辅酶Q10对脓毒血症休克大鼠血管平滑肌细胞功能的影响

Effect of coenzyme Q10 on function of vascular smooth muscle cell in rats with septic shock

目的探讨辅酶Q10对脓毒血症休克大鼠血管平滑肌细胞功能的影响及其作用机制研究。方法采用SD大鼠尾静脉注射脂多糖(LPS)的方法建立脓毒症休克动物模型,分离取出主动脉,0.2%胶原酶消化分离平滑肌细胞(SMCs),自然纯化及差速贴壁纯化血管平滑肌细胞,将分离后的SMC分别接种到96孔板中,分为4组,分别为正常对照组、正常对照组+辅酶Q10、LPS组、LPS+辅酶Q10组,分别检测4组的增殖和氧化应激产物产生情况,在6、12、24、48 h后分别检测正常对照组、正常对照组+辅酶Q10、LPS组、LPS+辅酶Q10组SMCs丙二醛水平,在48 h后检测α-肌动蛋白(α-actin)、骨桥蛋白(OPN)的相对表达量。结果随时间推移,LPS组中SMC细胞凋亡和氧化应激产物均较正常对照组增多,在48 h时达到峰值,分别增多11.4倍和5.6倍;LPS+辅酶Q10组中氧化应激产物较LPS组减少近50%;在48 h时辅酶Q10在正常培养环境下对SMC的α-actin、OPN蛋白的相对表达量差异无统计学意义(P>0.05),而对脓毒症休克SD大鼠分离的SMC可以明显减低氧化应激产物的生成及上调α-actin蛋白的相对表达量近1.57倍,下调OPN蛋白的相对表达量1.17倍。结论由LPS所致的脓毒症休克的发病机制与血管中膜平滑肌细胞表型改变密切相关,辅酶Q10可明显减低氧化应激产物的生成及上调α-actin蛋白的相对表达量,缓解了因LPS所致脓毒血症休克的血管病理化转变。收缩型平滑肌细胞是正常情况下血管壁的重要组成部分,而疾病病理血管则表现为合成型,为脓毒血症休克的病理性血管形成提供理论基础。

Objective To investigate the effect of coenzyme Q10 on the function of vascular smooth muscle cell(VSMC)in septic rats and its mechanisms.Methods An animal model of sepsis shock was established by tail vein injection of endotoxin lipopolysaccharide(LPS).The aorta from SD rat was separated and smooth muscle cells were isolated with collagenase.The cells were collected by the method of natural purification and purification of sticking wall at different speed.The proliferation and oxidative stress production was respectively detected by enzyme standard instrument in four groups,including control group,control group+coenzyme Q10,LPS group,LPS group+coenzyme Q10.The concentration of malondialdehyde(MDA)was detected at 6,12,24,48 h.The protein expression of α-SM-actin and OPN was detected after 48 h.Results Compared with the control group,the number of cell apoptosis and oxidative stress production in the LPS group,peaking at 48 h,were increased by 11.4 times and 5.6 times,respectively;on the contrary,compared with LPS group,the oxidative stress production were reduced nearly 50% in LPS group+coenzyme Q10;In control group,The protein expression ofα-SM-actin and OPN had no statistical differences in VSMC treated with coenzyme Q10 at 48 h(P>0.05).In LPS group,the protein expression of α-SM-actin obviously increased 1.57 times in VSMC treated with coenzyme Q10 at 48 h,except for the protein expression of OPN decreasing 1.17 times.Conclusions The pathogenesis of sepsis shock caused by lipopolysaccharide(LPS)is closely related to phenotypic changes of vascular smooth muscle cell;Coenzyme Q10 can significantly reduce the production of oxidative stress products and increase the relative expression of α-actin protein,and alleviate the vascular pathological transformation of sepsis shock caused by LPS.Constrictive smooth muscle cells are an important part of the vascular wall under normal conditions,while disease pathological blood vessels are shown as synthetic,which provides a theoretical basis for pathological vascular formation in sepsis shock.