健脾清肠方对DSS诱导结肠炎小鼠肠道动力学影响的研究
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摘要
[目的]探讨健脾清肠方(JQD)对DSS诱导结肠炎小鼠肠道动力的作用机制。[方法]C57BL/6小鼠随机分为Control组、DSS组、JQD组、5-ASA组。除Control组外,余各组小鼠用5%DSS自由饮用诱导结肠炎模型,造模7d后灌胃给药。第15天处死小鼠收集结肠组织,检测结肠组织病理学,ICC的超微结构,结肠组织IL-1β、IL-10、IFN-γ、TNF-α含量,c-kit mRNA、LC3-II mRNA、Beclin-l mRNA、NF-κB p65表达和结肠平滑肌张力。[结果]与DSS组比较,JQD组结肠黏膜表面上皮细胞移行修复,少量炎症细胞浸润,黏膜腺体基本恢复正常;ICC与其他细胞间连接完好,内部细胞器结构相对完整,可见到少量自噬泡存在;结肠组织IL-1β、TNF-α含量降低,IL-10、IFN-γ含量上升;c-kit mRNA表达上调,LC-3II mRNA、Beclin-1mRNA、NF-κB p65表达下降;结肠平滑肌条收缩振幅上升、收缩频率减少。[结论]JQD调节DSS模型小鼠异常肠道动力,其机制可能是通过抑制NF-κB/TNF-α通路及调节细胞因子IL-1β、IL-10、IFN-γ表达,抑制了肠道炎症的级联放大反应;抑制ICC过度自噬,调控ICC/SMC网络通路。
[Objective]This study was aimed to explore the mechanism underlying regulatory effect of Jianpi Qingchang decoction(JQD)on intestinal motility of DSS-induced colitis in mice.[Methods]C57BL/6mice were randomly divided into four groups:the control group,the DSS group,the JQD group,and the 5-ASA group.Except the control group,colitis was induced by giving distilled water containing 5%DSS.Seven days after modeling,the mice were administered corresponding drugs intragastrically.The mice were sacrificed on the 15 th day.The histopathologic lesions,and ultrastructure of colon ICC were observed.The levels of TNF-α,IL-1β,IL-10,IFN-γ,the expression of NF-κB p65,c-kit mRNA,LC3-II mRNA,Beclin-l mRNA,and the colonic smooth muscle tension were assessed.[Results]Compared with the DSS group,the colons presented the migration and repair of epithelial cells on the erosive mucosal surface with infiltration of fewer inflammatory cells and recovery of the glandular structure in the JQD group.The configuration of ICC was normal with intact connections between cells,and the ridge of mitochondria and a few autophagic vacuoles existed.The IL-1βand TNF-αlevels decreased;IL-10 and IFN-γlevels in-creased.C-kit mRNA expression increased;LC-3II mRNA,Beclin-1mRNA and NF-κB p65 expression decreased.The colonic smooth muscle contractile amplitude increased,and the contractile frequency decreased in the JQD group.[Conclusion]JQD can regulate the intestinal motility involved in inhibiting the NF-κB/TNF-αpathway,regulating IL-1β,IL-10,and IFN-γexpression,suppressing intestinal inflammatory cascade reaction,inhibiting excessive autophagy of ICC,and regulating the network path of ICC/SMC.
[Objective]This study was aimed to explore the mechanism underlying regulatory effect of Jianpi Qingchang decoction(JQD)on intestinal motility of DSS-induced colitis in mice.[Methods]C57BL/6mice were randomly divided into four groups:the control group,the DSS group,the JQD group,and the 5-ASA group.Except the control group,colitis was induced by giving distilled water containing 5%DSS.Seven days after modeling,the mice were administered corresponding drugs intragastrically.The mice were sacrificed on the 15 th day.The histopathologic lesions,and ultrastructure of colon ICC were observed.The levels of TNF-α,IL-1β,IL-10,IFN-γ,the expression of NF-κB p65,c-kit mRNA,LC3-II mRNA,Beclin-l mRNA,and the colonic smooth muscle tension were assessed.[Results]Compared with the DSS group,the colons presented the migration and repair of epithelial cells on the erosive mucosal surface with infiltration of fewer inflammatory cells and recovery of the glandular structure in the JQD group.The configuration of ICC was normal with intact connections between cells,and the ridge of mitochondria and a few autophagic vacuoles existed.The IL-1βand TNF-αlevels decreased;IL-10 and IFN-γlevels in-creased.C-kit mRNA expression increased;LC-3II mRNA,Beclin-1mRNA and NF-κB p65 expression decreased.The colonic smooth muscle contractile amplitude increased,and the contractile frequency decreased in the JQD group.[Conclusion]JQD can regulate the intestinal motility involved in inhibiting the NF-κB/TNF-αpathway,regulating IL-1β,IL-10,and IFN-γexpression,suppressing intestinal inflammatory cascade reaction,inhibiting excessive autophagy of ICC,and regulating the network path of ICC/SMC.
引文
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[9]Dai Y C,Tang Z P,Wang Z N,et al.Influence of shenqing recipe on morphology and quantity of colonic interstitial cells of cajal in trinitrobenzene sulfonic Acid induced rat colitis[J].Chin Med Sci J,2011,26(1):43-48.
[10]Rubinsztein D C,Codogno P,Levine B.Autophagy modulation as a potential therapeutic target for diverse diseases[J].Nat Rev Drug Discov,2012,11(9):709-730.
