摘要
探讨丹参酮Ⅱ_(A)(tanshinoneⅡ_(A),TAⅡ_(A))对抗磷脂抗体综合征(antiphospholipid syndrome,APS)患者血清干预人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)的活化因子表达的调控作用以及对磷脂酰肌醇3激酶(phosphoinositide3-kinase,PI3K)/蛋白激酶B(protein kinase B,PKB,又名Akt)信号通路的影响。通过体外培养HUVEC,分为单纯培养基组、空白对照组、APS模型组、APS+LY5组、APS+LY10组、APS+LY20组、APS+TAⅡ_(A)5组、APS+TAⅡ_(A)10组、APS+TAⅡ_(A)20组、APS+TAⅡ_(A)10+LY10组,检测不同浓度LY294002和TAⅡ_(A)对HUVEC分泌白细胞介素-6(interleukin 6,IL-6)、白细胞介素-8(interleukin 8,IL-8)、单核细胞趋化蛋白-1(monocyte chemoattractant protein,MCP-1)的影响,qPCR测定其对膜联蛋白A2(annexin A2,ANXA2)、PI3K、Akt、E-钙黏素(E-cadherin,E-cad)mRNA表达的影响,Western blot法检测其对ANXA2、p-PI3K/PI3K、p-Akt/Akt、E-cad蛋白表达的影响。发现与APS模型组比较,APS+TAⅡ_(A)10组分泌IL-6、MCP-1量降低,分泌IL-8量增多,差异具有统计学意义,而且随着TAⅡ_(A)剂量的增加,呈浓度依赖性;APS+TAⅡ_(A)10组ANXA2、PI3K、Akt、E-cad mRNA及蛋白表达水平增加,差异具有统计学意义(P<0.05或P<0.01),而且随着TAⅡ_(A)剂量的增加,呈浓度依赖性。表明APS患者血清可以导致HUVEC的ANXA2、PI3K、Akt、E-cad mRNA和蛋白表达水平下降;IL-6、MCP-1分泌量增多,IL-8分泌量降低;APS患者血清可活化血管内皮细胞。而一旦PI3K/Akt信号通路被阻断之后,ANXA2、E-cad的mRNA及蛋白的表达均出现明显下降,IL-6、MCP-1分泌量明显增多,IL-8分泌量明显降低。这表明TAⅡ_(A)通过激活PI3K/Akt信号通路调控APS患者血管内皮细胞的活化。
The present study investigated the regulatory effect of tanshinoneⅡ_(A)(TAⅡ_(A)) on activator expression in human umbilical vein endothelial cells(HUVECs)and the effect on the phosphoinositide 3-kinase(PI3 K)/protein kinase B(Akt)signaling pathway in patients with antiphospholipid syndrome(APS).HUVECs cultured in vitro were divided into a medium group,a blank control group,an APS model group,an APS+LY5 group,an APS+LY10 group,an APS+LY20 group,an APS+TAⅡ_(A)5 group,an APS+TAⅡ_(A)10 group,an APS+TAⅡ_(A)20 group,and an APS+TAⅡ_(A)10+LY10 group.The effects of LY294002 and TAⅡ_(A)at different concentrations on the secretion of interleukin-6(IL-6),interleukin-8(IL-8),and monocyte chemoattractant protein-1(MCP-1)by HUVECs were investigated.The effects on the mRNA expression of annexin A2(ANXA2),PI3 K,Akt,and E-cadherin(E-cad)were detected by quantitative polymerase chain reaction(qPCR),and Western blot was used to determine the effects on the protein expression of ANXA2,p-PI3 K/PI3 K,p-Akt/Akt,and E-cad.The results revealed that compared with the APS model group,the APS+TAⅡ_(A)10 group showed statistically reduced IL-6 and MCP-1 and increased IL-8 in a concentration-dependent manner with the increase in TAⅡ_(A) dose,while the APS+TAⅡ_(A)10 group showed increased mRNA and protein expression of ANXA2,PI3 K,Akt,and E-cad(P<0.05 or P<0.01)in a concentration-dependent manner with the increase in TAⅡ_(A) dose.The findings indicated that the serum of APS patients could lead to the decreased mRNA and protein expression levels of ANXA2,PI3 K,Akt,and E-cad in HUVECs,increased secretion of IL-6 and MCP-1,and reduced secretion of IL-8,and activate vascular endothelial cells.In contrast,once the PI3 K/Akt signaling pathway was blocked,the mRNA and protein expression of ANXA2 and E-cad significantly decreased,IL-6 and MCP-1 secretion significantly increased,and IL-8 secretion was significantly reduced.It suggests that TAⅡ_(A)regulates the activation of vascular endothelial cells in APS patients by activating
作者
童星丽
谈勇
殷燕云
郭银华
TONG Xing-li;TAN Yong;YIN Yan-yun;GUO Yin-hua(Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210004,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2022年第13期3589-3596,共8页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(81873333)
江苏省科教强卫工程“中医妇科学临床医学中心”项目(YXZXB2016006)。
