and restenosis. Methods The SMC from rabbit normal abdominal aorta were cultured and divided into three groups. First, the absorption characteristics of HMME and hematoporphyrin derivative(HpD) in several concentrations and for different incubation time in SMC were investigated by using fluorospectrophotometer. Secondly, SMC were incubated in serum free medium, 10 ng/ml PDGF BB or 10% fetal bovine serum (FBS) respectively for 12 hours, and the activity status of SMC, cell number and content of HMME in SMC were investigated. Results The absorption curves of HMME and HpD by SMC were similar, but the content of HMME in SMC was significantly higher than that of HpD (P<0 05). The SMC number did not increase obviously after incubation with growth factors (P>0 05), but the activity status of SMC was 62 1% higher in PDGF group and 119 4% higher in FBS group than that in control group(P<0 05). The curves of HMME uptake in PDGF group and in FBS group ascended significantly (P<0 05).Conclusions The content of HMME in SMC is dose dependent and time dependent, and is significantly higher than that of HpD. The HMME uptake is significantly affected by the metabolic status of SMC.
出处
《中国激光医学杂志》
CAS
CSCD
2000年第1期1-5,共5页
Chinese Journal of Laser Medicine & Surgery
基金
军队"九五"杰出人才基金!98J0 12
关键词
血管平滑肌细胞
血淋甲醚
血卟啉衍生物
Vascular smooth muscle cells
Hematoporphyrin monomethyl ether
Hematoporphyrin derivative
