加味四君子汤对脾虚犬小肠组织形态及EGF表达的影响
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摘要
【目的】探究加味四君子汤对脾虚犬小肠组织形态及EGF表达的影响。【方法】选择1岁龄的比格犬24只,随机分为4组,每组6只,分别为对照组、脾虚组、自然恢复组、中药恢复组。对照组犬饲喂基础饲粮;脾虚组、自然恢复组、中药恢复组犬每天以6 mL/kg番泻叶灌喂造模。第7天,将脾虚组犬解剖采样,中药恢复组在基础饲粮中添加2%加味四君子汤,自然恢复组饲喂基础饲粮。第14天,全部犬解剖采样。采集犬十二指肠、空肠中段、回肠进行HE染色,测量肠绒毛长度、隐窝深度,计算V/C值;检测小肠EGF蛋白表达量以及mRNA相对表达量。【结果】与对照组相比,脾虚组犬小肠出现弥漫性的肠道黏膜充血、水肿,并有大量黏液渗出;小肠绒毛稀疏、变短;肠绒毛长度极显著降低(P<0.01),隐窝深度极显著升高(P<0.01),十二指肠、回肠V/C值显著降低(P<0.05),空肠V/C值极显著降低(P<0.01);小肠EGF的蛋白表达量极显著降低(P<0.01),mRNA表达量显著降低(P<0.05)。用加味四君子汤治疗后,中药恢复组各项指标均恢复到接近对照组水平。【结论】加味四君子汤能够使番泻叶诱导犬脾虚证的小肠组织形态、EGF蛋白表达量以及mRNA相对表达量恢复正常,对犬脾虚证有明显的治疗效果。
[Purpose]The present study was conducted to investigate the effects of supplementary Sijunzi decoction on the tissue morphology and EGF expression of small intestine in splenic asthenia canines.[Method]Twenty-four one-year-old beagles were selected and randomly divided into four groups(6 canines in each group). The four groups were control group, splenic asthenia group, natural recovery group and Chinese medicine recovery group, respectively. The canines in control group were fed a basal diet. The canines in splenic asthenia group, natural recovery group and Chinese medicine recovery group were fed with Senna at 6 mL/kg a day. On the 7 th day, the canines in splenic asthenia group were dissected for collecting samples, the canines in Chinese medicine recovery group were fed a basal diet with 2% supplementary Sijunzi decoction, and in natural recovery group were fed a basal diet. On the 14 th day, all canines were dissected for collecting samples. Duodenum, jejunum and ileum of canines were collected to measure villus length, crypt depth and V/C values after HE staining and to detect EGF protein expression and EGF mRNA relative expression levels in small intestine.[Result]Compared with the canines in control group, the diffuse congestion and edema appeared in the small intestinal mucosa of canines in splenic asthenia group, and a large amount of mucous oozed out. The villi were sparser and shorter in small intestines, villus length extremely significantly decreased(P<0.01), crypt depth extremely significantly increased(P<0.01), V/C of duodenum and ileum significantly decreased(P<0.05) and V/C of jejunum extremely significantly decreased(P<0.01). EGF protein expression in small intestine significantly decreased(P<0.05) and EGF mRNA relative expression levels in small intestine extremely significantly decreased(P<0.01). After treating by supplementary Sijunzi decoction, the above indexes of the canines in Chinese medicine recovery group recovered close to the control group.[Conclusion]Supplementary Sijunzi decoction can improve the tissue morphology, EGF protein expression and EGF mRNA relative expression levels in small intestine, and has obvious effects on treating the canines with splenic asthenia syndrome.
