基于 PERK/ATF4/CHOP 通路探讨牛蒡子苷元对心肌梗死大鼠 Treg/Th17 平衡及心功能的影响

Exploring the effects of arctigenin on Treg/Th17 balance and cardiac function in rats with myocardial infarction based on the PERK-ATF4-CHOP pathway

关键词:  牛蒡子苷元  PERK-ATF4-CHOP通路  心肌梗死  Treg/Th17平衡  心功能

Key words:  arctigenin  PERK-ATF4-CHOP pathway  myocardial infarction  Treg/Th17 balance  cardiac function

摘要:

[目的] 探讨牛蒡子苷元（ARC）是否可调控PKR样内质网激酶（PERK）/激活转录因子4（ATF4）/C/EBP同源蛋白（CHOP）通路影响心肌梗死（MI）大鼠调节性T细胞（Treg）/辅助性T细胞17（Th17）平衡和心功能。[方法] 将大鼠随机分为对照（Con）组、MI组、ARC低剂量（ARC-L）组、ARC高剂量（ARC-H）组、ARC-H+CCT020312（PERK激活剂）组。检测大鼠心功能、血清乳酸脱氢酶（LDH）、白细胞介素（IL-10、IL-17A、IL-22）水平和心肌损伤标志物肌酸激酶同工酶MB（CK-Mb）、心肌肌钙蛋白I（cTnI）、心肌肌钙蛋白T（cTnT）水平、外周血中Th17和Treg细胞比例、心肌梗死面积百分比；观察大鼠心脏组织形态；检测心肌组织细胞凋亡、裂解的半胱氨酸蛋白酶3（Cleaved Caspase-3）和PERK/ATF4/CHOP通路蛋白表达水平。[结果] 与Con组比较，MI组大鼠心肌组织有明显损伤，左室收缩末期压（LVESP）、左室射血分数（LVEF）和左室短轴缩短率（LVFS）水平、IL-10水平、Treg细胞比例和Treg/Th17值降低，左室舒张末期压（LVEDP）水平、LDH、IL-17A和IL-22水平、CK-Mb、cTnI和cTnT水平、Th17细胞比例、MI面积百分比、细胞凋亡率、p-PERK/PERK、p-eIF2α/eIF2α、ATF4、CHOP和Cleaved Caspase-3蛋白表达水平升高（P<0.05）。与MI组比较，ARC-L组和ARC-H组大鼠心肌组织形态有明显改善，LVESP、LVEF和LVFS水平、IL-10水平、Treg细胞比例和Treg/Th17值升高，LVEDP水平、LDH、IL-17A和IL-22水平、CK-Mb、cTnI和cTnT水平、Th17细胞比例、心肌梗死面积百分比、细胞凋亡率、p-PERK/PERK、p-eIF2α/eIF2α、ATF4、CHOP和Cleaved Caspase-3蛋白表达水平降低（P<0.05）。CCT020312可降低ARC对MI大鼠的改善作用（P<0.05）。[结论] ARC可能通过抑制PERK-ATF4-CHOP通路来降低MI大鼠炎症、心肌细胞凋亡和心肌损伤，改善Treg/Th17平衡和心功能。

Abstract:

[Objective] To explore whether arctigenin（ARC） can regulate the PKR-like endoplasmic reticulum kinase（PERK）-activated transcription factor 4（ATF4）-C/EBP homologous protein（CHOP） pathway to affect the Regulatory T cells（Treg）/Helper T cell 17（Th17） balance and cardiac function in rats with myocardial infarction（MI）. [Methods] Rats were randomly separated into the control（Con） group，the MI group，the ARC low-dose（ARC-L） group，the ARC high-dose（ARC-H） group，and the ARC-H+CCT020312（PERK activator） group. The cardiac function of rats，the levels of serum Lactate dehydrogenase（LDH），interleukins（IL-10，IL-17A，IL-22），the levels of myocardial injury markers Creatine kinase isoenzyme MB（CK-Mb），cardiac troponin I（cTnI），and cardiac troponin T（cTnT），the proportions of Th17 and Treg cells in peripheral blood，and the percentage of myocardial infarction area were measured. The morphology of rat cardiac tissue was observed. The apoptosis of myocardial tissue cells and the expression levels of Cleaved Caspase-3 and PERK/ATF4/CHOP pathway proteins were measured. [Results] Compared with the Con group，the myocardial tissue of rats in the MI group was clearly damaged，the levels of LVESP，LVEF and LVFS，the level of IL-10，the proportion of Treg cells，and the value of Treg/Th17 were lower，while the level of LVEDP，the levels of LDH，IL-17A and IL-22，the levels of CK-Mb，cTnI and cTnT，proportion of Th17 cells，percentage of myocardial infarction area，apoptosis rate，the expression levels of p-PERK/PERK，p-eIF2α/eIF2α，ATF4，CHOP and Cleaved Caspase-3 proteins were higher（P<0.05）. Compared with the MI group，the myocardial tissue morphology of rats in the ARC-L group and the ARC-H group was clearly improved，the levels of LVESP，LVEF and LVFS，the level of IL-10，the proportion of Treg cells，and the value of Treg/Th17 were higher，while the level of LVEDP，the levels of LDH，IL-17A and IL-22，the levels of CK-Mb，cTnI and cTnT，proportion of Th17 cells，percentage of myocardial infarction area，apoptosis rate，the expression levels of p-PERK/PERK，p-eIF2α/eIF2α，ATF4，CHOP and Cleaved Caspase-3 proteins were lower（P<0.05）. CCT020312 was able to reduce the improvement effect of ARC on MI rats（P<0.05）. [Conclusion] ARC may reduce inflammation，cardiomyocyte apoptosis and myocardial injury in MI rats，and ameliorate Treg/Th17 balance and cardiac function by inhibiting the PERK-ATF4-CHOP pathway.