电针“足三里”穴对脾气虚大鼠小肠黏膜转运体SGLT1及GLUT2表达的影响
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摘要
目的:观察电针双侧"足三里"穴对脾气虚模型大鼠小肠黏膜上皮组织内的钠依赖葡萄糖转运体(SGLT1),葡萄糖运载蛋白2(GLUT2)的基因及蛋白表达的影响。方法:将40只SD雄性大鼠随机分为正常组、脾气虚组、足三里组、非经非穴组,每组10只。所有大鼠均在SPF级动物实验中心饲养。除正常组外,其余3组均采用劳倦过度和不规则饮食复合法建立脾气虚证的大鼠模型。造模成功之后,对足三里组、非经非穴组大鼠分别进行电针"足三里"穴、非经非穴点干预处理7天。采用HE染色方法观察各组大鼠小肠黏膜组织形态变化;采用荧光定量PCR法检测大鼠小肠黏膜上皮的SGLT1和GLUT2的mRNA表达水平;采用蛋白质免疫印迹法(WB)检测大鼠小肠黏膜上皮组织内的SGLT1和GLUT2的蛋白含量变化。结果:脾气虚组大鼠小肠黏膜组织部分损伤,足三里组的小肠黏膜组织有一定程度的恢复;脾气虚组和非经非穴组大鼠小肠黏膜组织内的SGLT1和GLUT2蛋白和基因表达水平均明显低于正常组(P <0. 05);足三里组大鼠小肠黏膜组织内的SGLT1和GLUT2蛋白和基因表达水平相比于脾气虚组有所升高(P <0. 05),非经非穴组未见显著性差异(P> 0. 05)。结论:电针"足三里"穴可以调节脾气虚大鼠小肠黏膜上皮组织内的SGLT1和GLUT2基因及蛋白的异常表达,参与小肠对葡萄糖的吸收作用进而改善脾气虚证。
Objective: To observe the effect of electroacupuncture at Zusanli( ST36) on the gene and protein expression of sodium dependent glucose transporter( SGLT1) and glucose carrier protein( GLUT2) in the epithelium of small intestinal mucosa of rats with spleen Qi deficiency. Methods: Forty SD rats were divided into two groups. Male rats were randomly divided into normal group,spleen Qi deficiency group,Zusanli( ST36) group,non-meridian non-acupoint group,10 rats in each group. All rats were raised in SPF level animal experimental center. The other three groups were used to establish the rat model of spleen Qi deficiency by the method of overfatigue and irregular diet. After the model was established,the rats in Zusanli( ST36) group and non-meridian non-acupoint group were treated respectively with Zusanli( ST36) group and non-meridian non-acupoint group. Electroacupuncture at Zusanli( ST36) was performed for 7 days without intervention at non-acupoint point. The morphological changes of small intestine mucosa were observed by HE staining method,and the expressions of SGLT1 and GLUT2 in rat small intestinal mucosa epithelium were detected by fluorescence quantitative PCR method. The protein contents of SGLT1 and GLUT2 in small intestinal mucosa of rats were detected by Western blot. Results: The intestinal mucosal tissue was partially damaged in spleen Qi deficiency group and recovered to some extent in Zusanli( ST36) group. The levels of SGLT1 and GLUT2 protein and gene expression in small intestinal mucosa of rats with deficiency of spleen Qi and non-acupoints were significantly lower than those of normal group( P < 0. 05),and the expression levels of SGLT1 and GLUT2 protein and gene in small intestinal mucosa of Zusanli( ST36) group were significantly lower than those of the normal group( P < 0. 05),and those of Zusanli( ST36) group were significantly lower than those of the normal group( P < 0. 05). There was no significant difference in spleen Qi deficiency group( P <0. 05),but no significant difference was found in non-meridian non-point group( P < 0. 05). Conclusion: Electroacupuncture at Zusanli( ST36) can regulate the abnormal expressions of SGLT1 and GLUT2 gene and protein in the epithelium of small intestine mucosa of rats with spleen Qi deficiency,and participate in the absorption of glucose in the small intestine and then improve the spleen Qi deficiency syndrome.