[Purpose]The present study was conducted to investigate the effects of supplementary Sijunzi decoction on the tissue morphology and EGF expression of small intestine in splenic asthenia canines.[Method]Twenty-four one-year-old beagles were selected and randomly divided into four groups(6 canines in each group). The four groups were control group, splenic asthenia group, natural recovery group and Chinese medicine recovery group, respectively. The canines in control group were fed a basal diet. The canines in splenic asthenia group, natural recovery group and Chinese medicine recovery group were fed with Senna at 6 mL/kg a day. On the 7 th day, the canines in splenic asthenia group were dissected for collecting samples, the canines in Chinese medicine recovery group were fed a basal diet with 2% supplementary Sijunzi decoction, and in natural recovery group were fed a basal diet. On the 14 th day, all canines were dissected for collecting samples. Duodenum, jejunum and ileum of canines were collected to measure villus length, crypt depth and V/C values after HE staining and to detect EGF protein expression and EGF mRNA relative expression levels in small intestine.[Result]Compared with the canines in control group, the diffuse congestion and edema appeared in the small intestinal mucosa of canines in splenic asthenia group, and a large amount of mucous oozed out. The villi were sparser and shorter in small intestines, villus length extremely significantly decreased(P<0.01), crypt depth extremely significantly increased(P<0.01), V/C of duodenum and ileum significantly decreased(P<0.05) and V/C of jejunum extremely significantly decreased(P<0.01). EGF protein expression in small intestine significantly decreased(P<0.05) and EGF mRNA relative expression levels in small intestine extremely significantly decreased(P<0.01). After treating by supplementary Sijunzi decoction, the above indexes of the canines in Chinese medicine recovery group recovered close to the control group.[Conclusion]Supplementary Sijunzi decoction can improve the tissue morphology, EGF protein expression and EGF mRNA relative expression levels in small intestine, and has obvious effects on treating the canines with splenic asthenia syndrome.
引文
[1]刘大鹏.脾虚在功能性胃肠病中辨证意义及相关规律的研究[D].济南:山东中医药大学,2014.
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[8]刘静.犬脾虚模型的建立及中药制剂的治疗效果[D].合肥:安徽农业大学,2016.
[9]肖新云,邓艳玲,刘又嘉,等.番泻叶所致脾虚泄泻小鼠血常规的研究[J].湖北中医药大学学报,2016,18(6):49.DOI:10.3969/j.issn.1008-987x.2016.06.15.
[10]石达友,李盛强,朱华君,等.四君子汤对脾虚犬体重及部分血清生化指标的影响[J].动物医学进展,2010,31(9):29.DOI:10.3969/j.issn.1007-5038.2010.09.007.
[11]王月婷.不同频率摩腹对脾虚家兔胃肠黏膜形态影响的研究[D].广州:广州中医药大学,2017.
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[13]陈功义,郝振芳,肖传斌.大豆异黄酮对固始鸡小肠形态结构的影响[J].动物营养学报,2014,26(11):3426.DOI:10.3969/j.issn.1006-267x.2014.11.032.
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[16]李茹柳,陶玉珠,温鹏,等.甘草黄酮提取物对小肠上皮细胞IEC-6迁移及多胺的影响[J].中草药,2013,44(19):2722.DOI:10.7501/j.issn.0253-2670.2013.19.017.
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[19]汤小朋,刘虎,杨淑芬,等.表皮生长因子在动物肠道无机离子及其他营养物质吸收中的作用[J].动物营养学报,2016,28(8):2317.DOI:10.3969/j.issn.1006-267x.2016.08.001.
[20]汤小朋,方热军.表皮生长因子对早期断奶仔猪生长性能及肠道健康的影响[J].动物营养学报,2015,27(11):3345.DOI:10.3969/j.issn.1006-267x.2015.11.005.
[21]TANG X P,LIU H,YANG S F,et al.Epidermal growth factor and intestinal barrier function[J].Mediators of Inflammation,2016:1927348.DOI:10.1155/2016/1927348.
[22]ABUD H,WATSON N J.Growth of intestinal epithelium in organ culture is dependent on EGF signalling[J].Experimental Cell Research,2005,303(2):252.DOI:10.1016/j.yexcr.2004.10.006.
[23]范志勇,王康宁,周定刚.不同水平表皮生长因子对早期断奶仔猪生产性能、血清理化指标及免疫功能的影响[J].动物营养学报,2005,17(3):41.
[24]任丹丹,贺胜男,曹程鸣,等.加味四君子汤对仔猪消化吸收功能及小肠EGF表达的影响[J].中国兽医科学,2016(8):1065.
[2]吕林,唐旭东,王凤云,等.从“脾主运化”理论探讨论治功能性胃肠病[J].时珍国医国药,2016(1):160.
[3]纪云飞,王瑞君,李晓波.复方四君子汤的化学成分和药理作用研究进展[J].中草药,2016,47(5):837.
[4]吕苑.四君子汤的药理研究和临床应用[J].中医研究,2012,25(1):76.DOI:10.3969/j.issn.1001-6910.2012.01.037.
[5]刘双,杨静,江振作,等.中药“神曲”发酵工艺及质量标准研究进展[J].天津中医药,2015,32(5):318.
