激活PPAR-γ对脑缺血-再灌注损伤中炎症反应的影响  

Effects of PPAR-γ activation on inflammation in cerebral ischemia and reperfusion injury

作　　者：黄炎 彭拓 陈诚[1] 夏萍萍[1] 张帆[1] 李龙艳[1] 叶治[1] 贺正华[1] HUANG Yan;PENG Tuo;CHEN Cheng;XIA Pingping;ZHANG Fan;LI Longyan;YE Zhi;HE Zhenghua(Department of Anesthesiology,Xiangya Hospital Affiliated to Central South University,Changsha 410000,China)

机构地区：[1]中南大学湘雅医院麻醉科,长沙市410000 [2]武汉大学中南医院麻醉科

出　　处：《临床麻醉学杂志》2020年第7期685-690,共6页Journal of Clinical Anesthesiology

基　　金：国家自然科学基金(81771422);湖南省自然科学基金(2019JJ50969)。

摘　　要：目的探究激活过氧化物酶体增殖物激活受体-γ(PPAR-γ)对脑缺血-再灌注损伤(CIRI)与脂多糖(LPS)诱导炎症反应的影响。方法在动物实验中,SD雄性大鼠40只,体重250~270 g,随机分成四组:假手术组(S组)、缺血-再灌注组(IR组)、二甲基亚砜(DMSO)+缺血-再灌注组(DIR组)和吡格列酮+缺血-再灌注组(PIR组)。其中S组进行假手术处理,IR组进行脑中动脉缺血(MCAO)建模处理,DIR组给予DMSO,随后进行MCAO造模处理。PIR组给予吡格列酮,随后进行MCAO造模处理。在细胞实验中,通过慢病毒转染BV2细胞使PPAR-γ过表达,将细胞培养孔随机分为三组:空白对照组(B组)、阴性对照组(L1组)、过表达组(L2组)。其中B组未做处理,L1组转染PPAR-γ阴性病毒,L2组转染PPAR-γ阳性病毒。之后将细胞培养孔随机分为四组:正常对照组(C组)、正常细胞+LPS处理组(L组)、阴性对照+LPS处理组(LL1组)和过表达+LPS处理组(LL2组)。其中C组未做处理,L组用LPS处理,LL1组转染阴性病毒并确定转染率后使用LPS处理,LL2组转染阳性病毒并确定转染率后用LPS处理。各组大鼠使用Zea Longa评分标准进行神经行为学评分,TTC染色比较脑梗死容积百分比大小,采用ELISA法检测四组大鼠和四组BV2细胞IL-1β和IL-18浓度,Western blot法检测PPAR-γ、活化的半胱天冬酶1(caspase-1)和NOD样受体家族3(NLRP3)蛋白相对含量。结果与IR组比较,PIR组大鼠行为学评分明显降低,脑梗死容积百分比明显减小,PPAR-γ的蛋白相对含量明显增高,活化的caspase-1、NLRP3蛋白相对含量明显降低,IL-1β和IL-18浓度明显降低(P<0.05)。与B组比较,L2组的PPAR-γ蛋白相对含量明显增高。与C组比较,LL2组活化的caspase-1和NLRP3蛋白相对含量明显升高(P<0.05),IL-1β和IL-18浓度明显升高(P<0.05)。结论吡格列酮通过激活PPAR-γ减轻脑缺血-再灌注损伤,其机制是通过激活PPAR-γ,抑制NLRP3的表达,减轻炎症反应,进而使可能存在�Objective To investigate the effect of PPAR-γ activation on cerebral ischemia-reperfusion injury(CIRI) and lipopolysaccharide(LPS) induced cell inflammation. Methods Forty male SD rats, weighing 250-270 g, were randomly divided into 4 groups: sham group(group S), ischemia-reperfusion group(group IR), DMSO+ischemia-reperfusion group(group DIR) and pioglitazone+ischemia-reperfusion group(group PIR). BV2 cells transfected with lentivirus that overexpressed PPAR-γ were randomly divided into 3 groups: blank group(group B), negative-control group(group L1), and overexpression group(group L2). BV2 cells were then randomly divided into 4 groups: control group(group C), BV2+LPS group(group L), LV^- +LPS group(group LL1) and LV^+ +LPS group(group LL2). Neurological deficit scores were recorded in each group by using Zea Longa standard, and cerebral infarct volume percentage was measured by TTC staining. The expression of PPAR-γ, active caspase-1 and NLRP3 were detected by western blot, and the content of IL-1β and IL-18 were detected by ELISA in rats and BV2 cells. Results Compared with the group IR, the behavioral score of the group PIR was significantly reduced,the level of PPAR-γ in group PIR was significantly increased. Moreover, pioglitazone notably decreased infarct volume percentage and inhibited the level of NLRP3 and caspase-3 activation, as well as the content of IL-1β and IL-18(P<0.05). Compared with group B, the relative level of PPAR-γ protein in group L2 was significantly increased. Compared with group C, the levels of active caspase-1, NLRP3, IL-1β and IL-18 in group LL2 were significantly increased(P<0.05). Conclusion Pioglitazone relieved cerebral ischemia-reperfusion injury by the mechanisms involving in inhibiting NLRP3 activation,reduce inflammation,and subsequently reducing thepossible caspase-1-dependent pyroptosis through activating PPAR-γ.

关 键 词：过氧化物酶体增殖物激活受体-Γ 脑缺血-再灌注损伤 细胞焦亡 半胱天冬酶-1 

分 类 号：R743[医药卫生—神经病学与精神病学]