[电针“肝俞”（BL18）和“阳陵泉”（GB34）通过抑制大鼠高迁移率族蛋白1的转位和释放减轻肝脏缺血再灌注损伤]

[Electroacupuncture at "Ganshu" (BL18) and "Yanglingquan" (GB34) alleviates hepatic ischemia-reperfusion injury by inhibiting translocation and release of high mobility group protein 1 in rats].

作者信息

Tan Si-You, Huang Jun, Su Ying-Ying, Chen Wen-Yan, Kong Gao-Yin, Liu Jing-Shi, Wei Lai

机构信息

Department of Anesthesiology, Hunan Provincial People's Hospital/The First Affiliated Hospital of Hunan Normal University, Changsha 410002, China.

Department of Anesthesiology, Hunan Provincial People's Hospital/The First Affiliated Hospital of Hunan Normal University, Changsha 410002, China; Hunan Province Perioperative Accelerated Rehabilitation Anesthesia Clinical Medical Research Center, Changsha 410002.

出版信息

Zhen Ci Yan Jiu. 2020 Nov 25;45(11):888-94. doi: 10.13702/j.1000-0607.200374.

DOI:10.13702/j.1000-0607.200374

PMID:33269832

Abstract

OBJECTIVE

To explore the protective effect of electroacupuncture (EA) on hepatic ischemia-reperfusion injury (HIRI) and the expression of high mobility group protein 1 (HMGB1) in liver tissues in rats.

METHODS

A total of 40 male SD rats were randomly divided into 4 groups, namely sham control, HIRI model, "Ganshu"(BL18) -"Yanglingquan"(GB34) and non-acupoint group, with 10 rats in each group. The HIRI model was induced by blocking the arteries, veins and bile ducts supplying the middle and left lobes of the liver for 1 h, and reperfusion for 4 h to induce an area of about 70% HIRI. EA was applied to bila-teral BL18 and GB34, or non-acupoints about 6-8 mm to the bilateral BL18 for 30 min before modeling. Serum alanine transaminase (ALT) and aspartate aminotransferase (AST) levels were measured by using an automatic biochemical analyzer. Serum tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and HMGB1 levels were assayed by ELISA. Hematoxylin - eosin (H.E.) staining was used to observe histopathological changes of the liver tissue by using tissue injury scaling (0-3 scores). The expression of HMGB1 protein in liver tissues was detected by immunohistochemical staining, Western blot and PCR, separately.

RESULTS

Following modeling and compared with the sham group, the levels of serum ALT, AST, TNF-α, IL-6, and HMGB1 contents, the number of HMGB1 immunoreaction (IR)-positive cells, and HMGB1 protein and mRNA were significantly increased (P＜0.01). After the treatment, the contents of serum ALT, AST, TNF-α, IL-6, and HMGB1, liver HMGB1 IR-positive cells, protein and mRNA were considerably down-regulated in the BL18-GB34 group (<0.05), rather than in the non-acupoint group (>0.05) in contrast to the model group. H.E. stain showed a higher liver injury score in the model group than in the sham group (<0.01), and a lower liver injury score in the BL18-GB34 group (not the non-acupoint group) relevant to the model group (<0.05).

CONCLUSION

EA of BL18 and GB34 points has a protective effect on ischemic liver injury in rats with HIRI, which may be associated with its functions in inhibiting the migration and release of HMGB1 from the nucleus to the cytoplasm and in down-regulating the expression of inflammatory factors.

摘要

目的

探讨电针（EA）对大鼠肝缺血再灌注损伤（HIRI）的保护作用及肝组织中高迁移率族蛋白1（HMGB1）的表达。

方法

将40只雄性SD大鼠随机分为4组，即假手术对照组、HIRI模型组、“肝俞”（BL18）-“阳陵泉”（GB34）组和非穴位组，每组10只。通过阻断供应肝中叶和左叶的动脉、静脉和胆管1小时，再灌注4小时诱导约70%的HIRI区域，建立HIRI模型。在建模前，将电针分别应用于双侧BL18和GB34，或双侧BL18旁约6 - 8 mm的非穴位，持续30分钟。使用自动生化分析仪测定血清丙氨酸转氨酶（ALT）和天冬氨酸转氨酶（AST）水平。采用酶联免疫吸附测定法（ELISA）检测血清肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和HMGB1水平。采用苏木精-伊红（H.E.）染色，通过组织损伤评分（0 - 3分）观察肝组织的组织病理学变化。分别采用免疫组织化学染色、蛋白质免疫印迹法（Western blot）和聚合酶链反应（PCR）检测肝组织中HMGB1蛋白的表达。

结果

建模后与假手术组相比，血清ALT、AST、TNF-α、IL-6水平及HMGB1含量、HMGB1免疫反应（IR）阳性细胞数、HMGB1蛋白和mRNA均显著升高（P＜0.01）。治疗后，与模型组相比，BL18 - GB34组血清ALT、AST、TNF-α、IL-6及HMGB1含量、肝脏HMGB1 IR阳性细胞、蛋白和mRNA均明显下调（P＜0.05），而非穴位组无明显变化（P＞0.05）。H.E.染色显示，模型组肝损伤评分高于假手术组（P＜0.01），与模型组相比，BL18 - GB34组（而非非穴位组）肝损伤评分降低（P＜0.05）。