植物药有效成分β-蜕皮激素抑制血管内皮细胞的凋亡

Prohibitive effects of beta-ecdysone on apoptosis of vascular endothelial cells

目的:分析昆虫变态激素β-蜕皮激素是否能抑制血管内皮细胞的凋亡。方法:实验于2003-09/2006-03在南方医科大学病理生理实验室完成。人内皮细胞株ECV-304消化后制成单细胞悬液,接种至无菌培养管(2mL/管)。分为3组:①空白对照组。②模型组:包括40,80,160mmol/L亚砷酸钠3个剂量组。③实验组:包括160mmol/L亚砷酸钠+50,200,800mg/Lβ-蜕皮激素3个剂量组。培养24h后测定相关指标,应用流式细胞术测定内皮细胞的凋亡细胞数及平均通道荧光值,取平均通道荧光值对数,对数值越大,氧化水平越高。结果:①40,80,160mmol/L亚砷酸钠模型组的血管内皮细胞凋亡率均显著高于对照组(16.70±3.56)%,(26.80±2.48)%,(54.30±11.06)%,(7.67±2.42)%(P<0.01)。②50mg/L和200mg/Lβ-蜕皮激素实验组的血管内皮细胞凋亡率均显著低于160mmol/L亚砷酸钠模型组(P<0.01),但800mg/Lβ-蜕皮激素实验组显著高于160mmol/L亚砷酸钠模型组(P<0.05)。③40,80,160mmol/L亚砷酸钠模型组血管内皮细胞的氧化水平均显著低于对照组(P<0.01),50,200mg/Lβ-蜕皮激素实验组能使氧化水平有所升高,800mg/Lβ-蜕皮激素实验组能使氧化水平有所降低,但与亚砷酸钠模型组相比,差异无显著性意义穴P>0.05雪。结论:亚砷酸钠可剂量依赖地诱导内皮细胞凋亡;β-蜕皮激素能影响亚砷酸钠诱导的内皮细胞凋亡,但具体剂量关系值得进一步分析。

AIM： To analyze whether insect moulting hormone β-ocdysone can inhibit the apoptosis of vascular endothelial cells （VECs）. METHODS： The experiment was conducted at the Laboratory of Pathophysiology, Southern Medical University between September 2003 and March 2006. Human EC line ECV-304 after digestion was used to prepare monocellular suspension, and then inoculated into sterile culture tube （2 mL a tube）. Three groups were designed： ①contrul gronp ②model group： throe subgroups of sodium arsenite （Ars） at 40, 80 and 160 mmol/L ③experimental group： throe subgroups of 160 mmol/L Ars＋50, 200, 800 mg/L β- ecdysone. Flow cytometry was applied to detect the amount of apoptotic ECs and mean fluorescent value, whose logarithm was also obtained. The greater logarithm indicated the higher oxidative level. RESULTS： ①The apoptosis rate of VECs in model groups of 40, 80 and 160 mmol/L Ars were remarkably higher than that of control group [（16.70±3.56）%, （26.80±2.48）%, （54.30±11.06）%, （7.67±2.42）%, P 〈 0.01]. ②Comparod with 50 mg/L and 200 mg/L β-ecdysone, 160 mmol/L Ars exhibited the stronger effect for inhibiting apoptosis （P 〈 0.01）. However, 800 mg/L β-ecdysone increased significantly the amount of apoptosis cells than 160 mmol/L Ars （P 〈 0.05）.③The oxidative levels of ECs were significantly lower in three Ars model groups than in control group （P 〈 0.01）. In addition, 50 mg/L and 200 mg/L β-ecdysone elevated the oxidative level whereas 800 mg/L β-ecdysone reduced oxidative level, with the insignificant difference compared with Ars model group （P 〉 0.05）. CONCLUSION： Ars can induce apoptosis of ECs dosage-dependently; β- ecdysone may influence apoptosis of ECs induced by Ars. Further analysis need to reveal the dose-dependence.