加味桔梗汤对慢传输便秘小鼠模型的影响
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摘要
慢传输型便秘(slow transit constipation,STC)是临床较常见的一类功能性顽固性便秘,主要表现为大便次数减少,便意消失,伴腹胀,其发病机制不清。现代医学的保守治疗对STC疗效有限,严重者最终多外科手术切除全(次全)结肠,但术后频发的粘连性梗阻、腹泻、便秘复发等问题一致困扰着外科医师。
根据其临床表现,STC属于中医学“便秘”范畴,历代医家通过长期的临床观察和实践积累,逐步完善并发展了对便秘的理论认识和辨证治疗,认识到其病位虽在肠,但与肝、脾、肾、肺、胃等脏腑功能失调均密切相关。在中医藏象理论,特别是“肺合大肠”脏腑相关理论指导下,历代医家整体辨治慢传输型便秘临床疗效确切,亦积累了丰富的经验,为防治这种难治性疾病开辟了新思路。“肺病及肠”及“肠病治肺”是“肺合大肠”理论的病理表现形式和实践认知,以其为切入点开展相关机制研究不仅能印证“肺合大肠”理论,亦将为进一步揭示其理论内涵和阐释其生物学机制提供依据。
本课题组在文献研究和理论探讨的基础上,设计动物实验,通过观察加味桔梗汤对慢传输型便秘小鼠模型的影响,探讨STC从肺论治的效应机制,为揭示“肺合大肠”理论的现代生物学机制提供参考。
第一部分文献研究
综述一慢传输型便秘的现代医学研究概括,系统归纳了现代医学有关慢传输型便秘的临床表现、可能发病原因、相关病理机制、治疗进展、研究展望等。综述二中医药治疗慢传输型便秘的研究进展,回顾总结了便秘的中医病因病机、辨证治疗、中医药辨治STC的实验研究及其现代机制；综述三对目前常见的实热型、寒积型、燥结型、阴虚血瘀型、脾虚型、药物性便秘等模型进行了比较和总结,为开展实验研究奠定基础。
第二部分理论研究
基于祖国医学脏腑相关理论,特别是“肺合大肠”理论,结合《伤寒杂病论》肺肠疾病六经辨治相关方证,运用文献学方法,对便秘从肺论治的古今文献进行了整理研究,其中包括辨治原则、证治规律、方药配伍特点等,为今后临床辨治便秘和进行相关科学研究提供了理论依据。
第三部分实验研究
40只小鼠随机分为正常组、模型组、模型给药组及正常给药组,采用自身粪便配合复方地芬诺酯,进行联合灌胃法以建立小鼠便秘模型。模型给药组及正常给药组以加味桔梗汤(桔梗、生甘草、浙贝、紫苑)灌胃,正常组、模型组灌胃蒸馏水,连续7d,观察小鼠的排便情况、推进率及肺肠组织病理学改变,并采用放免法和高压液相色谱法检测各组小鼠肺、肠组织中NKA. VIP和TFF3的含量。
结果发现：与正常组比较,模型组小鼠首粒黑便时间延长,4h排便粒数、4h排便量、肠道炭末推进率均明显减少(P<0.01或P<0.05),肺肠组织中NAK含量均明显升高,VIP含量均明显降低。与模型组比较,模型给药组小鼠首粒黑便时间缩短,4h排便粒数、4h排便重量、肠道炭末推进率增加(P<0.05),肺肠组织中NKA含量明显降低,肺组织中VIP含量明显升高；Spearman简单相关分析显示：肺、肠组织中NKA、VIP、TFF3含量均呈明显相关关系,其相关系数分别为0.448、0.503、0.423。以上结果提示,采用自身粪便与复方地芬诺酯灌胃法制备的小鼠复合便秘模型,存在明显肠道传输功能障碍；宣肺中药加味桔梗汤可促进胃肠蠕动、改善便秘小鼠肠道传输功能,其效应机制可能与调节肺、肠组织中神经肽NKA、VIP的含量有关,为进一步从神经肽角度探讨“肺合大肠”理论提供了佐证。
Slow transit constipation is a clinically common functional intractable constipation, it's main symptom is fewer defecate, no feeling of defecate and with abdominal distension, and the pathogenesis is not clear. The effect of Modern medicine conservative cure for STC is limited and the most serious situation is to cut the all colon, but the adhesion obstruction、diarrhea、 constipation recurrence are still bother the surgeon.
According to the clinical symptom, STC is belong to the category of constipation in Chinese medicine, the famous historical doctors gradually developed a perfect theory, diagnosis and treatment to cure constipation with the long term clinical observation and practice. They knew that although the location is at large intestine, it also has much to do with the imbalance function of liver、spleen、kidney、lung and stomach. In Chinese medicine theory, especially guided by the theory of lung and large intestine, historical doctors got good performance in treating STC and also accumulated rich experience and got a new thinking way to cure this difficult disease. The lung disease transiting to large intestine and treating lung to cure large intestine disease are also according to "The lung and large intestine theory". We also can develop related research, represent the connotation of theory and explain the biology mechanism to improve "The lung and large intestine theory"
With the research and theory discussion, our group design animal experiment to observe augmented Jie geng decoction, having influence on the STC mouse pattern, and to discuss the effect function of curing from lung to represent "The lung and large intestine theory'
The First part of the literature research
Review Ⅰ:The modern medical study of STC can be coucluding:summed up the modern medical clinical presentation about STC, the reasons of the disease upset,related pathological mechanisms,the advances for the treatment and the reasearch propects...etc.
