不同灸量隔姜灸对脾虚证大鼠三叶因子1、粘蛋白5AC及表皮生长因子受体的影响（英文）
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摘要
目的:观察不同灸量隔姜灸对脾虚证大鼠血清三叶因子1(TFF1)和粘蛋白5AC(MUC5AC)含量,以及胃黏膜表皮生长因子受体(EGFR)蛋白表达的影响,探讨隔姜灸治疗脾虚证的可能作用机制及量效特征。方法:将75只SPF级Sprague-Dawley(SD)大鼠按随机数字表法分为空白对照组(A组)、模型组(B组)、隔姜灸3壮组(C1组)、隔姜灸6壮组(C2组)和隔姜灸9壮组(C3组),每组15只。除A组外,其余各组大鼠采用200%的大黄浓缩液4℃灌胃制作脾虚证大鼠模型。造模成功后,B组大鼠不予治疗;C1、C2和C3组大鼠分别接受3壮、6壮和9壮隔姜灸足三里和中脘治疗,连续治疗8 d。观察大鼠一般症状评分,采用酶联免疫吸附法(ELISA)检测血清中TFF1和MUC5AC含量;免疫组化法检测胃黏膜EGFR蛋白表达。结果:干预结束后,与A组比较,B组大鼠脾虚症状积分增高,C1、C2和C3组大鼠血清TFF1、MUC5AC含量及胃组织EGFR蛋白表达明显升高(均P<0.01);与B组比较,C1、C2和C3组大鼠脾虚症状积分降低,血清TFF1、MUC5AC含量及胃组织EGFR蛋白表达升高(均P<0.01);与C1组比较,C2和C3组大鼠脾虚症状积分降低,血清TFF1、MUC5AC含量及胃组织EGFR蛋白表达升高(均P<0.01),但C2组与C3组差异无统计学意义(均P>0.05)。结论:隔姜灸能改善大鼠脾虚症状,促进脾虚证大鼠胃黏膜的增殖修复,其作用机制可能与提高血清TFF1和MUC5AC含量,激活EGFR蛋白的表达相关,且灸9壮和6壮的疗效优于灸3壮,但灸9壮和灸6壮的效果相当。
Objective: To observe the effects of different doses of ginger-partitioned moxibustion on serum trefoil factor 1(TFF1) and mucin 5AC(MUC5AC) levels, as well as the expression of epidermal growth factor receptor(EGFR) in gastric mucosa of rats with spleen deficiency syndrome, therefore, to explore the possible mechanism and the dose-effect characteristics of ginger-partitioned moxibustion in spleen deficiency syndrome.Methods: Seventy-five SPF grade Sprague-Dawley(SD) rats were randomly divided into a blank control group(group A), a model group(group B), a 3 moxa-cone ginger-partitioned moxibustion group(group C1), a 6 moxa-cone ginger-partitioned moxibustion group(group C2) and a 9 moxa-cone ginger-partitioned moxibustion group(group C3) using random number table method, 15 rats in each group. Except group A, rats in the other groups received intragastric administration of 4 ℃200% concentrated Da Huang(Radix et Rhizoma Rhei) to prepare spleen deficiency syndrome model. After successful modeling, rats in group B received no treatment; rats in group C1, C2 and C3 were treated with 3, 6 and 9 moxa-cone ginger-partitioned moxibustion at Zusanli(ST 36) and Zhongwan(CV 12) respectively for 8 continuous days. The general symptom score of rats was observed. The serum levels of TFF1 and MUC5 AC were detected by enzyme-linked immunosorbent assay(ELISA). The expression of EGFR protein in gastric mucosa was detected by immunohistochemistry.Results: After the treatment, compared with group A, the spleen deficiency symptom score was increased in group B, the levels of serum TFF1 and MUC5AC, the EGFR protein expression in gastric tissues of group C1, C2 and C3 were significantly increased(all P<0.01); compared with group B, the spleen deficiency scores were decreased in group C1, C2 and C3, and the serum levels of TFF1 and MUC5AC, as well as EGFR protein expression in gastric tissues were increased(all P<0.01).Compared with group C1, the spleen deficiency scores were decreased in group C2 and C3, the serum levels of TFF1 and MUC5AC, and the expression of EGFR protein in gastric tissues were increased(all P<0.01), however, there was no significant difference between group C2 and C3(all P>0.05). The mechanism may be related to the increase of serum TFF1 and MUC5AC levels and activation of EGFR protein.Conclusion: Ginger-partitioned moxibustion can improve the symptoms, as well as promote the proliferation and repair of gastric mucosa in rats with spleen deficiency. The therapeutic efficacy of 6 or 9 moxa-cone ginger-partitioned moxibustion is better than that of 3 moxa-cone ginger-partitioned moxibustion, while the efficacies are equivalent between 6 and 9 moxa-cone ginger-partitioned moxibustion groups.