[11]戴彦成,张亚利,唐志鹏.ICC自噬调控——溃疡性结肠炎肠动力紊乱治疗的新靶点[J].胃肠病学,2015,20(6):377-379.
[12]Zhang D K,Cheng L N,Huang X L,et al.Tetrandrine ameliorates dextran-sulfate-sodium-induced colitis in mice through inhibition of nuclear factor-kappaB activation[J].Int J Colorectal Dis,2009,24(1):5-12.
[13]陈奇.中药药理研究方法学[M].北京:人民卫生出版社,2006:1103.
[14]Qureshi S,Song J,Lee H T,et al.CaM kinase II in colonic smooth muscle contributes to dysmotility in murine DSS-colitis[J].Neurogastroenterol Motil,2010,22(2):186-195.
[15]Liu X,Rusch N J,Striessnig J,et al.Down-regulation of L-type calcium channels in inflamed circular smooth muscle cells of the canine colon[J].Gastroenterology,2001,120(2):480-489.
[16]Ohama T,Hori M,Momotani E,et al.IL-1beta inhibits intestinal smooth muscle proliferation in an organ culture system:involvement of COX-2and iNOS induction in muscularis resident macrophages[J].Am J Physiol Gastrointest Liver Physiol,2007,292(5):G1315-1322.
[17]Rumessen J J.Ultrastructure of interstitial cells of Cajal at the colonic submuscular border in patients with ulcerative colitis[J].Gastroenterology,1996,111(6):1447-1455.
[18]Ledesma-soto Y,Callejas B E,Terrazas C A,et al.Extraintestinal Helminth Infection Limits Pathology and Proinflammatory Cytokine Expression during DSS-Induced Ulcerative Colitis:A Role for Alternatively Activated Macrophages and Prostaglandins[J].Biomed Res Int,2015,2015:563425.
[2]戴彦成,唐志鹏,李凯,等.溃疡性结肠炎中肠道动力学研究进展[J].世界华人消化杂志,2007,15(11):721-724.
[3]Bressler B,MarshalL J K,Bernstein C N,et al.Toronto Ulcerative Colitis Consensus Group.Clinical practice guidelines for the medical management of nonhospitalized ulcerative colitis:the Toronto consensus[J].Gastroenterology,2015,148(5):1035-1058.
[4]戴彦成,张亚利,唐志鹏.中医药治疗溃疡性结肠炎生存质量评价的现状与展望[J].世界中医药,2015,10(6):951-953.
[5]Dai Y C,Zhang Y L,Wang L J,et al.Clinical presentation and treatment strategies for ulcerative colitis:A retrospective study of 247inpatients[J].Chin J Integr Med,2016,22(11):811-816.
[6]戴彦成,郑烈,张亚利,等.健脾清肠方对溃疡性结肠炎患者生活质量的随机对照试验[J].中华中医药杂志,2015,31(5):1926-1932.
[7]张亚利,郭倩,戴彦成,等.健脾清肠方对脾虚湿热型激素依赖溃疡性结肠炎患RGα表达的影响[J].中国中西医结合消化杂志,2014,22(12):706-709.
[8]Negreanu L M,Assor P,Mateescu B,et al.Interstitial cells of Cajal in the gut--agastroenterologists point of view[J].World J Gastroenterol,2008,14(41):6285-6288.
[9]Dai Y C,Tang Z P,Wang Z N,et al.Influence of shenqing recipe on morphology and quantity of colonic interstitial cells of cajal in trinitrobenzene sulfonic Acid induced rat colitis[J].Chin Med Sci J,2011,26(1):43-48.
[10]Rubinsztein D C,Codogno P,Levine B.Autophagy modulation as a potential therapeutic target for diverse diseases[J].Nat Rev Drug Discov,2012,11(9):709-730.
[11]戴彦成,张亚利,唐志鹏.ICC自噬调控——溃疡性结肠炎肠动力紊乱治疗的新靶点[J].胃肠病学,2015,20(6):377-379.
[12]Zhang D K,Cheng L N,Huang X L,et al.Tetrandrine ameliorates dextran-sulfate-sodium-induced colitis in mice through inhibition of nuclear factor-kappaB activation[J].Int J Colorectal Dis,2009,24(1):5-12.
[13]陈奇.中药药理研究方法学[M].北京:人民卫生出版社,2006:1103.
[14]Qureshi S,Song J,Lee H T,et al.CaM kinase II in colonic smooth muscle contributes to dysmotility in murine DSS-colitis[J].Neurogastroenterol Motil,2010,22(2):186-195.
[15]Liu X,Rusch N J,Striessnig J,et al.Down-regulation of L-type calcium channels in inflamed circular smooth muscle cells of the canine colon[J].Gastroenterology,2001,120(2):480-489.
[16]Ohama T,Hori M,Momotani E,et al.IL-1beta inhibits intestinal smooth muscle proliferation in an organ culture system:involvement of COX-2and iNOS induction in muscularis resident macrophages[J].Am J Physiol Gastrointest Liver Physiol,2007,292(5):G1315-1322.
[17]Rumessen J J.Ultrastructure of interstitial cells of Cajal at the colonic submuscular border in patients with ulcerative colitis[J].Gastroenterology,1996,111(6):1447-1455.
[18]Ledesma-soto Y,Callejas B E,Terrazas C A,et al.Extraintestinal Helminth Infection Limits Pathology and Proinflammatory Cytokine Expression during DSS-Induced Ulcerative Colitis:A Role for Alternatively Activated Macrophages and Prostaglandins[J].Biomed Res Int,2015,2015:563425.