3种石斛对脾阴虚型大鼠便秘的治疗作用及其机制初步研究
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摘要
本草记载石斛具有滋阴强阴的功效,临床用于脾阴虚引起便秘。霍山石斛作为石斛众多品种中独立的物种,还未有脾阴虚便秘的实验研究报道。该实验旨在说明霍山石斛对脾阴虚型大鼠便秘的治疗作用及其初步机制,并与《中国药典》中收载的金钗石斛和铁皮石斛进行比较,明确其作用特点。采用复合因素法对70只大鼠复制脾阴虚模型,模型大鼠随机分为7组:模型组(Model),六味地黄丸组(LWDHP),霍山石斛高(DHS-H)、中(DHS-M)、低(DHS-L)剂量组以及铁皮石斛组(DNS),金钗石斛组(DOS),另选10只作为正常组(Normal)。连续给药7 d后,检测各组大鼠的粪便含水率和小肠推进率; HE染色观察各组大鼠回肠、结肠的病理损伤情况;免疫组化法和Western blot法检测水通道蛋白3(AQP3)的表达水平;酶联免疫法检测各组大鼠回肠组织中的生长抑素(SS)和胃动素(MTL)。结果显示,与模型组相比,各给药组大鼠12 h内粪便粒数增多、粪便含水率升高,且小肠推进率升高(P<0. 01),回肠和结肠的病理损伤明显减轻,AQP3蛋白表达明显降低(P<0. 01),且能明显增加大鼠回肠组织中胃动素(MTL)水平、降低生长抑素(SS)水平(P<0. 01),霍山石斛各剂量组显示出良好的量效关系。因此,霍山石斛对大鼠脾阴虚型便秘模型具有显著的治疗作用,其作用机制可能与肠动力和水液代谢有关。
Books on Chinese herbal medicines have shown that Dendrobium has the effect of nourishing Yin and reinforcing Yin,usually used for constipation induced by spleen Yin deficiency in clinical application. D. huoshanense,as an independent species among many species of Dendrobium,has no experimental studies about its effects on spleen Yin deficiency-type constipation. The purpose of this experiment was to illustrate the therapeutic effect of D. huoshanense on the constipation of spleen Yin deficiency type in rats,investigate its preliminary mechanism,and compare it with the D. officinale and D. nobile contained in the Chinese Pharmacopoeia to clarify its characteristics. The spleen Yin deficiency model was replicated in 70 rats by the composite factor method,and then the model rats were randomly divided into 7 groups: model group,Liuwei Dihuang Pills group( LWDHP),D. huoshanense high( DHS-H),medium( DHS-M),low( DHS-L) dose groups,D. nobile group( DNS),and D. officinale group( DOS),and another 10 rats were used as normal group( Normal). After 7 continuous days of administration,the fecal water content and intestine propulsion rate of each group were detected. HE staining was used to observe the pathological damage of ileum and colon in each group. Immunohistochemistry and Western blot were used to detect aquaporin 3( AQP3) expressions,while the expression levels of the somatostatin( SS) and motilin( MTL) in the ileum of each group were detected by enzyme-linked immunosorbent assay. The results showed that as compared with the model group,the rats in each drug-administered group had increased number of fecal pellets,increased fecal water content,and the increased intestinal propulsion rate( P<0. 01),while the pathological damage of the ileum and colon was significantly reduced; the expression of AQP3 protein was significantly decreased( P<0. 01); the level of MTL was significantly increased and the level of SS was decreased( P<0. 01). All DHS groups showed a good dose-effect relationship,and the same dose treatment effect was equivalent to that of DOS,but it was superior to DNS. Therefore,DHS has a significant therapeutic effect on constipation of spleen Yin deficiency type,and its mechanism may be related to intestinal motility and water-liquid metabolism,with a good therapeutic effect.
Books on Chinese herbal medicines have shown that Dendrobium has the effect of nourishing Yin and reinforcing Yin,usually used for constipation induced by spleen Yin deficiency in clinical application. D. huoshanense,as an independent species among many species of Dendrobium,has no experimental studies about its effects on spleen Yin deficiency-type constipation. The purpose of this experiment was to illustrate the therapeutic effect of D. huoshanense on the constipation of spleen Yin deficiency type in rats,investigate its preliminary mechanism,and compare it with the D. officinale and D. nobile contained in the Chinese Pharmacopoeia to clarify its characteristics. The spleen Yin deficiency model was replicated in 70 rats by the composite factor method,and then the model rats were randomly divided into 7 groups: model group,Liuwei Dihuang Pills group( LWDHP),D. huoshanense high( DHS-H),medium( DHS-M),low( DHS-L) dose groups,D. nobile group( DNS),and D. officinale group( DOS),and another 10 rats were used as normal group( Normal). After 7 continuous days of administration,the fecal water content and intestine propulsion rate of each group were detected. HE staining was used to observe the pathological damage of ileum and colon in each group. Immunohistochemistry and Western blot were used to detect aquaporin 3( AQP3) expressions,while the expression levels of the somatostatin( SS) and motilin( MTL) in the ileum of each group were detected by enzyme-linked immunosorbent assay. The results showed that as compared with the model group,the rats in each drug-administered group had increased number of fecal pellets,increased fecal water content,and the increased intestinal propulsion rate( P<0. 01),while the pathological damage of the ileum and colon was significantly reduced; the expression of AQP3 protein was significantly decreased( P<0. 01); the level of MTL was significantly increased and the level of SS was decreased( P<0. 01). All DHS groups showed a good dose-effect relationship,and the same dose treatment effect was equivalent to that of DOS,but it was superior to DNS. Therefore,DHS has a significant therapeutic effect on constipation of spleen Yin deficiency type,and its mechanism may be related to intestinal motility and water-liquid metabolism,with a good therapeutic effect.