Objective: To observe the effects of different doses of ginger-partitioned moxibustion on serum trefoil factor 1(TFF1) and mucin 5AC(MUC5AC) levels, as well as the expression of epidermal growth factor receptor(EGFR) in gastric mucosa of rats with spleen deficiency syndrome, therefore, to explore the possible mechanism and the dose-effect characteristics of ginger-partitioned moxibustion in spleen deficiency syndrome.Methods: Seventy-five SPF grade Sprague-Dawley(SD) rats were randomly divided into a blank control group(group A), a model group(group B), a 3 moxa-cone ginger-partitioned moxibustion group(group C1), a 6 moxa-cone ginger-partitioned moxibustion group(group C2) and a 9 moxa-cone ginger-partitioned moxibustion group(group C3) using random number table method, 15 rats in each group. Except group A, rats in the other groups received intragastric administration of 4 ℃200% concentrated Da Huang(Radix et Rhizoma Rhei) to prepare spleen deficiency syndrome model. After successful modeling, rats in group B received no treatment; rats in group C1, C2 and C3 were treated with 3, 6 and 9 moxa-cone ginger-partitioned moxibustion at Zusanli(ST 36) and Zhongwan(CV 12) respectively for 8 continuous days. The general symptom score of rats was observed. The serum levels of TFF1 and MUC5 AC were detected by enzyme-linked immunosorbent assay(ELISA). The expression of EGFR protein in gastric mucosa was detected by immunohistochemistry.Results: After the treatment, compared with group A, the spleen deficiency symptom score was increased in group B, the levels of serum TFF1 and MUC5AC, the EGFR protein expression in gastric tissues of group C1, C2 and C3 were significantly increased(all P<0.01); compared with group B, the spleen deficiency scores were decreased in group C1, C2 and C3, and the serum levels of TFF1 and MUC5AC, as well as EGFR protein expression in gastric tissues were increased(all P<0.01).Compared with group C1, the spleen deficiency scores were decreased in group C2 and C3, the serum levels of TFF1 and MUC5AC, and the expression of EGFR protein in gastric tissues were increased(all P<0.01), however, there was no significant difference between group C2 and C3(all P>0.05). The mechanism may be related to the increase of serum TFF1 and MUC5AC levels and activation of EGFR protein.Conclusion: Ginger-partitioned moxibustion can improve the symptoms, as well as promote the proliferation and repair of gastric mucosa in rats with spleen deficiency. The therapeutic efficacy of 6 or 9 moxa-cone ginger-partitioned moxibustion is better than that of 3 moxa-cone ginger-partitioned moxibustion, while the efficacies are equivalent between 6 and 9 moxa-cone ginger-partitioned moxibustion groups.
引文
[1]Duan YQ,Cheng WD,Cheng YX,Yang XY,Li LZ,An YR,Tian R,Wang Y.Investigation of syndrome mode connotation for the‘spleen deficiency syndrome body generalization effect’based on the principle of spleen deficiency syndrome.Zhongguo Zhongyiyao Xinxi Zazhi,2013,20(7):3-5.
[2]Cheng YM,Lu MT,Yeh CM.Functional expression of recombinant human trefoil factor 1 by Escherichia coli and Brevibacillus choshinensis.BMC Biotechnol,2015,15(1):1-13.
[3]Rao J,Yang XJ,Fan DM,Chen WQ,Wang JH.Effect the treatment of Jianpi Qingre Huayu decoction with gastric precancerous lesions and its influence on mucin 5ACprotein expression.Zhongguo Shiyan Fangjixue Zazhi,2014,20(4):183-187.
[4]Dahlhoff M,Gerhard M,Rad R,Lindén S,Wolf E,Schneider MR.A new mouse model for studying EGFR-dependent gastric polyps.Biochim Biophys Acta,2012,1822(8):1293-1299.
