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白藜芦醇抑制血管紧张素Ⅱ/TLR4/NF-KB调控鼠血管平滑肌细胞炎症和氧化应激损伤作用

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摘要:

目的 探讨白藜芦醇(RES)抑制血管紧张素Ⅱ(AngⅡ)/TLR4/NF-KB调控大鼠血管平滑肌细
胞炎症和氧化应激损伤的作用。方法 采用大鼠胸主动脉平滑肌A7r5细胞株为主要受试物,分为对照组、RES
组(30 μmol/L)、AngⅡ组(10-4 μmol/L)和实验组(30 μmol/L RES预处理2 h+10-4 μmol/L AngⅡ)。CCK-8
法检测A7r5细胞的存活率;ELISA法检测细胞上清液中TNF-α和IL-6表达;采用试剂盒检测ROS和iNOS活
性;Western blot法检测A7r5细胞AngⅡ、TLR4、P-P65,P-IKB蛋白表达水平。结果 与对照组相比,AngⅡ组
A7r5细胞株存活率显著升高,RES组和实验组A7r5细胞株存活率显著降低(P<0.05);AngⅡ组A7r5细胞株
TNF-α和IL-6表达显著升高,ROS和iNOS活性显著升高,AngⅡ、TLR4、P-P65,P-IKB蛋白含量显著升高,
RES组和实验组TNF-α和IL-6表达显著降低,ROS和iNOS活性显著降低,AngⅡ、TLR4、P-P65,P-IKB蛋白
含量显著升高(P<0.05)。与AngⅡ组相比,RES组和实验组A7r5细胞株存活率显著降低,实验组A7r5细胞
株存活率下降更为明显(P<0.05);AngⅡ组A7r5细胞株TNF-α和IL-6表达显著降低,ROS和iNOS活性显著
降低,AngⅡ、TLR4、P-P65,P-IKB蛋白含量显著降低,RES组和实验组TNF-α和IL-6表达显著降低,ROS
和iNOS活性显著降低,AngⅡ、TLR4、P-P65,P-IKB蛋白含量显著降低(P<0.05)。结论 白藜芦醇可有效
缓解大鼠血管平滑肌细胞的炎症和氧化应激损伤,主要通过抑制血管紧张素Ⅱ/TLR4/NF-KB信号通路。

Abstract:

Objective To investigate the effects of resveratrol (RES) on angiotensin Ⅱ/TLR4/NF-KB in regulating
vascular smooth muscle cell inflammation and oxidative stress injury in rats. Methods Rat thoracic aortic smooth muscle
A7r5 cell line was used as the main test substance, which was divided into control group, RES group (30 μmol/L), Ang
Ⅱ group (10-4 μmol/ L) and experimental group (30 μmol/L RES pretreatment for 2 hours+10-4 μmol/L Ang Ⅱ). The
survival rate of A7r5 cells was detected by CCK-8 method. The expression of TNF-α and IL-6 in cell supernatant was
detected by ELISA. The activities of ROS and iNOS were detected by kit. Western blot method was used to detect AngⅡ,
TLR4, P-P65 of A7r5 cells , P-IKB protein expression level. Result Compared with the control group, the survival rate of
the A7r5 cell line in the AngⅡ group was significantly increased, and the survival rate of the A7r5 cell line in the RES group
and the experimental group was significantly reduced (P<0.05); the expression of TNF-α and IL-6 in the A7r5 cell line in
the AngⅡ group was significantly increased. High, ROS and iNOS activities were significantly increased, AngⅡ, TLR4,
P-P65, P-IKB protein content was significantly increased, TNF-α and IL-6 expressions were significantly reduced in
RES and experimental groups, and ROS and iNOS activities were significantly reduced. AngⅡ, TLR4, P-P65, and P-IKB
protein levels were significantly increased (P<0.05). Compared with the AngⅡ group, the survival rate of the A7r5 cell line
in the RES group and the experimental group was significantly reduced, and the survival rate of the A7r5 cell line in the
experimental group was significantly reduced (P<0.05); the expression of TNF-α and IL-6 in the A7r5 cell line of the Ang
Ⅱ group was significantly lower Decreased, ROS and iNOS activities were significantly reduced, AngⅡ, TLR4, P-P65, and
P-IKB protein contents were significantly reduced, TNF-α and IL-6 expressions were significantly reduced in RES and
experimental groups, ROS and iNOS activities were significantly reduced, and AngⅡ, TLR4, P-P65, and P-IKB protein
levels were significantly reduced (P<0.05). Conclusion Resveratrol can effectively alleviate inflammation and oxidative
stress injury of vascular smooth muscle cells in rats, mainly by inhibiting the angiotensin Ⅱ/TLR4/NF-KB signaling pathway.

基金项目:

辽宁省科技厅科学技术计划项目(2011225017)

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