茯苓甘草汤对功能性消化不良模型大鼠的改善作用机制研究
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摘要
目的:研究茯苓甘草汤对功能性消化不良(FD)模型大鼠的改善作用机制。方法:取SD大鼠60只,随机分为正常组、模型组、茯苓甘草汤高剂量组(20 g/kg)、茯苓甘草汤低剂量组(10 g/kg)、联用组(茯苓甘草汤10 g/kg+多潘立酮0.004 g/kg)、多潘立酮组(0.004 g/kg),每组10只。除正常组外,其余组大鼠均灌胃冰稀盐酸建立FD模型。造模结束后,正常组和模型组大鼠灌胃等体积常温蒸馏水,其余组大鼠灌胃相应常温药物溶液(1 mL/100 g),每天1次,连续7 d。测定大鼠胃内残渣量以评价胃排空功能。采用免疫组化SP法检测胃黏膜组织中水通道蛋白3(AQP3)、胃促生长素(Ghrelin)、P物质(Substance P)的蛋白表达水平;采用蛋白印迹法检测胃黏膜组织中紧密连接蛋白1(Claudin-1)、闭锁蛋白(Occludin)、血管活性肠肽(VIP)的蛋白表达水平。结果:与正常组比较,模型组大鼠胃内残渣量明显增多(P<0.01);胃黏膜组织中AQP3、Ghrelin蛋白阳性表达明显减少,Substance P蛋白阳性表达较多;AQP3、Ghrelin、Claudin-1、Occludin蛋白表达水平显著降低,Substance P、VIP蛋白表达水平显著升高(P<0.05或P<0.01)。与模型组比较,各给药组大鼠胃内残渣量明显减少(P<0.05或P<0.01);各给药组大鼠胃黏膜组织中AQP3、Ghrelin蛋白阳性表达较多,Substance P蛋白阳性表达较少;各给药组大鼠胃黏膜组织中AQP3、Occludin蛋白表达水平显著升高,VIP蛋白表达水平显著降低(P<0.05或P<0.01);茯苓甘草汤高剂量组、茯苓甘草汤低剂量组和联用组大鼠胃黏膜组织中Ghrelin、Claudin-1蛋白表达水平显著升高,Substance P蛋白表达水平显著降低(P<0.05)。结论:茯苓甘草汤可能通过对大鼠胃黏膜组织中AQP3、Ghrelin、Claudin-1、Occludin蛋白表达的上调和Substance P、VIP蛋白表达的下调,从而改善FD模型大鼠水液潴留症状。
OBJECTIVE:To study improvement effects of Fuling gancao decoction on functional dyspepsia (FD)model rats.METHODS:Sixty SD rats were randomly divided into normal group,model group,Fuling gancao decoction high-dose group (20 g/kg), Fuling gancao decoction low-dose group (10 g/kg), drug combination group (Fuling gancao decoction 10 g/kg +domperidone 0.004 g/kg),domperidone group (0.004 g/kg),with 10 rats in each group. Except for normal group,other groupswere givenicy dilute hydrochloric acid intragastrically to establish FD model. After modeling,normal group and model group weregiven constant volume of distilled water at room temperature intragastrically,and other groups were given relevant drug solution (1 mL/100 g) intragastrically,once a day,for consecutive 7 d. The amount of gastric residue was measured to evaluate the gastricemptying function. Immunohistochemistry SP method was used to detect the protein expression of AQP3,Ghrelin and Substance Pin gastric mucosa tissue. The protein expression of Claudin-1,Occludin and VIP were detected by Western blot assay. RESULTS:Compared with normal group,the amount of gastric residue was increased significantly in model group (P<0.01);positiveexpression of AQP3 and Ghrelin protein in gastric mucosa tissue was decreased significantly,while the positive expression ofSubstance P protein tended to increase;the protein expression levels of AQP3,Ghrelin,Claudin-1 and Occludin were decreasedsignificantly,while those of Substance P and VIP were increased significantly (P<0.05 or P<0.01). Compared with model group,the amount of gastric residue was decreased significantly in administration groups (P<0.05 or P<0.01);the positive expression ofAQP3 and Ghrelin protein tended to increase,while less positive expression of Substance P was found;the protein expressionlevels of AQP3 and Occludin in gastric mucosa tissue were increased significantly in administration groups,while those of VIPwere decreased significantly (P<0.05 or P<0.01);the protein expression levels of Ghrelin and Claudin-1 in gastric mucosa tissuewere increased significantly in Fuling gancao decoction high-dose group,Fuling gancao decoction low-dose group and drugcombination group,while those of Substance P were decreased significantly (P<0.05). CONCLUSIONS:Fuling gancao decoctionmay improve the symptom of fluid retention in FD model rats by up-regulating the protein expression of AQP3,Ghrelin,Claudin-1 and Occludin and down-regulating the protein expression of Substance P and VIP in gastric mucosa tissue of rats.