Objective: To observe the effect of electroacupuncture at Zusanli( ST36) on the gene and protein expression of sodium dependent glucose transporter( SGLT1) and glucose carrier protein( GLUT2) in the epithelium of small intestinal mucosa of rats with spleen Qi deficiency. Methods: Forty SD rats were divided into two groups. Male rats were randomly divided into normal group,spleen Qi deficiency group,Zusanli( ST36) group,non-meridian non-acupoint group,10 rats in each group. All rats were raised in SPF level animal experimental center. The other three groups were used to establish the rat model of spleen Qi deficiency by the method of overfatigue and irregular diet. After the model was established,the rats in Zusanli( ST36) group and non-meridian non-acupoint group were treated respectively with Zusanli( ST36) group and non-meridian non-acupoint group. Electroacupuncture at Zusanli( ST36) was performed for 7 days without intervention at non-acupoint point. The morphological changes of small intestine mucosa were observed by HE staining method,and the expressions of SGLT1 and GLUT2 in rat small intestinal mucosa epithelium were detected by fluorescence quantitative PCR method. The protein contents of SGLT1 and GLUT2 in small intestinal mucosa of rats were detected by Western blot. Results: The intestinal mucosal tissue was partially damaged in spleen Qi deficiency group and recovered to some extent in Zusanli( ST36) group. The levels of SGLT1 and GLUT2 protein and gene expression in small intestinal mucosa of rats with deficiency of spleen Qi and non-acupoints were significantly lower than those of normal group( P < 0. 05),and the expression levels of SGLT1 and GLUT2 protein and gene in small intestinal mucosa of Zusanli( ST36) group were significantly lower than those of the normal group( P < 0. 05),and those of Zusanli( ST36) group were significantly lower than those of the normal group( P < 0. 05). There was no significant difference in spleen Qi deficiency group( P <0. 05),but no significant difference was found in non-meridian non-point group( P < 0. 05). Conclusion: Electroacupuncture at Zusanli( ST36) can regulate the abnormal expressions of SGLT1 and GLUT2 gene and protein in the epithelium of small intestine mucosa of rats with spleen Qi deficiency,and participate in the absorption of glucose in the small intestine and then improve the spleen Qi deficiency syndrome.
引文
[1]Dong R,Srai SK,Debnam E,et al. Transcriptional and translational control over sodium-glucose linked transporter(SGLT1)gene expression in adult rat small intestine[J]. FEBS Lett,1997,406(122):792-782.
[2]施旭光,施家希,刘海涛,等.补中益气汤对脾虚泄泻大鼠SGLT1/NHE3通路的影响[J].中药新药与临床药理,2018,29(1):8-12.
[3]刘海涛,施家希,黄张杰,等.补中益气汤不同配伍对脾虚大鼠免疫器官及小肠黏膜转运体蛋白表达的影响[J].中药材,2017,40(2):466-469.
[4]陈婷.基于SGLI1通路探讨四君子汤对Caco-2细胞葡萄糖吸收的影响[D].广州:广州中医药大学,2016.
[5]郑小伟.中医实验动物模型方法学[M].上海:上海中医药大学出版社,1999:63-111.
[6]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:327.
[7]林莺,纪峰,黄桂榕,等.针刺实验研究中大鼠非经非穴选取方法及思考[J].针刺研究,2013,38(4):334-338.
[8]刘沐苍.针刺足三里穴治疗功能性消化不良大鼠实验研究[D].大连:大连医科大学,2016.
[9]韩春晓,王江,车艳秋,等.针刺足三里的脊髓背根神经电信号非线性特征提取[J].物理学报,2010,59(8):5881-5888.
[10]高巍,黄裕新,陈洪,等.电针“足三里”对大鼠脑肠肽含量的影响及其对免疫系统的调控作用[J].针刺研究,2002,27(1):50-55.
[11]Kellett GL,Brot-Laroche E,Mace OJ,et al. Sugar absorp-tion in the intestine:the role of GLUT2[J]. Annu Rev Nu-tr,2008,28(S1):35-54.
[12]Stolarczyk E,Le Gall M,Even P,et al. Loss of sugar detec-tion by GLUT2 affects glucose homeostasis in mice[J]. PLo S One,2007,2(12):e1288.
[13]Lostao MP,Berjon A,Barber A,et al. On the multiplicity of glucose analogues transport systems in rat intestine[J]. Revista Espanola de Fisiologia,1991,47:209-216.
[14]Dyer J,Dalv K,Salmon KS,et al. Intestinal glucose sensing and regulation of intestinal glucose absorption[J]. BiochemSoc Trans,2007,35:1191-1194.