[5]Yi Z,Chang XR,Yi SX,Yan J,Xie H,Lin YP,Song J,Peng F.Evidence-based medicine analysis of clinical studies survey on the acupuncture-moxibustion treatment of spleen-stomach vacuity cold.Zhonghua Zhongyiyao Xuekan,2010,28(9):1866-1868.
[6]Yi Z,Liu M,Chang XR,Yan J,Xie H,Wang DJ,Ai K,Liu WA.Influence of serum SOD and MDA by moxibustion on ginger therapy for chronic superficial gastritis spleenstomach vacuity cold patients with different time.Zhongguo Shiyan Fangjixue Zazhi,2012,18(23):301-304.
[7]Peng Y,Peng F,Yi SX,Lin YP,Chang XR,Long YW,Zhang HG.Effect of moxibustion on motility,absorption and content of ATP in small intestine of spleen-deficiency rats.Zhongguo Zhen Jiu,2012,32(3):246-250.
[8]Li ZR.Experimental Acupuncture Science.2nd Edition.Beijing:China Press of Traditional Chinese Medicine,2007:253.
[9]Shi T,Zhu FS,Xu TT.The correlation analysis between spleen deficiency and gastric mucosal injury.Yixue Yu Zhexue(Linchuang Juece Luntan Ban),2011,32(12):59-60.
[10]Tan J,Chang XR,Yan J,Yi SX,Lin YP,Yue ZH,Liu M,Peng Y.Effect of moxibustion on plasma levels ofβ-EP,MTL and SS in rats with spleen deficiency.Shijie Huaren Xiaohua Zazhi,2011,19(34):3498-3502.
[11]Chang XR,Liu M,Yan J,Yi SX,Yue ZH,Zhang GS,Liu ML,Sun GJ,Wang LL,Hu L,Wu HG.Research on mechanisms and principles of warm-unblock and warmtonic effects on moxibustion.Shijie Zhongyiyao,2013,8(8):875-879.
[12]Sun Y,Wang LX.The research progress of trefoil factor family.Changwai Yu Changnei Yingyang,2015,22(1):49-52.
[13]Ren JL,Lu YP,Wang L,Chen JM,Shi HX,Ye ZS,Wu YH,Zhong Y,Lin XT,Lin H,Pan JS,Luo JY.Express change of TFF1 in normal and injured gastric mucosa.Shijie Huaren Xiaohua Zazhi,2003,11(11):1809-1810.
[14]Li TL,Su YM,Qi F,Ni W,Zhang H,Zhang YC,Guo H.Effect of moxibustion on serum TFF and gastric mucosa ERK1/2 and PCNA in gastric ulcer model rats with spleen deficiency symptom.Hunan Zhongyiyao Daxue Xuebao,2015,35(2):49-51.
[15]Song J,Guo RF,Su RL,Qin EP,Wang JL.Expressions and significance of TFF2,CLDN18 and MUC5AC protein in gastric mucosal lesions.Wei Chang Bing Xue,2015,20(7):394-397.
[16]Zhang H,Guo H,Zhang YC,Liu M,Ai K,Su YM,Li MH,Li TL.Effect of moxibustion intervention on expression of gastric epidermal growth factor receptor and extracellular signal regulated kinase 1/2 expression in rats with gastric ulcer.Zhen Ci Yan Jiu,2014,39(5):351-357.
[17]Liu XL,Wang RJ,Zhao QQ.Effect on expressions of MUC5AC mRNA and protein in gastric mucosa of Bu Zhong Yi Qi decoction for rats with spleen deficiency.Zhongyao Yaoli Yu Linchuang,2012,28(4):7-9.
[18]Liu XL,Wang RJ,Zhang QQ.Effect on EGFR protein expression in gastric mucosa of Bu Zhong Yi Qi decoction for rats with spleen deficiency.Zhongyao Yaoli Yu Linchuang,2012,28(5):12-14.
[2]Cheng YM,Lu MT,Yeh CM.Functional expression of recombinant human trefoil factor 1 by Escherichia coli and Brevibacillus choshinensis.BMC Biotechnol,2015,15(1):1-13.
