| BackgroudThe theory of entrails being inerior-exteriorly related is one of the important parts of chinese medicine organ picture theory. And it emphasizes on intercoordination and mutual restriction of entrails being inerior-exteriorly related in physiological state, and interaction and transmission of a disease between organs in pathological state. Also it pays attention to correlated organs during the treatment in order to prevent deterioration and recover ying yang balance and qi xue harmony. The theory of lung and the large intestine being inerior-exteriorly related is the representative of entrails theory. The relationship between lung and intesine is not only embodied by meridian and vessels,meanwhile, which are in a symbiosis inseparable relationship in physiological and pathological state, but also it is confirmed in the long time clinic practice. Nevertheless,whether the structure basis of lung and intestine has the same homology, whether the active compound has specificity, how the cotrol mechanisim works, have no satisfied answer now. Explaining above questions scientifically, revealing biological basis of relationship between lung and large intestinal,and elucidating internal connection and concordance mechanism, could better instruct clinical application.ObjectiveIn the instruction of TCM theory of "the lung and the large intestine being interior-exteriorly related",and following TCM etiology theory, the animal model of intestinal dryness and comsumption of fruid is estabished. Detecting physiological function of pulmonary system and intestine system as the starting point,the related index about the effect of fluid deficiency on physiologic function of the lung and the large intestine were choosed. The study about intervention and mechanism of action of the Fluid-Increasing Decoction for Purgation in rat model was carried out,to elucidate biological basis of physiological and pathological relationship between lung and large intestinal,to reveal their internal connection and concordance mechanism.Methord1.Theory approaching:on the basis of theory of the lung and the large intestine being interior-exteriorly related body, by analyzing the important effect of fluid and its metabolism on physiological function of lung and the large intestine, and on the view of lung qi obstruction caused by intestinal dryness and consumption of fluid an obstruction of fu-qi,the internal mechanism of comorbidity of lung and the large intestine was approached, and their biological basis was analyzed2.Empirical study(1)Modelling test:64 SD rats,200±20g, male and female with each in half, were randomly divided into 4 groups:blank group, water-limited group, complex diphenoxylate group, water-limited +complex diphenoxylate group, each group has 16 rats.When the rats model were established, the alteration of stool was observed. After 7 and 14 days,the water content of the lung and large intestine and pathological sectionwere tested, to establish and evaluate three constipation model as further study basis.(2)Observing effect of Fluid-Increasing Decoction for Purgation on intestinal dryness and consumption of fluid in rat model:60 SD rats,200±20g, male and female with each in half, were randomly divided into 5 groups:blank group, model group, Fluid-Increasing Decoction for Purgation group,Fluid-Increasing Decoction group, western medicine group, each group has 12 rats. The model of intestinal dryness and consumption of fluid was constructed by limiting water and lavaging compex diphenoxylate. After 7 days,the water content of the lung and large intestine, pathological section, lung channel protein land 3, the alteration of VIP,P in the large intestine tissue plasma,were tested.Result1.Modelling(1)observing stool texture and easy and smooth degree of defecation:water-limited,and water-limited add medication administration group:the amount of stool was decreased, fecal pellet became hard and dry, the shape was shorten, discarding with great power and slowly. Medication administration group:most fecal pellet became hard, their surface became dry, sometimes the soft and no formed stool could be seen. Blank group:there is no abmormal in stool texture and defecation.(2)Histopathology observation:blank group were with well-distributed alveoli pulmonis and medium lung interstitium on the 7 and 14th day; the tissue structure of colon was normal,with abundant tomentum and cells lining up in order.Water-limited group:air tube and cilium could not be seen obviously defect.And alveoli pulmonis could not be seen obviously abnormity,the tissue structure of colon was normal,with abundant tomentum on the 7 th day. On the 14 th day, alveolar ectasia and lung interstitial substance broaden, there were some phlegmasia cell in bronchus,.The tissue structure of colon was normal with no obviously pathological changes.Medication administration group:alveoli pulmonis and lung interstitial substance could not be seen obviously abnormity. The tissue structure of colon was normal,with abundant tomentum on the 7 th and 14th day.Water-limited+ medication administration group:alveoli pulmonis and lung interstitial substance could not be seen obviously abnormity and the tissue structure of colon was normal on the 7th day. On the 14 th day, alveolar ectasia and lung interstitial substance broaden, there were some phlegmasia cell in small bronchus, and dropsy cell in alveoli pulmonis.Villus intestinalis were decreased and became short villus intestinalis shape,and lamina propria became thin. And there were eosinophile granulocyte and lympholeukocyte.Under electron microscope, the blank group:on the 7 and 14th day, the tissue structure of lung and colon were normal.The medication administration group:the tissue structure of lung and colon were with no obviously differece.Water-limited group:the tissue structure of lung and colon were with no obviously differece on the 7 th day. On the 14 th day, lung diaphragm were fallen off, lamellar body in endochylema were fallen off.In intestine tissue collagen fibrils were exist.Water-limited+ medication administration group:lamellar body in lung tissue were partly fallen off on the 7th day. On the 14th day, a great quantity of lamellar body in lung diaphragm were fallen off,In intestine tissue collagen fibrils were hyperplasy.