[1]胡跃强,陈 炜,祝美珍,等.清热化瘀方通过调控自噬相关基因P62/LC3保护大鼠脑缺血再灌注损伤*[J].陕西中医,2020,(4):429-433.
 HU Yueqiang,CHEN Wei,ZHU Meizhen,et al.Protective effect of Qingre Huayu prescription against cerebral ischemia-reperfusion injury by regulating autophagy-related gene P62/LC3 in rats[J].,2020,(4):429-433.
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清热化瘀方通过调控自噬相关基因P62/LC3保护大鼠脑缺血再灌注损伤*
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《陕西中医》[ISSN:1000-7369/CN:61-1281/TN]

卷:
期数:
2020年4期
页码:
429-433
栏目:
基础研究
出版日期:
2020-04-05

文章信息/Info

Title:
Protective effect of Qingre Huayu prescription against cerebral ischemia-reperfusion injury by regulating autophagy-related gene P62/LC3 in rats
文章编号:
DOI:10.3969/j.issn.10007369.2020.04.007
作者:
胡跃强1陈 炜1祝美珍2梁 妮1吴 林2唐 农2△
1.广西中医药大学第一附属医院(南宁 530023); 2.广西中医药大学(南宁 530001)
Author(s):
HU YueqiangCHEN WeiZHU Meizhenet al.
The First Affiliated Hospital of Guangxi University of TCM(Nanning 530023)
关键词:
脑缺血再灌注损伤 清热化瘀方 神经元 自噬 P62/LC3 大鼠
Keywords:
Qingre Huayu prescription Cerebral ischemia reperfusion Euronal Autophagy P62/LC3 Rats
分类号:
R743.31
文献标志码:
A
摘要:
目的:从自噬相关蛋白P62/LC3角度探讨清热化瘀方对大鼠脑缺血再灌注损伤的保护作用。方法:160只大鼠随机分为4组:假手术组(SO组)、模型组(I/R组)、清热化瘀颗粒组(QRHY组)和清开灵颗粒对照组(QKL组)。分别于再灌注后12 h、1 d、2 d、3 d取材(n=10)。线栓法制备大鼠局灶性脑缺血再灌注模型,分别采用透射电子显微镜观察神经元自噬及凋亡形态结构变化,qRT-PCR和Western blot观察P62及LC3 mRNA及其蛋白表达变化。结果:①大鼠脑缺血再灌注后激活自噬,再灌注后12 h至3 d可见自噬小体出现,之后自噬表达逐渐下降,细胞发生迟发性神经元死亡; QRHY方能减轻上述损害; ②I/R组LC3 mRNA及其蛋白表达均于缺血再灌注后12h开始明显上升,24 h达高峰(P<0.01),随后其表达渐趋下降(P<0.05); 而P62 mRNA及其蛋白表达时相变化则与之相反(P<0.05或P<0.01); QKL组较I/R组P62表达明显升高,LC3表达明显降低(P<0.05); QRHY 组较QKL组进一步升高P62表达,降低LC3的表达(P<0.05或P<0.01)。结论:清热化瘀方可能通过调节P62/LC3信号通路而减轻自噬的程度,从而达到减轻脑缺血再灌注损伤,保护神经元的作用。
Abstract:
Objective:To observe the protective effects of Qingre Huayu prescription against cerebral ischemia reperfusion(I/R)injury and relationship with the autophagy-related protein P62 and LC3 in rats.Method:160 rats were randomly divided into four groups:sham operation group(SO),model group(I/R),QRHY group and QKL control group.Each group was further divided into 4 subgroups according to 12 h,1 d,2 d and 3 d after I/R.The model of focal cerebral ischemia was prepared using Longa's method.The morphological changes of neuronal autophagy and apoptosis observed by transmission electron microscope,and the expression changes of P62 and LC3 mRNA and their protein observed by qRT-PCR and western blot methods separately.Results:①The autophagy was activated and the autophagosome was observed from 12 h to 3 d after I/R injury in rats.Compared with I/R group,the autophagy and apoptosis of QRHY group were significantly reduced.②The expression of LC3 mRNA and its protein in I/R group increased significantly after I/R,and 24h reached the peak(P<0.01),then decreased continuously,but still maintaining high expression levels.But the expression change of P62 mRNA and its protein were just the opposite at each point(P<0.05 or P<0.01).QKL could increase the expression of P62 and decrease the expression of LC3(P<0.05).QRHY could further induce the expression of P62 and decrease the expression of LC3 than that of QKL group(P<0.05).Conclusion:Qingre Huayu prescription could protect neurons and reduce the degree of autophagy through regulating the P62/LC3 signaling pathway after I/R in rats.

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备注/Memo

备注/Memo:
*国家自然科学基金资助项目(81560759)
更新日期/Last Update: 2020-04-13