摘要
目的研究由桃红四物汤及二陈汤化裁而成的祛痰清瘀方对氧化低密度脂蛋白(ox-LDL)诱导人原代主动脉内皮细胞(HAoEC)病理性活化及增殖的抑制能力和机制。方法 60只大鼠,高脂饲养结合链尿菌素(STZ)建立大鼠糖尿病大血管病变模型,将大鼠随机分为对照组、模型组、祛痰清瘀方组(2.5 g/kg)、辛伐他汀组(20 mg/kg),各10只;比较四组大鼠血糖和血脂水平及主动脉脂质含量。以20 mg/L的ox-LDL刺激HAoEC建立活化增殖模型,分为对照组、ox-LDL组和ox-LDL+祛痰清瘀方组,各10只。比较四组大鼠内皮Yes相关蛋白(YAP)表达水平及丝裂原活化蛋白激酶1/2(Erk1/2)磷酸化水平;比较五组HAoEC增殖速率、细胞中LATS1/2和YAP磷酸化水平,细胞中YAP靶基因CTGF、CYR61和ANKRD1表达水平。结果模型组大鼠空腹血糖(FG)为(21.31±3.93)mmol/L、甘油三酯(TG)为(3.62±1.37)mmol/L、总胆固醇(TC)为(7.51±2.17)mmol/L和低密度脂蛋白胆固醇(LDL-C)为(5.07±1.46)mmol/L均显著高于对照组的(5.42±0.62)、(0.84±0.06)、(1.78±0.25)、(0.38±0.15)mmol/L,高密度脂蛋白胆固醇(HDL-C)的(0.36±0.14)mmol/L低于对照组的(0.81±0.22)mmol/L,差异有统计学意义(P<0.05);模型组大鼠血清TG水平显著高于祛痰清瘀方组,差异有统计学意义(P<0.05);模型组大鼠血清TG、TC和LDL-C水平均显著高于辛伐他汀组,差异有统计学意义(P<0.05)。模型组大鼠主动脉TG和TC含量均显著高于对照组、祛痰清瘀方组和辛伐他汀组,差异有统计学意义(P<0.01或<0.05)。模型组大鼠主动脉中YAP的表达水平及ERK1/2的磷酸化水平均显著高于对照组、祛痰清瘀方组和辛伐他汀组,差异有统计学意义(P<0.01)。ox-LDL组增殖速率均高于对照组、祛痰清瘀方组和辛伐他汀组,低于祛痰清瘀方+ERK过表达组,差异有统计学意义(P<0.01)。ox-LDL组细胞中LATS1/2和YAP的磷酸化水平显著低于对照组、祛痰清瘀方组和辛伐他汀组, Erk1/2的磷酸化比例显著高于对照组、祛痰清瘀方组和辛伐他汀组,差异均有统计学意义(P<0.01);祛痰清瘀方+ERK过表达组LATS1/2和YAP的磷酸化水平低于祛痰清瘀方组,差异均有统计学意义(P<0.01);各组MST1/2磷酸化水平比较,差异无统计学意义(P>0.05)。ox-LDL组细胞中YAP靶基因CTGF、CYR61和ANKRD1表达水平显著高于对照组、祛痰清瘀方组和辛伐他汀组,差异均有统计学意义(P<0.01);祛痰清瘀方+ERK过表达组YAP靶基因CTGF、CYR61和ANKRD1的表达水平显著高于祛痰清瘀方组,差异均有统计学意义(P<0.01)。结论祛痰清瘀方通过干扰丝裂原信号转导蛋白Erk1/2,阻碍YAP活化,抑制内皮细胞病理性增殖介导的糖尿病大血管病变。
Objective To study the inhibitory ability of Qutan Qingyu decoction, which is made from Taohong Siwu decoction and Erchen decoction, on the pathological activation and proliferation of human primary aortic endothelial cells(HAoEC) induced by oxidized low-density lipoprotein(ox-LDL) And mechanism. Methods 60 rats, high-fat feeding combined with streptozotocin(STZ) to establish a rat model of diabetic macrovascular disease, the rats were randomly divided into control group, model group, Qutan Qingyu decoction group(2.5 g/kg), simvastatin group(20 mg/kg), with 10 rats in each group;The blood glucose, blood lipid levels and the aortic lipid content of the four groups were compared. HAoEC was stimulated with 20 mg/L ox-LDL to establish an activated proliferation model, which was divided into control group, ox-LDL group and ox-LDL+Qutan Qingyu decoction group, with 10 rats each group. The expression of Yes-associated protein(YAP) and the phosphorylation level of mitogen-activated protein kinase 1/2(Erk1/2) in the four groups were compared;the proliferation rate, LATS1/2 and Yap phosphorylation levels, and the expression levels of Yap target genes CTGF, Cyr61 and ANKRD1 were compared among the five groups. Results The fasting glucose(FG)(21.31±3.93) mmol/L, triglyceride(TG)(3.62±1.37) mmol/L, total cholesterol(TC)(7.51±2.17) mmol/L and low-density lipoprotein cholesterol(LDL-C)(5.07±1.46) mmol/L of model group were significantly higher than(5.42±0.62) mmol/L,(0.84±0.06) mmol/L,(1.78±0.25) mmol/L,(0.38±0.15) mmol/L of the control group, and high-density lipoprotein cholesterol(HDL-C)(0.36±0.14) mmol/L was lower than(0.81±0.22) mmol/L of the