摘要：
背景:珍珠有镇静安神的药效,临床可应用于高血压病,但水解珍珠液对高血压病的防治作用及相关机制研究目前报道尚少.目的:观察水解南珠液对人微血管内皮细胞生物学行为和分泌功能的影响.方法:人微血管内皮细胞培养,传代.实验分为水解南珠液高、中、低剂量组:分别加入含质量浓度为120, 60,30 mg/L水解南珠液的培养基200 μL;对照组:只加 200 μL的培养基.CCK8法检测人微血管内皮细胞增殖情况,流式细胞术检测细胞的周期分布,TUNEL法了解细胞的凋亡情况,Transwell法检测细胞迁移情况,硝酸还原酶法检测细胞的NO分泌量及化学荧光法检测细胞活性氧的分泌情况.结果与结论:①与对照组相比,低、中、高剂量水解南珠液对人微血管内皮细胞均有促增殖作用,且浓度越高,对细胞的促增殖能力越强(P ＜ 0.05),因此确定120 mg/L水解南珠液为最佳作用浓度;②与对照组相比,水解南珠液组细胞S期和G2期百分比增加,而G1期的细胞比例则减少(P ＜ 0.05),提示水解南珠液可以促进血管内皮细胞的周期进展;③各组均未发现有细胞核被染成绿色的凋亡细胞;④水解南珠液组的迁移细胞数比对照组显著增加(P ＜ 0.05);⑤水解南珠液组细胞NO分泌量明显高于对照组(P ＜ 0.05),活性氧量显著低于对照组(P ＜ 0.05);⑥结果表明,水解南珠液能促进体外培养的人微血管内皮细胞的增殖和迁移,并具有促进其分泌NO及抑制其活性氧分泌的功能,在保护内皮细胞功能方面可能有积极的作用.%BACKGROUND: Pearl has the tranquilizing effect, and it can be applied in the treatment of hypertension. However, there is little report on the prevention and cure effect and mechanism of hydrolyzed pearl liquid on hypertension. OBJECTIVE: To observe the effect of hydrolysis of Hepu pearl (hydrolyzed Nanzhu fluid) on the biological behavior and secretion of human microvascular endothelial cells. METHODS: Human microvascular endothelial cells were cultured and passaged. There were four groups, and the microvascular endothelial cells were incubated in the 200 μL culture medium containing nothing (control group), and 120, 60 and 30 mg/L hydrolysis of Nanzhu fluid. The cell proliferation and migration was detected by cell conuting kit-8 assay and Transwell assay respectively; the cell cycle distribution was tested by flow cytometry; the cell apoptosis was assayed by TUNEL method; the secretion of nitric oxide and reactive oxygen species was tested by nitrale reduetase and chemical fluorescence method, respectively. RESULTS AND CONCLUSION: Compared with the control group, hydrolysis of Nanzhu fluid significantly promoted the proliferation of microvascular endothelial cells in a manner-dependent manner (P ＜ 0.05), suggesting the optimal concentration was 120 mg/L. Compared with the control group, the percentage of cells in S and G2 phase was significantly increased, and the percentage of cells in the G1 phase was significantly reduced in the hydrolysis of Nanzhu fluid group (P＜0.05), indicating hydrolysis of Nanzhu fluid could promote cell cycle progression. Apoptotic cells with green-stained nucleus were invisible in both groups. The number of cell migration in the hydrolysis of Nanzhu fluid group was significantly more than that in the control group (P ＜ 0.05). Compared with the control group, there was a significant increase in the nitric oxide secretion, and a significant decrease in the production of reactive oxygen species in the hydrolysis of Nanzhu fluid group (P＜0.05). To conclude, hydrolysis of Nanzhu fluid can promote the proliferation and migration of human microvascular endothelial cells in vitro, and has the function of promoting the secretion of nitric oxide and inhibiting reactive oxygen species secretion, implying its positive role in the protection of endothelial cell function.