Review Ⅱ:The study of in Chinese STC medical therapy progress:Review and summurize the Chinese medical causes and mechanisms,the treatment of the symdrom differenationmthe experimental study of STC's the traditional chinese medical diagonsis and treatment,and it's modern mechanism.
Review III:Until now,the regular types about the sthenia-heat type,the cold accummulation type,the dry constipation type,the yin deficiency and blood stasis type,the weakness of the spleen,and the drug-induced constipation model were compared and summerized. To form the foundation to carry out the experimental research.
The Second part of theoretical research
The visicera-associated theory based on the traditional Chinese,especially "The lung associ-ated with large intestine"theory. combined with the six meridians disease treatment, using the methods of philology, constipation governance from the lungs on the ancient and modern litera-ture finishing the studies, including Treating the principle of the law of Treatment Recipe compat- ibility features for future clinical Treating constipation and related scientific research provides a theoretical basis.
Third part experimental research
40mice were randomly divided into normal control group, model group, treating group, and normal treating group. The mice model of constipation was established by using Compound Diphenoxylate. The model group and the normal control group were given intragastrically the distilled water, and other groups were given the Augmented Jiegeng Decoction(Jiegeng,Gan cao,Zi wan,and Bei mu) intragastrically for7days. Detecting transmission function of intestine, lung and intestine pathology,the promoting rate of ink,and the content of NKA,VIP,and TFF3in lung and intestine tissue.
Results： Compared with the normal control group, the time of the first black feces, the faeces quantity within4hours, and the promoting rate of ink were significant difference in model group (P<0.01, or P<0.05); compared with the model group, there were significant improvement in treating group(P<0.05). The content of NKA in lung and intestine tissue is decreased obviously,and the content of VIP in lung tissue is increased obviously. Spearman simple correlation analysis shows that：There are significant correlations between the content of NKA,VIP,and TFF3.The Correlation coefficient are0.448,0.503,and0.423.The result suggested that,there is intestinal dysfunction of transmission in mice constipation model.The facilitating lung function of the Augmented Jiegeng Decoction can enhance the improving in transmission function of intestine in mice with constipation.The possible effect mechanism is regulating neuropeptides NKA,VIP in lung and intestine tissue.The result provide the evidence for further explore to"lung associated with large intestine theory"
根据其临床表现,STC属于中医学“便秘”范畴,历代医家通过长期的临床观察和实践积累,逐步完善并发展了对便秘的理论认识和辨证治疗,认识到其病位虽在肠,但与肝、脾、肾、肺、胃等脏腑功能失调均密切相关。在中医藏象理论,特别是“肺合大肠”脏腑相关理论指导下,历代医家整体辨治慢传输型便秘临床疗效确切,亦积累了丰富的经验,为防治这种难治性疾病开辟了新思路。“肺病及肠”及“肠病治肺”是“肺合大肠”理论的病理表现形式和实践认知,以其为切入点开展相关机制研究不仅能印证“肺合大肠”理论,亦将为进一步揭示其理论内涵和阐释其生物学机制提供依据。
本课题组在文献研究和理论探讨的基础上,设计动物实验,通过观察加味桔梗汤对慢传输型便秘小鼠模型的影响,探讨STC从肺论治的效应机制,为揭示“肺合大肠”理论的现代生物学机制提供参考。
第一部分文献研究
综述一慢传输型便秘的现代医学研究概括,系统归纳了现代医学有关慢传输型便秘的临床表现、可能发病原因、相关病理机制、治疗进展、研究展望等。综述二中医药治疗慢传输型便秘的研究进展,回顾总结了便秘的中医病因病机、辨证治疗、中医药辨治STC的实验研究及其现代机制；综述三对目前常见的实热型、寒积型、燥结型、阴虚血瘀型、脾虚型、药物性便秘等模型进行了比较和总结,为开展实验研究奠定基础。
第二部分理论研究
基于祖国医学脏腑相关理论,特别是“肺合大肠”理论,结合《伤寒杂病论》肺肠疾病六经辨治相关方证,运用文献学方法,对便秘从肺论治的古今文献进行了整理研究,其中包括辨治原则、证治规律、方药配伍特点等,为今后临床辨治便秘和进行相关科学研究提供了理论依据。
第三部分实验研究
40只小鼠随机分为正常组、模型组、模型给药组及正常给药组,采用自身粪便配合复方地芬诺酯,进行联合灌胃法以建立小鼠便秘模型。模型给药组及正常给药组以加味桔梗汤(桔梗、生甘草、浙贝、紫苑)灌胃,正常组、模型组灌胃蒸馏水,连续7d,观察小鼠的排便情况、推进率及肺肠组织病理学改变,并采用放免法和高压液相色谱法检测各组小鼠肺、肠组织中NKA. VIP和TFF3的含量。
结果发现：与正常组比较,模型组小鼠首粒黑便时间延长,4h排便粒数、4h排便量、肠道炭末推进率均明显减少(P<0.01或P<0.05),肺肠组织中NAK含量均明显升高,VIP含量均明显降低。与模型组比较,模型给药组小鼠首粒黑便时间缩短,4h排便粒数、4h排便重量、肠道炭末推进率增加(P<0.05),肺肠组织中NKA含量明显降低,肺组织中VIP含量明显升高；Spearman简单相关分析显示：肺、肠组织中NKA、VIP、TFF3含量均呈明显相关关系,其相关系数分别为0.448、0.503、0.423。以上结果提示,采用自身粪便与复方地芬诺酯灌胃法制备的小鼠复合便秘模型,存在明显肠道传输功能障碍；宣肺中药加味桔梗汤可促进胃肠蠕动、改善便秘小鼠肠道传输功能,其效应机制可能与调节肺、肠组织中神经肽NKA、VIP的含量有关,为进一步从神经肽角度探讨“肺合大肠”理论提供了佐证。
Slow transit constipation is a clinically common functional intractable constipation, it's main symptom is fewer defecate, no feeling of defecate and with abdominal distension, and the pathogenesis is not clear. The effect of Modern medicine conservative cure for STC is limited and the most serious situation is to cut the all colon, but the adhesion obstruction、diarrhea、 constipation recurrence are still bother the surgeon.
