盐霉素对大鼠微血管内皮细胞血管生成的抑制  

Salthromycin inhibits angiogenesis of rat microvascular endothelial cells

作　　者：王薇[1] 王俊壹 薛兴功 宋玮[1] 段秋莉 肖荣[1] 张普[1] Wang Wei;Wang Junyi;Xue Xinggong;Song Wei;Duan Qiuli;Xiao Rong;Zhang Pu(Department of Intensive Care Unit,Taian City Central Hospital,Taian 271000,China;Department of Cardiovascular Medicine,Shengzhuang Village Hospital of Taishan District,Taian 271039,China;Neonatal Department,Taian Maternal and Child Healthcare Hospital,Taian 271000,China)

机构地区：[1]泰安市中心医院重症医学科,山东泰安271000 [2]泰安市泰山区省庄卫生院心内科,山东泰安271039 [3]泰安市妇幼保健院新生儿科,山东泰安271000

出　　处：《中华临床医师杂志（电子版）》2019年第4期290-293,共4页Chinese Journal of Clinicians(Electronic Edition)

摘　　要：目的探讨盐霉素是否可通过抑制大鼠微血管内皮细胞的增殖、迁移、侵袭抑制血管生成。方法取6~8周雄性Wistar大鼠心脏,应用植块法原代培养大鼠微血管内皮细胞,取第三代细胞进行实验。给予不同浓度的盐霉素刺激大鼠微血管内皮细胞,采用MTT方法检测盐霉素对大鼠微血管内皮细胞增值能力的影响。采用细胞损伤划痕实验检测盐霉素对大鼠微血管内皮细胞迁移能力的影响,应用Transwell实验方法检测盐霉素对大鼠微血管内皮细胞侵袭能力的影响。采用体外毛细血管管腔样结构形成方法观察盐霉素对毛细血管管腔样结构官腔形成能力的影响,验证盐霉素对大鼠微血管内皮细胞血管生成能力的影响。采用t检验比较对照组与各实验组之间吸光度值、划痕距离差、Transwell细胞数以及体外毛细血管管腔样结构形成条数的差异。结果MTT检测细胞增殖能力显示,对照组吸光度值(0.968±0.038)相比SAL在1μM(0.849±0.031)时已高,且差异具有统计学意义(t=4.11,P=0.0367),即可明显抑制细胞增殖,且呈浓度依赖性。细胞划痕损伤修复能力检测结果显示,与对照组划痕距离[(249.78±6.72)μm]相比,VEGF组划痕距离[(289.67±8.59)μm]更大,促进了划痕修复,差异具有统计学意义(t=4.76,P=0.0165),SAL组划痕距离[(98.59±4.78)μm]减小,抑制了划痕修复,差异具有统计学意义(t=30.6,P=0.0007)。Transwell实验结果显示,与对照组细胞数(128.67±4.89)个相比,VEGF组细胞数[(158.23±5.26)个]增加,促进了细胞侵袭,差异具有统计学意义(t=5.65,P=0.0089),SAL组细胞数[(98.59±4.78)个]减少,抑制了细胞侵袭,差异具有统计学意义(t=25.65,P<0.001)。体外毛细血管管腔样结构形成实验结果显示,与对照组[(31.29±1.68)条]比较,VEGF组形成[(45.76±2.56)条]更多,促进了血管新生,差异具有统计学意义(t=3.90,P=0.0176),SAL组形成[(14.38±1.23)条]减少,抑制了血管新生,差异具Objective To investigate whether salinomycin (SAL) inhibits angiogenesis by inhibiting the proliferation, migration, and invasion of rat microvascular endothelial cells (MVECs). Methods Male Wistar rats (6-8 weeks old) were used in this study. Primary rat MVECs were cultured and cells at passage 3 were used for subsequent experiments. MVECs were stimulated with different concentrations of SAL in vitro. MTT method was used to detect the proliferation of MVECs. Scratch and Transwell assays were used to detect the migration and invasion of HVECs. Capillary lumen formation assay was used to verify the effect of SAL on the angiogenesis of MVECs. Statistical analyses were performed using SPSS13. The t-test was used to analyze the difference between different groups. P<0.05 was considered statistically significant. Results MTT assay showed that compared with the control group (0.968±0.038), SAL at a concentration of 1 μM (0.849±0.031) significantly inhibited cell proliferation (t=4.11, P=0.0367), and the inhibition was concentration-dependent. The results of scratch assay showed that compared with the control group [scratch distance,(249.78±6.72)μm], VEGF [(289.67±8.59)μm] significantly promoted scratch repair (t=4.76, P=0.0165), and SAL [(98.59±4.78)μm] significantly inhibited scratch repair (t=30.6, P=0.0007). The results of Transwell assay showed that compared with the radiotherapy group (128.67±4.89), VEGF (158.23±5.26) significantly promoted cell invasion (t=5.65, P=0.0089), and SAL (98.59±4.78) significantly inhibited cell invasion (t=25.65, P<0.001). Capillary lumen formation assay showed that compared with the control group (31.29±1.68), VEGF (45.76±2.56) significantly promoted angiogenesis (t=3.90, P=0.0176), and SAL (14.38±1.23) significantly inhibited angiogenesis (t=5.15, P=0.0046). Conclusion SAL can inhibit the angiogenesis of MVECs by inhibiting the proliferation, migration and invasion of rat MVECs.

关 键 词：盐霉素 微血管内皮细胞 迁移 侵袭 血管生成 

分 类 号：R96[医药卫生—药理学]