电针足三里调控SCF/c-Kit信号通路对功能性消化不良大鼠胃窦组织肌内Cajal间质细胞的作用
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摘要
目的观察电针足三里介导SCF/c-Kit信号通路对功能性消化不良大鼠胃窦组织肌内Cajal间质细胞的影响,探讨电针足三里治疗功能性消化不良的作用机制。方法将40只SD大鼠随机分别为空白组、模型组、电针组、假电针组和电针加阻断剂组。除空白组外,其余各组均采用郭氏夹尾+不规则喂养+冰生理盐水灌胃法的多因素干预造模。在造模第5天,给予电针加阻断剂组腹腔注射Kit单克隆抗体(ACK2)。造模14d后,电针组取双侧足三里治疗,假电针组只针刺不接通电流进行治疗。10d治疗结束后,用免疫印迹法检测胃窦组织的c-Kit与SCF蛋白的表达量;用RT-qPCR测量各组大鼠胃窦组织的c-Kit的m RNA表达水平;在透射电子显微镜下观察各组大鼠胃窦组织中的肌内Cajal间质细胞的超微结构变化。结果与空白组对比,模型组c-Kit、SCF蛋白的表达量和c-Kit m RNA表达水平显著下降(<0.01);与模型组对比,电针组胃窦组织中c-Kit、SCF蛋白的表达量和c-Kit m RNA表达水平增加(<0.05),电针加阻断剂组c-Kit蛋白含量与c-Kit m RNA表达降低,SCF蛋白含量出现增高趋势,差异均无统计学意义(均>0.05);与电针组对比,假电针组SCF、c-Kit蛋白含量与c-Kit m RNA表达水平降低(<0.05)。在透射电镜下,电针组肌内Cajal间质细胞较模型组、假电针组、电针加阻断剂组数量增多,与平滑肌和神经末梢的连接更密切。结论电针足三里可以增加Cajal间质细胞的数量和修复受损的网状结构,从而改善Cajal间质细胞网状结构功能。这种作用机制可能是通过调节SCF、c-Kit蛋白含量与c-Kit m RNA表达,从而调控SCF/c-Kit信号通路实现的。
引文
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[24] Powley TL,Phillips RJ. Vagal intramuscular array afferents form complexes with interstitial cells of Cajal in gastrointestinal smooth muscle:analogues of muscle spindle organs[J].Neuroscience,2011(186):188-200.
[25] Gomez Pinilla PJ,Gibbons SJ,Sarr MG,et al. Changes in interstitial cells of cajal with age in the human stomach and colon[J]. Neurogastroenterology&Motility,2011,23(1):36-44.
[26]朱金照. Cajal间质细胞在胃肠动力疾病中的作用[J].医学研究生学报,2004,17(6):562-564.
[27] Torihashi S,Nishi K,Tokutomi Y,et al. Blockade of kit signaling induces transdifferentiation of interstitial cells of Cajal to a smooth muscle phenotype[J]. Gastroenterology,1999,117(1):140-148.
[28] Takeda M,Takayama I,Terada N,et al. Immunoelectronmicroscopic study of Kit-expressing cells in the jejunum of wildtype and Ws/Ws rats[J]. Cell and tissue research,2001,304(1):21-30.
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[30]杨敏,张红星,邹燃.针刺对功能性消化不良症状及胃动力的影响[J].中国康复,2009,24(2):100-102.
[31] Huizinga JD,Zarate N,Farrugia G. Physiology,injury,and recovery of interstitial cells of Cajal:basic and clinical science[J]. Gastroenterology,2009,137(5):1548-1556.
[2] Ghoshal UC,Singh R,Chang FY,et al. Epidemiology of uninvestigated and functional dyspepsia in Asia:facts and fiction[J]. Journal of neurogastroenterology and motility,2011,17(3):235.
[3] Brook RA,Kleinman NL,Melkonian AK,et al. Functional dyspepsia impacts absenteeism and direct and indirect costs[J]. Clinical Gastroenterology and Hepatology,2010,8(6):498-503.
[4] Min YW. The Implication of Gastric Dysmotility in the Pathophysiology of Functional Dyspepsia[J]. Journal of neurogastroenterology and motility,2017,23(3):323.
[5] Klein S,Seidler B,Kettenberger A,et al. Interstitial cells of Cajal integrate excitatory and inhibitory neurotransmission with intestinal slow-wave activity[J]. Nature communications,2013(4):1630.
[6] Ward SM,Mclaren GJ,Sanders KM. Interstitial cells of Cajal in the deep muscular plexus mediate enteric motor neurotransmission in the mouse small intestine[J]. Journal of Physiology,2010,573(1):147-159.
[7] Tong W,Jia H,Zhang L,et al. Exogenous stem cell factor improves interstitial cells of Cajal restoration after blockade of c-kit signaling pathway[J]. Scandinavian journal of gastroenterology,2010,45(7-8):844-851.