[6]JURIKOVA T,SOCHOR J,ROP O,et al.Polyphenolic profile and biological activity of Chinese hawthorn(Crataegus pinnatifida BUNGE)fruits[J].Molecules,2012,17(12):14490.DOI:10.3390/molecules171214490.
[7]国家药典委员会.中华人民共和国药典:一部[M].北京:化学工业出版社,2005.
[8]刘静.犬脾虚模型的建立及中药制剂的治疗效果[D].合肥:安徽农业大学,2016.
[9]肖新云,邓艳玲,刘又嘉,等.番泻叶所致脾虚泄泻小鼠血常规的研究[J].湖北中医药大学学报,2016,18(6):49.DOI:10.3969/j.issn.1008-987x.2016.06.15.
[10]石达友,李盛强,朱华君,等.四君子汤对脾虚犬体重及部分血清生化指标的影响[J].动物医学进展,2010,31(9):29.DOI:10.3969/j.issn.1007-5038.2010.09.007.
[11]王月婷.不同频率摩腹对脾虚家兔胃肠黏膜形态影响的研究[D].广州:广州中医药大学,2017.
[12]RIEGER J,JANCZYK P,HüNIGEN H,et al.Intraepithelial lymphocyte numbers and histomorphological parameters in the porcine gut after Enterococcus faecium NCIMB 10415 feeding in a Salmonella Typhimurium challenge[J].Veterinary Immunology and Immunopathology,2015,164(1/2):40.DOI:10.1016/j.vetimm.2014.12.013.
[13]陈功义,郝振芳,肖传斌.大豆异黄酮对固始鸡小肠形态结构的影响[J].动物营养学报,2014,26(11):3426.DOI:10.3969/j.issn.1006-267x.2014.11.032.
[14]YE L,GONG J W,WANG Y H,et al.Pharmacological activities of Sijunzi decoction which are related to its antioxidant properties[J].Journal of Chemistry,2014(33):1.DOI:10.1155/2014/278318.
[15]YU X Y,CUI Z G,ZHOU Z L,et al.Si-Jun-Zi decoction treatment promotes the restoration of intestinal function after obstruction by regulating intestinal homeostasis[J].Evidence-Based Complementary and Alternative Medicine,2014(11):928579.DOI:10.1155/2014/928579.
[16]李茹柳,陶玉珠,温鹏,等.甘草黄酮提取物对小肠上皮细胞IEC-6迁移及多胺的影响[J].中草药,2013,44(19):2722.DOI:10.7501/j.issn.0253-2670.2013.19.017.
[17]BARNARD J A,BEAUCHAMP R D,RUSSELL W E,et al.Epidermal growth factor-related peptides and their relevance to gastrointestinal pathophysiology[J].Gastroenterology,1995,108(2):564.DOI:10.1016/0016-5085(95)90087-X.
[18]PLAYFORD R J,WRIGHT N A.Why is epidermal growth factor present in the gut lumen?[J].Gut,1996,38(3):303.DOI:10.1136/gut.38.3.303.
[19]汤小朋,刘虎,杨淑芬,等.表皮生长因子在动物肠道无机离子及其他营养物质吸收中的作用[J].动物营养学报,2016,28(8):2317.DOI:10.3969/j.issn.1006-267x.2016.08.001.
[20]汤小朋,方热军.表皮生长因子对早期断奶仔猪生长性能及肠道健康的影响[J].动物营养学报,2015,27(11):3345.DOI:10.3969/j.issn.1006-267x.2015.11.005.
[21]TANG X P,LIU H,YANG S F,et al.Epidermal growth factor and intestinal barrier function[J].Mediators of Inflammation,2016:1927348.DOI:10.1155/2016/1927348.
[22]ABUD H,WATSON N J.Growth of intestinal epithelium in organ culture is dependent on EGF signalling[J].Experimental Cell Research,2005,303(2):252.DOI:10.1016/j.yexcr.2004.10.006.
[23]范志勇,王康宁,周定刚.不同水平表皮生长因子对早期断奶仔猪生产性能、血清理化指标及免疫功能的影响[J].动物营养学报,2005,17(3):41.
[24]任丹丹,贺胜男,曹程鸣,等.加味四君子汤对仔猪消化吸收功能及小肠EGF表达的影响[J].中国兽医科学,2016(8):1065.