[3]Rao J,Yang XJ,Fan DM,Chen WQ,Wang JH.Effect the treatment of Jianpi Qingre Huayu decoction with gastric precancerous lesions and its influence on mucin 5ACprotein expression.Zhongguo Shiyan Fangjixue Zazhi,2014,20(4):183-187.
[4]Dahlhoff M,Gerhard M,Rad R,Lindén S,Wolf E,Schneider MR.A new mouse model for studying EGFR-dependent gastric polyps.Biochim Biophys Acta,2012,1822(8):1293-1299.
[5]Yi Z,Chang XR,Yi SX,Yan J,Xie H,Lin YP,Song J,Peng F.Evidence-based medicine analysis of clinical studies survey on the acupuncture-moxibustion treatment of spleen-stomach vacuity cold.Zhonghua Zhongyiyao Xuekan,2010,28(9):1866-1868.
[6]Yi Z,Liu M,Chang XR,Yan J,Xie H,Wang DJ,Ai K,Liu WA.Influence of serum SOD and MDA by moxibustion on ginger therapy for chronic superficial gastritis spleenstomach vacuity cold patients with different time.Zhongguo Shiyan Fangjixue Zazhi,2012,18(23):301-304.
[7]Peng Y,Peng F,Yi SX,Lin YP,Chang XR,Long YW,Zhang HG.Effect of moxibustion on motility,absorption and content of ATP in small intestine of spleen-deficiency rats.Zhongguo Zhen Jiu,2012,32(3):246-250.
[8]Li ZR.Experimental Acupuncture Science.2nd Edition.Beijing:China Press of Traditional Chinese Medicine,2007:253.
[9]Shi T,Zhu FS,Xu TT.The correlation analysis between spleen deficiency and gastric mucosal injury.Yixue Yu Zhexue(Linchuang Juece Luntan Ban),2011,32(12):59-60.
[10]Tan J,Chang XR,Yan J,Yi SX,Lin YP,Yue ZH,Liu M,Peng Y.Effect of moxibustion on plasma levels ofβ-EP,MTL and SS in rats with spleen deficiency.Shijie Huaren Xiaohua Zazhi,2011,19(34):3498-3502.
[11]Chang XR,Liu M,Yan J,Yi SX,Yue ZH,Zhang GS,Liu ML,Sun GJ,Wang LL,Hu L,Wu HG.Research on mechanisms and principles of warm-unblock and warmtonic effects on moxibustion.Shijie Zhongyiyao,2013,8(8):875-879.
[12]Sun Y,Wang LX.The research progress of trefoil factor family.Changwai Yu Changnei Yingyang,2015,22(1):49-52.
[13]Ren JL,Lu YP,Wang L,Chen JM,Shi HX,Ye ZS,Wu YH,Zhong Y,Lin XT,Lin H,Pan JS,Luo JY.Express change of TFF1 in normal and injured gastric mucosa.Shijie Huaren Xiaohua Zazhi,2003,11(11):1809-1810.
[14]Li TL,Su YM,Qi F,Ni W,Zhang H,Zhang YC,Guo H.Effect of moxibustion on serum TFF and gastric mucosa ERK1/2 and PCNA in gastric ulcer model rats with spleen deficiency symptom.Hunan Zhongyiyao Daxue Xuebao,2015,35(2):49-51.
[15]Song J,Guo RF,Su RL,Qin EP,Wang JL.Expressions and significance of TFF2,CLDN18 and MUC5AC protein in gastric mucosal lesions.Wei Chang Bing Xue,2015,20(7):394-397.
[16]Zhang H,Guo H,Zhang YC,Liu M,Ai K,Su YM,Li MH,Li TL.Effect of moxibustion intervention on expression of gastric epidermal growth factor receptor and extracellular signal regulated kinase 1/2 expression in rats with gastric ulcer.Zhen Ci Yan Jiu,2014,39(5):351-357.
[17]Liu XL,Wang RJ,Zhao QQ.Effect on expressions of MUC5AC mRNA and protein in gastric mucosa of Bu Zhong Yi Qi decoction for rats with spleen deficiency.Zhongyao Yaoli Yu Linchuang,2012,28(4):7-9.
[18]Liu XL,Wang RJ,Zhang QQ.Effect on EGFR protein expression in gastric mucosa of Bu Zhong Yi Qi decoction for rats with spleen deficiency.Zhongyao Yaoli Yu Linchuang,2012,28(5):12-14.