[8]郭海军,林洁,李国成,等.功能性消化不良的动物模型研究[J].中国中西医结合消化杂志,2001,9(3):141-142.
[9]王煜姣,凌江红,张钰琴,等.复合病因造模法制备功能性消化不良大鼠模型[J].世界华人消化杂志,2014(2):210-214.
[10]徐派的,张红星,杨云,等.电针对功能性消化不良模型大鼠胃肠动力及VIP和CGRP水平的影响[J].中国中西医结合杂志,2017(3):360-364.
[11]沈波.趋化因子受体5在大鼠心肌缺血再灌注损伤中的作用及机制研究[D].武汉:武汉大学,2013.
[12]李思宇.针刺足三里对功能性消化不良小鼠脑肠肽SCF/c-kit表达影响的研究[D].成都:成都中医药大学,2016.
[13]郭义,方剑乔.实验针灸学[M].北京:中国中医药出版社,2012.
[14] Carbone F,Tack J. Gastroduodenal mechanisms underlying functional gastric disorders[J]. Digestive Diseases,2014,32(3):222-229.
[15] Samsom M,Verhagen MA,vanBerge Henegouwen GP,et al. Abnormal clearance of exogenous acid and increased acid sensitivity of the proximal duodenum in dyspeptic patients[J]. Gastroenterology,1999,116(3):515-520.
[16] FarréR,Vanheel H,Vanuytsel T,et al. In functional dyspepsia,hypersensitivity to postprandial distention correlates with meal-related symptom severity[J]. Gastroenterology,2013,145(3):566-573.
[17] Asano H,Tomita T,Nakamura K,et al. Prevalence of gastric motility disorders in patients with functional dyspepsia[J]. Journal of neurogastroenterology and motility,2017,23(3):392.
[18] Langton P,Ward SM,Carl A,et al. Spontaneous electrical activity of interstitial cells of Cajal isolated from canine proximal colon[J]. Proceedings of the National Academy of Sciences,1989,86(18):7280-7284.
[19] Torihashi S,Ward SM,Nishikawa SI,et al. c-kit-Dependent development of interstitial cells and electrical activity in the murine gastrointestinal tract[J]. Cell and tissue research,1995,280(1):97-111.
[20] Vanderwinden JM,Rumessen JJ. Interstitial cells of Cajal in human gut and gastrointestinal disease[J]. Microscopy research and technique,1999,47(5):344-360.
[21] Wang XY,Sanders KM,Ward SM. Intimate relationship between interstitial cells of Cajal and enteric nerves in the guinea-pig small intestine[J]. Cell and tissue research,1999,295(2):247-256.
[22] Toma H,Nakamura K,Emson PC,et al. Immunohistochemical distribution of c-Kit-positive cells and nitric oxide synthase-positive nerves in the guinea-pig small intestine[J].Journal of the autonomic nervous system,1999,75(2):93-99.
[23] Hashitani H,Garcia-Londo?o AP,Hirst GDS,et al. Atypical slow waves generated in gastric corpus provide dominant pacemaker activity in guinea pig stomach[J]. J Physiol,2010,569(2):459-465.
[24] Powley TL,Phillips RJ. Vagal intramuscular array afferents form complexes with interstitial cells of Cajal in gastrointestinal smooth muscle:analogues of muscle spindle organs[J].Neuroscience,2011(186):188-200.
[25] Gomez Pinilla PJ,Gibbons SJ,Sarr MG,et al. Changes in interstitial cells of cajal with age in the human stomach and colon[J]. Neurogastroenterology&Motility,2011,23(1):36-44.
[26]朱金照. Cajal间质细胞在胃肠动力疾病中的作用[J].医学研究生学报,2004,17(6):562-564.
[27] Torihashi S,Nishi K,Tokutomi Y,et al. Blockade of kit signaling induces transdifferentiation of interstitial cells of Cajal to a smooth muscle phenotype[J]. Gastroenterology,1999,117(1):140-148.
[28] Takeda M,Takayama I,Terada N,et al. Immunoelectronmicroscopic study of Kit-expressing cells in the jejunum of wildtype and Ws/Ws rats[J]. Cell and tissue research,2001,304(1):21-30.
[29]周利,程艳萍.电针对功能性消化不良大鼠的胃排空和血浆Ghrelin的调节作用[J].湖北中医杂志,2014(3):19-21.
[30]杨敏,张红星,邹燃.针刺对功能性消化不良症状及胃动力的影响[J].中国康复,2009,24(2):100-102.
[31] Huizinga JD,Zarate N,Farrugia G. Physiology,injury,and recovery of interstitial cells of Cajal:basic and clinical science[J]. Gastroenterology,2009,137(5):1548-1556.