血必净注射液对内毒素刺激内皮祖细胞增殖和黏附功能的影响

摘要

目的 观察细菌内毒素脂多糖(LPS )刺激人内皮祖细胞(EPCs )后不同浓度血必净注射液对其增殖、黏附功能的影响.方法 将EPCs 经复苏、培养、传代、鉴定为正在分化的EPCs 后,等量接种到培养板上,随机分为空白对照组[微血管内皮细胞培养液-2(EGM-2MV)]、LPS 组(100 μg/L)、血必净1 组(LPS 刺激EPCs 后加入5、25 、50 g/L 血必净注射液共同孵育)、血必净2 组(LPS 加5、25 、50 g/L 血必净注射液同时孵育EPCs),应用四甲基偶氮唑盐(MTT )比色法测定EPCs 的增殖和黏附能力[吸光度(A)值].结果 与空白对照组比较,LPS 组EPCs 增殖、黏附能力均显著降低(增殖能力:0.1077±0.0026 比0.1248±0.0037,黏附能力:0.0647±0.0020 比0.0770±0.0052,均P＜0.05);与LPS 组比较,不同浓度血必净组EPCs 增殖能力均明显提高,且呈剂量-效应关系(均P＜0.05),EPCs 黏附能力有所改善,但未见明显剂量-效应关系,以血必净1 组25 g/L 浓度升高更显著(0.0855±0.0030,P＜0.05).结论 单纯LPS 刺激可导致EPCs 增殖、黏附能力下降;血必净注射液可使受损的EPCs 增殖、黏附能力改善,其中增殖能力随血必净注射液剂量增加而增强,而黏附能力与血必净注射液剂量间未见明显关系.提示血必净注射液可不同程度中和LPS 对EPCs 的损伤,在革兰阴性杆菌所致脓毒症早期使用血必净注射液可能有更好的效果.%Objective To investigate the effect of Xuehijing injection at different concentrations on the proliferation and adhesion of endothelial progenitor cells (EPCs) obtained from human umbilical venous blood incubated by lipopolysaccharide (LPS) in vitro. Methods After being resuscitated, cultured and passaged, EPCs were assessed as differentiating cells, inoculated into the cultural plate, and divided equally into four groups ; control group [cultivated with microvascular endothelial growth medium-2 (EGM-2MV) fluid ], LPS group ( LPS 100 μg/L) and Xuebijing 1st group (first incubated by LPS for 24 hours then added 5, 25, 50 g/L Xuebijing injection ) and Xuebijing 2nd group (incubated with LPS and 5, 25, 50 g/L Xuebijing injection for 43 hours). The influences of LPS and Xuebijing injection on EPCs' proliferation and adhesion ( absorbanee (A) value] were evaluated by four methyl thiazolyl tetrazolium salt (MTT) assay. Results Compared with the control group, the proliferation and adhesion of EPCs in LPS group were declined significantly (the proliferation : 0.1077±0.0026 vs. 0.1248±0.0037, the adhesion ; 0.0647±0.0020 vs. 0.0770±0.0052. both P＜0.05). Compared with the LPS group, Lite proliferation of EPCs were improved when Xuebijing injection was added, presenting dose-effect relationship (all P＜0.05) ; the adhesion of EPCs was also improved but no dose-effect relationship was seen, and the most significant improvement was in the group where 25 g/L Xuebijing injection was added in the Xuebijing 1st group (0.0855±0.0030, P＜ 0.05). Conclusion LPS can decrease the proliferation and adhesion of EPCs in certain degree. Xuebijing injection can improve the proliferation and adhesion of injured EPCs. whose proliferation is increased along with the increase of the dosage of Xuebijing injection, while their adhesion has no such dose-effect relationship, suggesting that Xuebijing injection have certain degree of action to neutralize the EPCs1 injury by LPS. Early application of this injection can make the treatment of sepsis induced by Gram-negative bacteria more effective.