According to the clinical symptom, STC is belong to the category of constipation in Chinese medicine, the famous historical doctors gradually developed a perfect theory, diagnosis and treatment to cure constipation with the long term clinical observation and practice. They knew that although the location is at large intestine, it also has much to do with the imbalance function of liver、spleen、kidney、lung and stomach. In Chinese medicine theory, especially guided by the theory of lung and large intestine, historical doctors got good performance in treating STC and also accumulated rich experience and got a new thinking way to cure this difficult disease. The lung disease transiting to large intestine and treating lung to cure large intestine disease are also according to "The lung and large intestine theory". We also can develop related research, represent the connotation of theory and explain the biology mechanism to improve "The lung and large intestine theory"
With the research and theory discussion, our group design animal experiment to observe augmented Jie geng decoction, having influence on the STC mouse pattern, and to discuss the effect function of curing from lung to represent "The lung and large intestine theory'
The First part of the literature research
Review Ⅰ:The modern medical study of STC can be coucluding:summed up the modern medical clinical presentation about STC, the reasons of the disease upset,related pathological mechanisms,the advances for the treatment and the reasearch propects...etc.
Review Ⅱ:The study of in Chinese STC medical therapy progress:Review and summurize the Chinese medical causes and mechanisms,the treatment of the symdrom differenationmthe experimental study of STC's the traditional chinese medical diagonsis and treatment,and it's modern mechanism.
Review III:Until now,the regular types about the sthenia-heat type,the cold accummulation type,the dry constipation type,the yin deficiency and blood stasis type,the weakness of the spleen,and the drug-induced constipation model were compared and summerized. To form the foundation to carry out the experimental research.
The Second part of theoretical research
The visicera-associated theory based on the traditional Chinese,especially "The lung associ-ated with large intestine"theory. combined with the six meridians disease treatment, using the methods of philology, constipation governance from the lungs on the ancient and modern litera-ture finishing the studies, including Treating the principle of the law of Treatment Recipe compat- ibility features for future clinical Treating constipation and related scientific research provides a theoretical basis.
Third part experimental research
40mice were randomly divided into normal control group, model group, treating group, and normal treating group. The mice model of constipation was established by using Compound Diphenoxylate. The model group and the normal control group were given intragastrically the distilled water, and other groups were given the Augmented Jiegeng Decoction(Jiegeng,Gan cao,Zi wan,and Bei mu) intragastrically for7days. Detecting transmission function of intestine, lung and intestine pathology,the promoting rate of ink,and the content of NKA,VIP,and TFF3in lung and intestine tissue.
Results： Compared with the normal control group, the time of the first black feces, the faeces quantity within4hours, and the promoting rate of ink were significant difference in model group (P<0.01, or P<0.05); compared with the model group, there were significant improvement in treating group(P<0.05). The content of NKA in lung and intestine tissue is decreased obviously,and the content of VIP in lung tissue is increased obviously. Spearman simple correlation analysis shows that：There are significant correlations between the content of NKA,VIP,and TFF3.The Correlation coefficient are0.448,0.503,and0.423.The result suggested that,there is intestinal dysfunction of transmission in mice constipation model.The facilitating lung function of the Augmented Jiegeng Decoction can enhance the improving in transmission function of intestine in mice with constipation.The possible effect mechanism is regulating neuropeptides NKA,VIP in lung and intestine tissue.The result provide the evidence for further explore to"lung associated with large intestine theory"
引文
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