OBJECTIVE:To study improvement effects of Fuling gancao decoction on functional dyspepsia (FD)model rats.METHODS:Sixty SD rats were randomly divided into normal group,model group,Fuling gancao decoction high-dose group (20 g/kg), Fuling gancao decoction low-dose group (10 g/kg), drug combination group (Fuling gancao decoction 10 g/kg +domperidone 0.004 g/kg),domperidone group (0.004 g/kg),with 10 rats in each group. Except for normal group,other groupswere givenicy dilute hydrochloric acid intragastrically to establish FD model. After modeling,normal group and model group weregiven constant volume of distilled water at room temperature intragastrically,and other groups were given relevant drug solution (1 mL/100 g) intragastrically,once a day,for consecutive 7 d. The amount of gastric residue was measured to evaluate the gastricemptying function. Immunohistochemistry SP method was used to detect the protein expression of AQP3,Ghrelin and Substance Pin gastric mucosa tissue. The protein expression of Claudin-1,Occludin and VIP were detected by Western blot assay. RESULTS:Compared with normal group,the amount of gastric residue was increased significantly in model group (P<0.01);positiveexpression of AQP3 and Ghrelin protein in gastric mucosa tissue was decreased significantly,while the positive expression ofSubstance P protein tended to increase;the protein expression levels of AQP3,Ghrelin,Claudin-1 and Occludin were decreasedsignificantly,while those of Substance P and VIP were increased significantly (P<0.05 or P<0.01). Compared with model group,the amount of gastric residue was decreased significantly in administration groups (P<0.05 or P<0.01);the positive expression ofAQP3 and Ghrelin protein tended to increase,while less positive expression of Substance P was found;the protein expressionlevels of AQP3 and Occludin in gastric mucosa tissue were increased significantly in administration groups,while those of VIPwere decreased significantly (P<0.05 or P<0.01);the protein expression levels of Ghrelin and Claudin-1 in gastric mucosa tissuewere increased significantly in Fuling gancao decoction high-dose group,Fuling gancao decoction low-dose group and drugcombination group,while those of Substance P were decreased significantly (P<0.05). CONCLUSIONS:Fuling gancao decoctionmay improve the symptom of fluid retention in FD model rats by up-regulating the protein expression of AQP3,Ghrelin,Claudin-1 and Occludin and down-regulating the protein expression of Substance P and VIP in gastric mucosa tissue of rats.
引文
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[12]孙杰,桑凯,王东晓,等.肺脾两虚型COPD模型大鼠下丘脑、胃组织Ghrelin及其受体的表达变化[J].中国实验方剂学杂志,2018,24(4):124-130.
[13]叶贝,黄永坤.P-物质、血管活性肠肽、胃泌素和5-羟色胺与胃肠功能的研究进展[J].国际儿科学杂志,2017,44(4):249-252.
[14]骆凤,孙国瑛,贺丽萍,等.血管活性肠肽与各系统疾病内在关联的研究进展[J].解剖学杂志,2016,39(5):626-628.
[15]张欢,付立波,周慧,等.P物质在痛觉调节作用的研究进展[J].长春师范大学学报,2018,37(2):180-182.
[16]BARRETO TR,COSTOLA-DE-SOUZA C,MARGATHORO,et al.Repeated domperidone treatment modulates pulmonary cytokines in LPS-induced acute lung injury in mice[J].Int Immunopharmacol,2018,56(6):43-55.
[2]曹峰,傅延龄.茯苓甘草汤对功能性消化不良大鼠胃底NO、ACHE影响的实验研究[J].贵阳中医学院学报,2011,33(4):43-46.
[3]ZHANG G,XIE S,HU W,et al.Effects of electroacupuncture on interstitial cells of cajal(ICC)ultrastructure and connexin 43 protein expression in the gastrointestinal tract of functional dyspepsia(FD)rats[J].Med Sci Monit,2016.DOI:10.12659/MSM.899023.
[4]LI XL,ZHANG SS,YANG C,et al.Effect of Zhizhu pill on gastric smooth muscle contractile response and protein expression of growth hormone secretagogue receptor in functional dyspepsia rats[J].Zhongguo Zhong Xi Yi Jie He Za Zhi,2016,36(2):210-215.
[5]程寒,王垂杰.中西医治疗功能性消化不良研究进展[J].辽宁中医药大学学报,2017,19(10):219-221.
[6]杨文广,刘露路,王珏,等.慢性炎症与功能性消化不良研究进展[J].现代医药卫生,2017,33(12):1801-1803.
[7]徐雯,王楠,丁浩然,等.广藿香对湿阻中焦证大鼠胃肠功能的影响[J].中国中药杂志,2017,42(23):4649-4655.
[8]LI Z,LI B,ZHANG L,et al.The proliferation impairment induced by AQP3 deficiency is the result of glycerol uptake and metabolism inhibition in gastric cancer cells[J].Tumour Biol,2016,37(7):9169-9179.
[9]邢晓辉,李力仙,郭天林,等.Occludin蛋白与细胞间紧密连接关系及临床意义[J].现代生物医学进展,2015,15(8):1553-1555.
[10]CUNNIFFE C,BRANKIN B,LAMBKIN H,et al.The role of Claudin-1 and Claudin-7 in cervical tumorigenesis[J].Anticancer Res,2014,34(6):2851-2857.
[11]刘洋,钟逸凡,苏曼卿,等.顺铂处理大鼠Ghrelin的变化及意义[J].青岛科技大学学报,2017,38(5):40-43.
[12]孙杰,桑凯,王东晓,等.肺脾两虚型COPD模型大鼠下丘脑、胃组织Ghrelin及其受体的表达变化[J].中国实验方剂学杂志,2018,24(4):124-130.
[13]叶贝,黄永坤.P-物质、血管活性肠肽、胃泌素和5-羟色胺与胃肠功能的研究进展[J].国际儿科学杂志,2017,44(4):249-252.
[14]骆凤,孙国瑛,贺丽萍,等.血管活性肠肽与各系统疾病内在关联的研究进展[J].解剖学杂志,2016,39(5):626-628.
[15]张欢,付立波,周慧,等.P物质在痛觉调节作用的研究进展[J].长春师范大学学报,2018,37(2):180-182.
[16]BARRETO TR,COSTOLA-DE-SOUZA C,MARGATHORO,et al.Repeated domperidone treatment modulates pulmonary cytokines in LPS-induced acute lung injury in mice[J].Int Immunopharmacol,2018,56(6):43-55.
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