[电针足三里（ST36）对脓毒症大鼠小肠中胃饥饿素和高迁移率族蛋白B1表达的影响]

[Effect of electro-acupuncture at zusanli (ST36) on the expression of ghrelin and HMGB1 in the small intestine of sepsis rats].

作者信息

Wu Jian-Nong, Wu Wan, Jiang Rong-Lin, Zhu Mei-Fei, Lei Shu, Lu Bin

出版信息

Zhongguo Zhong Xi Yi Jie He Za Zhi. 2014 Sep;34(9):1113-7.

Abstract

OBJECTIVE

To explore the expression of Ghrelin and high mobility group protein B1 (HMGB1) in the serum and the intestinal tissue of sepsis model rats, and to evaluate the effect of electro-acupuncture (EA) at Zusanli (ST36) on the expression of HMGB1 and Ghrelin.

METHODS

Forty-eight male Wistar rats were randomly divided into four groups, i.e., the sham-operation (sham), the cecal ligation and puncture group (CLP), the CLP + EA at Zusanli (ST36) group (EA), and the CLP + Ghrelin receptor blocking agent + EA group (GHSRA), 12 in each group. A sepsis rat model was prepared by CLP. The incision of the abdominal wall was immediately sutured along the ventral midline for rats in the Sham group. In the EA group EA at Zusanli (ST36) was performed 20 min after CLP surgery with the constant voltage (2 - 100 Hz, 2 mA) for 30 min. In the GHSRA group, Ghrelin receptor blocking agent, [D-Arg1, D-Phe5, D-Trp79, Leu11]-substance P (700 nmol/kg), was administered through intravenous injection immediately after CLP, and 20 min later, EA at Zusanli (ST36) was performed in the same way as for rats in the EA group. Blood samples were withdrawn 12 h after CLP. The serum levels of Ghrelin and HMGB1 were detected using ELISA. Ghrelin expressions and the number of Ghrelin immunopositive cell in the jejunum were determined by immunohistochemistry. HMGB1 contents of the jejunum tissue were detected by Western blotting.

RESULTS

Compared with the Sham group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly increased and levels of Ghrelin and the expression rate of immunopositive cells significantly decreased in the CLP group (P < 0.05). Compared with the CLP group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly decreased, but levels of Ghrelin and the expression rate of immunopositive cells significantly increased in the EA group (P < 0.05). Compared with the EA group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly increased in the GHSRA group (P < 0.05), but there was no statistical difference in levels of Ghrelin between the two groups (P > 0.05). The serum level of HMGB1 was negatively correlated with Ghrelin in the Sham group, the CLP group, and the EA group (r = -0. 528, P < 0.01).

CONCLUSIONS

EA at Zusanli (ST36) could inhibit the expression of HMGB1 in the jejunum of septic rats, and promote the expression of Ghrelin. The expression of HMGB1 was inhibited by Ghrelin receptor blocking agent, which suggested that the anti-inflammation of EA at Zusanli (ST36) might be associated with Ghrelin.

摘要

目的

探讨脓毒症模型大鼠血清及肠组织中胃饥饿素（Ghrelin）和高迁移率族蛋白B1（HMGB1）的表达，评价足三里（ST36）电针（EA）对HMGB1和Ghrelin表达的影响。

方法

48只雄性Wistar大鼠随机分为四组，即假手术组（sham）、盲肠结扎穿孔组（CLP）、CLP + 足三里（ST36）电针组（EA）、CLP + 胃饥饿素受体阻断剂 + 电针组（GHSRA），每组12只。采用CLP制备脓毒症大鼠模型。假手术组大鼠沿腹中线立即缝合腹壁切口。EA组在CLP术后20 min进行足三里（ST36）电针，恒压（2 - 100 Hz，2 mA），持续30 min。GHSRA组在CLP后立即静脉注射胃饥饿素受体阻断剂[D-Arg1，D-Phe5，D-Trp79，Leu11]-P物质（700 nmol/kg），20 min后，以与EA组大鼠相同的方式进行足三里（ST36）电针。CLP术后12 h采集血样。采用酶联免疫吸附测定法（ELISA）检测血清中Ghrelin和HMGB1水平。通过免疫组织化学法测定空肠中Ghrelin表达及Ghrelin免疫阳性细胞数量。采用蛋白质免疫印迹法检测空肠组织中HMGB1含量。

结果

与假手术组比较，CLP组血清免疫阳性细胞数量及空肠组织中HMGB1表达显著增加，Ghrelin水平及免疫阳性细胞表达率显著降低（P < 0.05）。与CLP组比较，EA组血清免疫阳性细胞数量及空肠组织中HMGB1表达显著降低，但Ghrelin水平及免疫阳性细胞表达率显著增加（P < 0.05）。与EA组比较，GHSRA组血清免疫阳性细胞数量及空肠组织中HMGB1表达显著增加（P < 0.05），但两组间Ghrelin水平差异无统计学意义（P > 0.05）。假手术组、CLP组和EA组血清HMGB1水平与Ghrelin呈负相关（r = -0. 528，P < 0.01）。