[15]郑媛嘉,张磊,郭文峰.黄芪对脾虚大鼠小肠葡萄糖吸收功能的影响及机制[J].中药新药与临床药理,2017,28(6):703-707.
[16]吴燕燕,黄丽花,王晶,等.精眯对脾虚大鼠肠黏膜损伤的修复及葡萄糖吸收的影响[J].广东药科大学学报,2017,33(1):88-93.
[17]张磊.黄芪对肠上皮细胞葡萄糖吸收的影响及其调控机制的研究[D].广州:广州中医药大学,2016.
[18]黄丽花.四君子汤对脾虚大鼠肠黏膜修复及葡萄糖吸收的机理研究[D].广州:广东药科大学,2016.
[19]黄张杰.补中益气汤及其“益气升阳”配比组方的药效和代谢组学研究[D].广州:广州中医药大学,2014.
[20]王桂香,黎同明,王素军,等.参苓白术散对脾虚泄泻大鼠SGLT1mRNA表达的影响[J].中药材,2013,36(7):1140-1143.
[2]施旭光,施家希,刘海涛,等.补中益气汤对脾虚泄泻大鼠SGLT1/NHE3通路的影响[J].中药新药与临床药理,2018,29(1):8-12.
[3]刘海涛,施家希,黄张杰,等.补中益气汤不同配伍对脾虚大鼠免疫器官及小肠黏膜转运体蛋白表达的影响[J].中药材,2017,40(2):466-469.
[4]陈婷.基于SGLI1通路探讨四君子汤对Caco-2细胞葡萄糖吸收的影响[D].广州:广州中医药大学,2016.
[5]郑小伟.中医实验动物模型方法学[M].上海:上海中医药大学出版社,1999:63-111.
[6]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:327.
[7]林莺,纪峰,黄桂榕,等.针刺实验研究中大鼠非经非穴选取方法及思考[J].针刺研究,2013,38(4):334-338.
[8]刘沐苍.针刺足三里穴治疗功能性消化不良大鼠实验研究[D].大连:大连医科大学,2016.
[9]韩春晓,王江,车艳秋,等.针刺足三里的脊髓背根神经电信号非线性特征提取[J].物理学报,2010,59(8):5881-5888.
[10]高巍,黄裕新,陈洪,等.电针“足三里”对大鼠脑肠肽含量的影响及其对免疫系统的调控作用[J].针刺研究,2002,27(1):50-55.
[11]Kellett GL,Brot-Laroche E,Mace OJ,et al. Sugar absorp-tion in the intestine:the role of GLUT2[J]. Annu Rev Nu-tr,2008,28(S1):35-54.
[12]Stolarczyk E,Le Gall M,Even P,et al. Loss of sugar detec-tion by GLUT2 affects glucose homeostasis in mice[J]. PLo S One,2007,2(12):e1288.
[13]Lostao MP,Berjon A,Barber A,et al. On the multiplicity of glucose analogues transport systems in rat intestine[J]. Revista Espanola de Fisiologia,1991,47:209-216.
[14]Dyer J,Dalv K,Salmon KS,et al. Intestinal glucose sensing and regulation of intestinal glucose absorption[J]. BiochemSoc Trans,2007,35:1191-1194.
[15]郑媛嘉,张磊,郭文峰.黄芪对脾虚大鼠小肠葡萄糖吸收功能的影响及机制[J].中药新药与临床药理,2017,28(6):703-707.
[16]吴燕燕,黄丽花,王晶,等.精眯对脾虚大鼠肠黏膜损伤的修复及葡萄糖吸收的影响[J].广东药科大学学报,2017,33(1):88-93.
[17]张磊.黄芪对肠上皮细胞葡萄糖吸收的影响及其调控机制的研究[D].广州:广州中医药大学,2016.
[18]黄丽花.四君子汤对脾虚大鼠肠黏膜修复及葡萄糖吸收的机理研究[D].广州:广东药科大学,2016.
[19]黄张杰.补中益气汤及其“益气升阳”配比组方的药效和代谢组学研究[D].广州:广州中医药大学,2014.
[20]王桂香,黎同明,王素军,等.参苓白术散对脾虚泄泻大鼠SGLT1mRNA表达的影响[J].中药材,2013,36(7):1140-1143.