control group, and the difference was statistically significant(P<0.05);the serum TG of model group was significantly higher than that of Qutan Qingyu decoction group, and the difference was statistically significant(P<0.05);the serum TG, TC and LDL-C of model group were significantly higher than those of simvastatin group, and the difference was statistically significant(P<0.05). The TG and TC content in aorta of model group were significantly higher than those of control group, Qutan Qingyu decoction group and simvastatin group, and the difference was statistically significant(P<0.01 or <0.05). The YAP expression and phosphorylation level of ERK 1/2 in aorta of model group were significantly higher than those of the control group, Qutan Qingyu decoction group and simvastatin group, and the difference was statistically significant(P<0.01). The proliferation rate of ox-LDL group was higher than that of control group, Qutan Qingyu decoction group and simvastatin group, and lower than that of Qutan Qingyu decoction + ERK over-expression group, and the difference was statistically significant(P<0.01). The phosphorylation levels of LATS1/2 and YAP in the cells of the ox-LDL group were significantly lower than those of the control group, Qutan Qingyu decoction group and the simvastatin group, and phosphorylation ratio of Erk1/2 was significantly higher than that of the control group, Qutan Qingyu decoction group, and the difference was statistically significant(P<0.01);the phosphorylation levels of LATS1/2 and YAP in the Qutan Qingyu decoction + ERK over-expression group were lower than those in the Qutan Qingyu decoction group, and the differences were statistically significant(P<0.01);there was no statistically significant difference in the phosphorylation level of MST1/2 in each group(P>0.05). The expression levels of YAP target genes CTGF, CYR61 and ANKRD1 in the cells of the ox-LDL group were significantly higher than those of the control group, Qutan Qingyu decoction group and simvastatin group, and the differences were statistically significant(P<0.01);the expression levels of YAP target genes CTGF, CYR61 and ANKRD1 in the Qutan Qingyu decoction+ERK over-expression group were significantly higher than those in the Qutan Qingyu decoction group, and the differences were statistically significant(P<0.01). Conclusion Qutan Qingyu decoction inhibits the activation of YAP by interfering with mitogen signal transduction proteins Erk1/2, and thus inhibits diabetic macroangiopathy.
作者
寇鑫晖
赵恒侠
陈军
李惠林
KOU Xin-hui;ZHAO Heng-xia;CHEN Jun(Shenzhen Traditional Chinese Medicine Hospital,Shenzhen 518033,China)
出处
《中国实用医药》
2020年第34期202-206,共5页
China Practical Medicine
基金
深圳市科技创新委员会面上项目(项目编号:JCYJ20180302173605034)
深圳市卫生健康委员会博士创新项目(项目编号:SZBC2018004)
深圳市医疗卫生三名工程(项目编号:SZSM201512043)。
关键词
桃红四物汤
二陈汤
祛痰清瘀方
糖尿病大血管并发症
内皮细胞
Taohong Siwu decoction
Erchen decoction
Qutan Qingyu decoction
Diabetic macroangiopathy
Aortic endothelial cells
作者简介
通讯作者:李惠林。
