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电针足三里对脓毒症模型大鼠肠道Ghrelin及高迁移率族蛋白B1表达的影响 被引量:13

Effect of Electro-acupuncture at Zusanli (ST36) on the Expression of Ghrelin and HMGB1 in the Small Intestine of Sepsis Rats
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摘要 目的探讨脓毒症模型大鼠血清及肠组织Ghrelin及高迁移率族蛋白B1(high mobility group protein,HMGB1)表达及电针足三里干预对其表达的影响。方法采用随机数字表法将48只Wistar雄性大鼠分为假手术组(Sham组)、盲肠结扎穿孔术组(cecal ligaton and puncture,CLP组)、CLP加电针足三里组(electroacupanctrue,EA组)及CLP加Ghrelin受体阻断剂加电针组(GHSRA组),每组12只。通过CLP术诱导制备严重腹腔感染致脓毒症模型。Sham组沿腹正中线切口后立即缝合腹壁切口。EA组于CLP术后20 min行持续针刺双侧足三里30 min(强度为2 mA,2~100 Hz),GHSRA组于电针前静脉注射Ghrelin受体阻断剂[D-Arg1,D-Phe5,D-Trp7^(7.9),Leu^(11)]-substance P,用量为700 nmol/kg。注射开始20 min后行电针足三里治疗,操作同EA组。各组大鼠于CLP术后12 h取血标本,采用E LISA法测定血清HMGB1及Ghrelin含量,免疫组化法测定肠组织Ghrelin免疫阳性细胞数目,蛋白质印迹法(Western blot)检测肠组织HMGB1蛋白表达。结果与Sham组比较,CLP组血清HMGB1水平及肠组织HMGB1蛋白表达显著增加(P<0.05),Ghrelin水平及其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HMGB1水平及肠组织HMGB1蛋白表达显著降低(P<0.05),而Ghrelin水平及其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HMGB1水平及肠组织HMGB1蛋白表达增加(P<O.05),但Ghrelin水平比较,差异无统计学意义(P>0.05);Sham、CLP和EA组大鼠血清HMGB1与Ghrelin水平呈负相关(r=-0.528,P<0.01)。结论电针足三里可抑制脓毒症模型大鼠肠道HMGB1蛋白表达,促进Ghrelin表达。Ghrelin受体阻断剂能阻滞电针对HMGB1的抑制作用,电针足三里的抗炎作用可能与Ghrelin有关。 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,[DArg^1,D-Phe^5,D-Trp^7.9,Leu^11]-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CLP,and 20 min later,EA at Zusanli(ST36) was performed in the same way as for rats in the EA group.Blood samples were withdrawn 12 h after CLP.The serum levels of Ghrelin and HMGB1 were detected using ELISA.Ghrelin expressions and the number of Ghrelin immunopositive cell in the jejunum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(P0.05).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.
出处 《中国中西医结合杂志》 CAS CSCD 北大核心 2014年第9期1113-1117,共5页 Chinese Journal of Integrated Traditional and Western Medicine
基金 浙江省中医药重大攻关项目(No.2012ZGG001)
关键词 脓毒症 电针 足三里 GHRELIN 高迁移率族蛋白B1 sepsis electro-acupuncture Zusanli(ST36) Ghrelin high mobility group boxl
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