PPARγ受体激动剂通过HIF-1α/BNIP3信号通路减轻脑出血模型大鼠继发性损伤的机制研究

Mechanism of PPARγReceptor Agonist in Reducing Secondary Injury of Cerebral Hemorrhage Model Rats Through HIF-1α/BNIP3 Signaling Pathway

目的探讨过氧化物酶体增殖物激活受体γ(PPARγ)受体激动剂通过低氧诱导因子1α(HIF-1α)/Bcl2/腺病毒E1B相互作用蛋白3(BNIP3)信号通路减轻脑出血模型大鼠继发性损伤的机制。方法本实验时间为2021年7月—2022年7月。选取健康SPF级雄性SD大鼠95只。选取10只大鼠进行模型筛选,选取25只大鼠用于确定指标检测时间。选取30只大鼠,随机分为A、B、C、D、E、F组,每组5只。A组大鼠在改良脑出血模型制备过程中注入0.9%氯化钠注射液代替自体血,建模前给予0.9%氯化钠溶液灌胃,建模后给予0.9%氯化钠溶液灌胃、腹腔注射。B、C、D、E、F组大鼠制备改良脑出血模型,其中B组大鼠建模前给予0.9%氯化钠溶液灌胃,建模后给予0.9%氯化钠溶液灌胃、腹腔注射;C组大鼠建模前给予0.9%氯化钠溶液灌胃,建模后给予罗格列酮(RSG)灌胃、0.9%氯化钠溶液腹腔注射;D组大鼠建模前给予RSG灌胃,建模后给予RSG灌胃、0.9%氯化钠溶液腹腔注射;E组大鼠建模前给予0.9%氯化钠溶液灌胃,建模后给予0.9%氯化钠溶液灌胃、2-甲氧基雌二醇(2ME2)腹腔注射;F组大鼠建模前给予RSG灌胃,建模后给予RSG灌胃、2ME2腹腔注射。比较六组大鼠建模后24、48、72 h改良神经功能缺损评分(mNSS),建模后72 h脑血肿周围组织HIF-1α、微管相关蛋白1轻链3(LC3)、BNIP3表达水平。选取30只大鼠,随机分为A、B、C、D、E、F组,每组5只。各组干预方法同前。比较六组大鼠建模后72 h脑血肿周围组织氧化损伤产物[8-羟基脱氧鸟苷(8-OHdG)、过氧化氢酶(CAT)、丙二醛(MDA)、蛋白质羰基(PCO)]含量。结果A组大鼠mNSS为0。建模后24 h,D组大鼠mNSS低于F组(P<0.05);建模后48 h,C、D组大鼠mNSS低于B组,F组大鼠mNSS高于D组(P<0.05);建模后72 h,D组大鼠mNSS低于B组,F组大鼠mNSS高于D组(P<0.05)。建模后72 h,B组大鼠脑血肿周围组织HIF-1α、LC3、BNIP3表达水平高于A、E组,低于C、D组(P<0.05);建模后72 h,D组大鼠脑血肿周围组织HIF-1α、LC3、BNIP3表达水平高于C、F组(P<0.05);建模后72 h,E组大鼠脑血肿周围组织HIF-1α、LC3、BNIP3表达水平低于F组(P<0.05)。建模后72 h,E组大鼠脑血肿周围组织8-OHdG含量高于B、F组,脑血肿周围组织CAT含量低于B组(P<0.05)。结论PPARγ受体激动剂能有效减轻脑出血模型大鼠继发性损伤,其机制可能与HIF-1α/BNIP3信号通路介导的自噬水平升高、DNA损伤产物生成减少有关。

Objective To investigate the mechanism of peroxisome proliferator-activated receptorγ(PPARγ)receptor agonist in reducing secondary injury of cerebral hemorrhage model rats through hypoxia inducible factor 1-alpha(HIF-1α)/Bcl2-adenovirus E1B 19-kDa interacting protein 3(BNIP3)signaling pathway.Methods The experimental period was from July 2021 to July 2022.A total of 95 healthy SPF male SD rats were selected.Ten rats were selected for model screening,and 25 rats were selected to determine the index detection time.A total of 30 rats were randomly divided into groups A,B,C,D,E and F,with 5 rats in each group.Rats in group A were injected with 0.9%sodium chloride injection instead of autologous blood during the preparation of modified cerebral hemorrhage model,and were given 0.9%sodium chloride solution by gavage before modeling,0.9%sodium chloride solution by gavage and intraperitoneal injection after modeling.The rats in groups B,C,D,E and F were used to prepare the modified cerebral hemorrhage model.The rats in group B were given 0.9%sodium chloride solution by gavage before modeling,and 0.9%sodium chloride solution by gavage and intraperitoneal injection after modeling.Rats in group C were given 0.9%sodium chloride solution by gavage before modeling,and rosiglitazone(RSG)by gavage and 0.9%sodium chloride solution by intraperitoneal injection after modeling.Rats in group D were given RSG by gavage before modeling,and RSG by gavage and 0.9%sodium chloride solution by intraperitoneal injection after modeling.Rats in group E were given 0.9%sodium chloride solution by gavage before modeling,and 0.9%sodium chloride solution by gavage and 2-methoxyestradiol(2ME2)by intraperitoneal injection after modeling.The rats in group F were given RSG by gavage before modeling,and RSG by gavage and 2ME2 by intraperitoneal injection after modeling.The modified Neurological Severity Score(mNSS)at 24,48 and 72 h after modeling,the expression levels of HIF-1α,microtubule-associated protein 1 light chain 3(LC3),BNIP3 in the tissues around the hematoma at 72 h after modeling were compared among the six groups.A toal of 30 rats were randomly divided into groups A,B,C,D,E and F,with 5 rats in each group.The intervention methods of each group were the same as before.The contents of oxidative damage products[8-hydroxy-2'-deoxyguanosine(8-OHdG),catalase(CAT),malondialdehyde(MDA)and protein carbonyl(PCO)]in the tissues around the hematoma at 72 h after modeling were compared among the six groups.Results The mNSS of rats in group A was 0.At 24 h after modeling,the mNSS of rats in group D was lower than that in group F(P<0.05).At 48 h after modeling,the mNSS of rats in groups C and D was lower than that in group B,and the mNSS of rats in group F was higher than that in group D(P<0.05).At 72 h after modeling,the mNSS of rats in group D was lower than that in group B,and the mNSS of rats in group F was higher than that in group D(P<0.05).At 72 h after modeling,the expression levels of HIF-1α,LC3 and BNIP3 in the tissues around the hematoma of rats in group B were higher than those in groups A and E,and those were lower than those in groups C and D(P<0.05).At 72 h after modeling,the expression levels of HIF-1α,LC3 and BNIP3 in the tissues around the hematoma of rats in group D were higher than those in groups C and F(P<0.05).At 72 h after modeling,the expression levels of HIF-1α,LC3 and BNIP3 in the tissue around the hematoma of rats in group E were lower than those in group F(P<0.05).At 72 h after modeling,the content of 8-OHdG in the tissues around the hematoma of rats in group E was higher than that in groups B and F,and the content of CAT was lower than that in group B(P<0.05).Conclusion PPARγreceptor agonist can effectively reduce the secondary injury in rats with cerebral hemorrhage model.The mechanism may be related to the increase of autophagy level and the decrease of DNA damage products mediated by HIF-1α/BNIP3 signaling pathway.