蜂胶水提液影响血管紧张素Ⅱ促血管平滑肌细胞增殖的作用  

Influence of water extract of propolis on proliferation of vascular smooth muscle cell induced by angiotensin Ⅱ

作　　者：桑慧[1] 王家富[1] 商战平[1] 李卫红[1] 姚树桐[1] 司艳红[1] 

机构地区：[1]泰山医学院病理生理教研室,山东省泰山市271000

出　　处：《中国临床康复》2006年第35期57-59,共3页Chinese Journal of Clinical Rehabilitation

基　　金：山东省自然基金课题(Z2002C40)~~

摘　　要：目的:观察蜂胶水提液对血管紧张素Ⅱ诱导的人脐动脉平滑肌细胞增殖作用的影响。方法:实验于2005-03/2006-04在泰山医学院基础医学研究所完成。组织块法培养人脐动脉平滑肌细胞,取3～5代细胞进行实验。将培养的人脐动脉平滑肌细胞随机分为4组:①对照组:不加任何药物。②模型组:加入100nmol/L的血管紧张素Ⅱ。③蜂胶组:分别加入50mg/L、100mg/L、200mg/L的蜂胶水提液预处理2h后,再加入终浓度为100nmol/L的血管紧张素Ⅱ。④氯沙坦组:加入1.5μmol/L的氯沙坦预处理2h后,再加入终浓度为100nmol/L的血管紧张素Ⅱ。同步化后,按分组分别加入处理药物孵育24h,倒置显微镜下动态观察细胞形态学的变化,同时计数细胞存活率,并用血细胞计数板计数各组细胞。采用流式细胞仪检测细胞周期,并分析细胞不同周期所占比例。采用免疫细胞化学检测增殖细胞核抗原。计算增殖细胞核抗原阳性细胞百分率。增殖细胞核抗原阳性细胞百分率(%)=增殖细胞核抗原阳性细胞总数/血管平滑肌细胞总数×100%。结果:①相差倒置显微镜下观察各组细胞加药前后形态学无明显变化,细胞存活率均达95.0%以上。②模型组细胞计数高于对照组和氯沙坦组(P<0.01),100mg/L和200mg/L蜂胶组低于模型组(P<0.01);50mg/L和100mg/L蜂胶组高于氯沙坦组(P<0.01),200mg/L蜂胶组与氯沙坦组差异无显著性(P>0.05)。③模型组G0/Gl期细胞百分比低于对照组、氯沙坦组(P<0.01);100mg/L和200mg/L蜂胶组高于模型组(P<0.01);50mg/L蜂胶组低于氯沙坦组(P<0.05),100mg/L、200mg/L蜂胶组与氯沙坦组差异显著性(P>0.05)。④模型组增殖细胞核抗原阳性率高于对照组、氯沙坦组(P<0.01);100mg/L和200mg/L蜂胶组低于模型组(P<0.01),蜂胶各浓度组与氯沙坦组间差异无显著性意义。结论:血管紧张素Ⅱ诱导血管平滑肌细胞的增殖,氯沙坦能阻滞该作用;一定浓度的蜂胶水提液AIM： To observe the effect of water extract of propolis （WEP） on proliferation of human umbilical vascular smooth muscle cell proliferation （HUVSMC） induced by angiotensin Ⅱ （Ang Ⅱ）＋ METHODS： The experiment was conducted in the Institute of Basic Medical, Taishan Medical College between March 2005 and April 2006. Primary culture of HUASMC was performed using the primary-explant method, and 3-5 passage cells were chosen in experiment. Cultured HU- VSMCs were randomly divided into 4 groups： ①control group：without any drug, ②model group： with 100 nmol/L Ang Ⅱ ; ③propolis group： pretreated with 50 mg/L, 100 mg/L and 200 mg/L WEP for 2 hours on HUASMC, respectively ,then incubated with 100 nmol/L Ang Ⅱ .④Losartan group： pretreated with 1.5 μmol/L lostan for 2 hours, then incubated with 100 nmol/L Ang Ⅱ. Growth arrested, synchronized cells were treated with different drug according to group for 24 hours. Cell dynamic morphological change was observed with invert microscope, cell livability was counted, so number in each group was assessed by blood cell counting board. Cell cycle was tested and cell percent in different cycle was analyzed with flow cytometry. Proliferating cell nuclear antigen （PCNA） was detected with immunocytochemistry staining. PCNA positive cell percent was calculated. PCNA positive cell percent （%） was equal to PCNA positive cell number/VSMC total number ×100%. RESULTS： ①Cell morphology did not change after adding drug under phase contrast invert microscope and cell livability was over 95%. ②Compared with control group and losartan group, the number of model group was higher （P 〈 0.01 ）; Compared with model group, the numbers of 1190 mg/L and 200 mg/L propolis group were lower（P 〈 0.01 ）; Compared to losartan group, the numbers of 50 mg/L, and 100 mg/L propolis groups were higher（P 〈 0.01 ）; There was no significant difference of cell counting between 200 mg/L propolis group and losartan group （P 〉 0.05 ）. ③Comp

关 键 词：蜂胶 肌细胞 平滑肌 血管紧张素Ⅱ 细胞计数 细胞分裂期 

分 类 号：R282.71[医药卫生—中药学]