(3)The water content in lung and intestine tissue:on the 7 th day, water-limited group and water-limited+ medication had significant difference from blank group in lung and intestine tissue (p<0.01),but medication administration group had no significant difference from blank group(p>0.05);Water-limited group had no significant difference from water-limited+ medication administration group in lung tissue (p>0.05),but has significant difference in intestine tissue (p<0.01).It is indicated that the method of water-limited and water-limited+ dose regimen could cut down the water content in lung and intestine tissue. But this two methods had no significant difference.The water content in lung tissue:on the 14th day, water-limited group and water-limited + medication had significant difference from blank group (p<0.01),but medication administration group had no significant difference from blank group(p>0.05);Water-limited group had difference from water-limited+medication administration group (p<0.05).The water content in intestine tissue:water-limited group and water-limited+medication had significant difference from blank group (p<0.01),but medication administration group had no significant difference from blank group(p>0.05);Water-limited group had no difference from water-limited+medication administration group(p>0.05).It is indicated that the method of water-limited and water-limited+dose regimen could cut down the water content in lung and intestine tissue, but it was more obvious in lung tissue, and there was no significant difference in intestine tissue.2.Observing effect of Fluid-Increasing Decoction for Purgation on intestinal dryness and consumption of fluid in rat model (1)observing stool texture and easy and smooth degree of defecation:model group:the amount of stool was decreased, fecal pellet became dry, the shape was shorten, discarding with great power and slowly. Fluid-Increasing Decoction for Purgation group: there were more fecal pellet than model group, the shape was shorten, sometimes soft.Fluid-Increasing Decoction and western medicine group:fecal pellet became hard, he shape was shorten, discarding with great power and slowly. Blank group:there is no abmormal in stool texture and defecation.(2)Histopathology observationLung:blank group were with well-distributed alveoli pulmonis and medium lung interstitium. Model group:pilus in air tube were with lodging shape, adherence, and defecting lamellar.alveoli pulmonis were engorged and dropsy. Western medicine group:alveolar ectasia,there were great amounts of heterophil granulocyte in lung interstitium. Fluid-Increasing Decoction for Purgation group: there were small amounts of heterophil granulocyte.Enteropathy:blank group:tissue structure of colon was normal, with abundant tomentum. Model group:tomentum were decreased, became short lodging shape,lamina propria became thin, and there were eosinophile granulocyte cells.Fluid-Increasing Decoction and western medicine group:there were eosinophile granulocyte cells and lympholeukocytes.Fluid-Increasing Decoction for Purgation group:alveolar ectasia:only small amounts of eosinophile granulocyte cells.(3)Water contentModel group had significant difference from blank group (p<0.01),it was indicated that model was successful.Lung: Fluid-Increasing Decoction for Purgation group had significant difference from model group (p<0.01),but western medicine and Fluid-Increasing Decoction group had no significant difference from model group(p>0.05);intestine:Fluid-Increasing Decoction for Purgation group and Fluid-Increasing Decoction group had significant difference from model group (p<0.01),but western medicine group had no significant difference from model group(p >0.05).(4)Testing the content of channel protein(AQP1,AQP3)Compared with blank group, the expression of lung AQP1 and intestine AQP3 in model group were decreased(P<0.05).Compared with model group, western medicine and Fluid-Increasing Decoction group AQP1 and AQP3 had no significant difference (p>0.05). Fluid-Increasing Decoction for Purgation group, AQP1 and AQP3 were increased (P<0.05).(5)Testing VIP. P in tissueCompared with blank group, the content of VIP in model group were decreased(P<0.01).Compared with model group, Fluid-Increa sing Decoction for Purgation group, VIP were increased(P<0.05), Fluid-Increasing Decoction group were increased(P<0.05).Compared with blank group, the content of SP in model group were decreased(P<0.01).Compared with model group, Fluid-Increasing Decoction for Purgation group, VIP were increased(P<0.05), Fluid-Increasing Decoction group were increased(P<0.05).Result1.The model of limited water+complex diphenoxylate is more near the clinical expression and basic pathological feature of intestinal dryness and consumption of fluid, and constipation. It is a reliable animal model.2.Water channel protein 1,3 play an important role in body fluid metabolism of lung and large intestine. It contains a low contents when body fluid deficient.3.P, VIP expression in model group is obviously lower than normal control.It maybe the cause of the movement of intestinal tract stepping down.
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