引文
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[10]斯金平,张媛,罗毅波,等.石斛与铁皮石斛关系的本草考证[J].中国中药杂志,2017,42(10):2001.
[11]钱明雪,李胜立,李凡,等. 6种石斛多糖抗亚急性酒精性肝损伤作用的比较[J].中国药学杂志,2015,50(24):2117.
[12]王凯,眭丹娟,王长锁,等. 5种石斛对四氯化碳致小鼠急性肝损伤的保护作用[J].中国中药杂志,2017,42(10):1945.
[13] Pan L H,Li X F,Wang M N,et al. Comparison of hypoglycemic and antioxidative effects of polysaccharides from four different Dendrobium species[J]. Int J Biol Macromol,2014,64:420.
[14]于凡,熊洪艳,梁玲,等.增液汤对便秘模型大鼠血清MTL和SS含量的影响[J].云南中医中药杂志,2017,38(12):61.
[15]王郁金,周永学,谢培,等.硝菔通结方对功能性便秘大鼠排便及胃肠动力的影响[J].中国实验方剂学杂志,2014,20(17):146.
[16] Nobutomo I,Risako K,Kiyoshi S. Aquaporins in the colon as a new therapeutic target in diarrhea and constipation[J]. Int J Mol Sci,2016,17(7):1172.
[17]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.
[2]叶红传,吕小凤,胡苑珂,等.黄德铨教授甘淡滋脾法治疗脾阴虚便秘经验浅析[J].亚太传统医药,2017,13(23):104.
[3] Suares N C,Ford A C. Prevalence of,and risk factors for,chronic idiopathic constipation in the community:systematic review and Meta-analysis[J]. AM J Gastroenterol,2011,106(9):1582.
[4]杜丽东,田金徽,吴国泰,等.基于文献的中药治疗便秘的社会网络分析[J].中国中药杂志,2017,42(2):370.
[5]刘俊兰,于漫,翟美丽,等.脾阴虚证与自由基、能量代谢的相关性及理脾阴正方对其的作用[J].中华中医药学刊,2017,35(4):843.
[6]刘俊兰,于漫,翟美丽,等.理脾阴正方对脾阴虚大鼠能量代谢障碍调节作用[J].辽宁中医药大学学报,2017,19(1):28.
[7]于漫,王彩霞,崔家鹏.脾阴虚证脾失健运大鼠回肠组织中相关蛋白表达差异及其调控机制的实验研究[J].中华中医药学刊,2017,35(7):1726.
[8]肖微,陈鸣,胡瑞连,等.复方植物纤维粉改善人体功能性便秘效果评价[J].慢性病学杂志,2016,17(4):384.
[9]郝菲菲,刘婉青,聂克.连翘对小鼠胃肠动力影响的实验研究[J].山东中医药大学学报,2018,42(4):376.
[10]斯金平,张媛,罗毅波,等.石斛与铁皮石斛关系的本草考证[J].中国中药杂志,2017,42(10):2001.
[11]钱明雪,李胜立,李凡,等. 6种石斛多糖抗亚急性酒精性肝损伤作用的比较[J].中国药学杂志,2015,50(24):2117.
[12]王凯,眭丹娟,王长锁,等. 5种石斛对四氯化碳致小鼠急性肝损伤的保护作用[J].中国中药杂志,2017,42(10):1945.
[13] Pan L H,Li X F,Wang M N,et al. Comparison of hypoglycemic and antioxidative effects of polysaccharides from four different Dendrobium species[J]. Int J Biol Macromol,2014,64:420.
[14]于凡,熊洪艳,梁玲,等.增液汤对便秘模型大鼠血清MTL和SS含量的影响[J].云南中医中药杂志,2017,38(12):61.
[15]王郁金,周永学,谢培,等.硝菔通结方对功能性便秘大鼠排便及胃肠动力的影响[J].中国实验方剂学杂志,2014,20(17):146.
[16] Nobutomo I,Risako K,Kiyoshi S. Aquaporins in the colon as a new therapeutic target in diarrhea and constipation[J]. Int J Mol Sci,2016,17(7):1172.
[17]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.