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| 电刺激足三里穴增加miR-374表达促进自噬缓解肝缺血再灌注损伤机制研究 |
| 李叶晟,杨宗国,刘娇,陆云飞,陈晹,张猛,陈晓蓉,黄杨卿 |
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| 上海市公共卫生临床中心肝胆外科上海 201508;复旦大学附属肿瘤医院肝脏外科上海 200032;上海市公共卫生临床中心中医科上海 201508 |
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| 摘要: |
| 目的 探索电刺激足三里穴对肝缺血再灌注损伤的保护作用机制和原理。方法:将32只SD大鼠随机分为4组:假手术组、缺血再灌注组、足三里穴电刺激组、电激加miR-374阻断组，于再灌注8 h取材，每组8只。检测血清谷丙转氨酶(ALT)、肿瘤坏死因子(TNF)-α浓度，采用实时荧光定量PCR技术检测miR-374的水平。Western blot检测LC-3II、Beclin-1的蛋白水平。HE染色检测各实验组肝脏组织损伤情况。比较各组的上述因子水平。结果:与假手术组相比，缺血再灌注组血清ALT、TNF-α增加(P<0.001)，并随着miR-374表达的降低，LC-3II、Beclin-1表达的增加(P <0.01)，肝细胞损伤严重。与缺血组相比，足三里穴电刺激组的ALT、TNF-α下降(P <0.01)，同时miR-374上调，LC-3II、Beclin-1进一步增加(P <0.01)，HE染色发现肝损伤得到缓解。相较于足三里穴电刺激组，电激加miR-374阻断组逆转了电刺激的治疗作用(P <0.05)。结论:电刺激足三里穴能增加miR-374的表达，从而促进自噬，以减轻缺血再灌注诱导的肝脏损伤。 |
| 关键词: 肝脏缺血再灌注 电刺激穴位 miR-374 自噬 |
| DOI：10.3969/j.issn.1007-6948.2023.05.020 |
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| 基金项目:2020年上海市公共卫生临床中心院级科研课题(KY-GW-2020-06) |
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| Study on the mechanism of electrostimulation at Zusanli Point to increase miR-374 expression, promote autophagy and relieve hepatic ischemia reperfusion injury |
| Li Ye-sheng，Yang Zong-guo，Liu Jiao |
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| Abstract: |
| Objective To explore the protective mechanism and principle of Electro-Acupuncture at Zusanli (ST36) on liver ischemia-reperfusion injury. Methods Thirty-two rats were randomly divided into 4 groups: sham-operated group (NT), ischemia-reperfusion group (IR), electro-Acupuncture group (IR+EA), electro-acupuncture group+miR-374 inhibitor group (IR+EA+siRNA), electro-acupuncture group+miR-374 inhibitor group (IR+EA+siRNA). Samples were collected at 8 hours and each group has 8 rats. Firstly, the serum ALT level had been tested. The quantitative PCR was used to detected the expression level of miR-374. The Western blot was used to evaluate LC-3II and Beclin-1 protein level. The HE staining was used to measure the liver damage. The one-way ANOVA was used for comparison among multiple groups. Results Compared with sham-operated group, ischemia-reperfusion group had higher level of liver damage, serum ALT, TNF-α with lower expression of miR-374 and higher expression of LC-3II and Beclin-1. Compared with ischemia-reperfusion group, electro-acupuncture group had apparently decreased the liver damage, ALT, TNF-α with upregulated miR-374 and higher expression of LC-3II and Beclin-1. Compared with electro-acupuncture group, electro-acupuncture group+miR-374 inhibitor group reversed the therapeutical effect of electro-acupuncture. Conclusion Electro-acupuncture at Zusanli (ST36) upregulated the expression of miR-374, then promoted autophagy to mitigate the liver injury induced by ischemia-reperfusion. |
| Key words: Liver ischemia-reperfusion electro-acupuncture miR-374 autophagy |
