| ObjectiveIt is found in recent years that there are sweet taste receptors and signaling molecules in the gastrointestinal tract closely related to the nutrient metabolism. TCM has the theory from ancient time that spleen is reflected as sweet in flavor. Related reports point out that the spleen is closely related with intestinal metabolism of nutrients. At the mean time, it can regulate the appetite and affect the energy balance by tonifying spleen and stomach. The report is in accordance with the transduction function of intestinal sweet taste signaling molecules, and also corresponding to the TCM theory. Therefore, from the level of transduction function of intestinal sweet taste signaling molecules to explore the essence of the spleen function, is helpful for revealing the connotation of the TCM theory that spleen is reflected as sweet in flavor. And studying the relationship between spleen and intestinal sweet taste receptors and key signaling molecules will also help to clarify the TCM treatment for functional diarrhea through the spleen, which has practical guiding significance.The spleen is corresponding to earth of the five elements, which is named Jialian (means sowing and collecting). Everything is born in the earth, and everything dies in the earth. Spleen and stomach are the acquired foundation, which have inoculation and nourishing functions for all things. In the five zang-organs, the spleen is corresponding to the sweet of the five flavors, with the function presenting as the inoculation and nourishing of "earth". Everything in human body depends on the transportation, transformation and essence distribution of the spleen and stomach for nourishing. Regardless of tonifying Yin, Yang, Qi or blood function, sweet medicine needs the role of the spleen and stomach to achieve. Spleen governs transportation and transformation of dampness. Spleen Yang deficiency is unable to transport dampness, thus the water dose not move, and the dampness gathers to form fluid retention. So in clinical, phlegm and fluid retention, and dampness-heat of spleen and stomach are mainly treated according to the principles of tonifying spleen and stomach, clearing dampness and promoting dieresis. It is easy to find that many of the damp-clearing drugs are sweet, with the functions of tonifying spleen, clearing dampness and promoting dieresis. This is also the embodiment of "sweet flavor attributes to the spleen and spleen is reflected as sweet in flavor".Functional diarrhea (FDi) is a common functional bowel disorder (FBD) in clinics. Stimulated by some pathogeneses, gastrointestinal movement and absorption functions are made disordered, thereby impacting its secretion and absorption functions. Due to incomplete digestion, its secretion functions are accelerated while with a decreased absorption capacity, leading to runny stools, frequent bowel movements as well as continuous or repeated loose stools without stomachache or discomfort, thereby forming functional diarrhea. At present, it is still unclear for etiology and pathogenesis of functional diarrhea. In terms of therapeutic method, effective specific therapeutic regimens are still unavailable. However, Chinese traditional medicine has certain advantages in treatment of FDi, for instance, curative effects, less side effects, effective remission of patients’ clinical symptoms, and improvement to patients’life quality.Functional diarrhea belongs to the category of "diarrhea" in traditional Chinese medicine (TCM), with the main pathological lesions in the spleen, stomach and intestines. Its causes include exogenous pathogenic factors and disorder of internal organs. But the most important reason lies in the spleen itself. The old saying goes "Dampness induces the five kinds of diarrhea" and "no dampness no diarrhea". The spleen prefers dryness to dampness. When exogenous pathogenic factor invading the human body combined with dampness pathogenic factor, it will affect the transportation and transformation function of the spleen and stomach, thus causing imbalance of the ascending-descending, poor transportation and transformation, disorder of the conduction of large intestine and finally the diarrhea. Internal causes of the disease include the weakness of the spleen and the dysfunction of the spleen and the stomach, so the spleen loses the transportation and transformation function, causing disorder of the ascending and descending of Qi transportation. As a result, the clear Qi and the foul Qi cannot be separated, which causes the conduction out of control, thus leading to diarrhea. The dampness and the spleen deficient influence each other during the onset and the pathological changes. Spleen deficient and excessive dampness are the key to diarrhea. Spleen deficient is the root cause and excessive dampness is the manifestation. The lesions are in the intestine but the main responsible organ for the disease is the spleen. In addition the disease is also closely related with the liver and the kidney. So, functional diarrhea should be treated through the spleen.Research objevtive:Experimental section:Physiological and pathological (spleen deficiency) models are established at rats for case-control study. Clinical section: Spleen deficiency patients with intestinal sweet taste transduction related disease ((functional diarrhea) are selected as the subjects. The patients are received tonifying spleen treatment, and detected for expressions of the first taste receptors family (T1R1and T1R2and T1R3) of duodenal mucosa taste pathway and the key signal molecules G a gust and TRPM5well as immunohistochemical quantitative Gα gust before and after the treatment. It is attempted to explore the essence of the spleen function from the intestinal sweet taste signaling molecules and metabolism so as to reveal the relationship the connotation of the TCM theory that spleen is reflected as sweet in flavor. And through confirmation on the close relationship between the spleen, intestinal sweet taste signaling pathways and metabolism, the mechanism of TCM treatment for functional diarrhea through the spleen is revealed.Research methods:1Observation on clinical efficacyOutpatients from Lingnan Consultant Outpatient of the First Affiliated Hospital of Guangzhou TCM University are selected as the research objects. The observation is from March2012to October2013. The patients conforming to the syndromes are included. The diagnosis of western medicine implemented is strictly accordance with the standards;20cases are selected for each syndrome of the functional diarrhea. All the subjects need to sign a consent form. Patients with spleen and stomach deficiency are given prescription I, and patients with dampness-heat of spleen and stomach are given prescription Ⅱ, with20ml each time and3times daily for4weeks. It is compared for the clinical efficacy of primary and secondary TCM symptoms before and after the treatment.2Clinical experimental researchesGastroscope (GIF-XQ260, Olympus Company) is adopted. Two pieces of duodenal mucosal tissues are taken before and after treatment under gastroscope, and then preserved in liquid nitrogen. Western blot and QPCR method are used to detect the expressions of the first taste receptors family (T1R1and T1R2and T1R3) of duodenal mucosa taste pathway and the key signal molecules Gα gust and TRPM5well as immunohistochemical quantitative Gα gust of the two groups of patients with functional diarrhea before and after the treatment. The patients or their families should be explained the purpose before sampling, and only they agree and sign the "informed consent" then they can draw the materials.3Animal experimental researchA total of60SD rats (SPF, male:female=1:1) are provided by Laboratory Animal Center of Guangzhou TCM University (certificate number: SCXK(Yue)2013-0020). The rats (body weight:160-200g) are randomly divided into5groups:spleen deficiency model group (SD-M group), rats with spleen deficiency and natural recovery group (SD-NR group), rats with spleen deficiency and given treatment group (SD-T group), normal control group (NC group) and kidney deficiency control group (KD-C group). SD-M group is given decoction of bark of magnolia, immature bitter orange, and rheum officinale. The rats are given gavage every other day, with feeding on the alternate day for7weeks for modeling. The feeding mode of the SD-NR group is same as the SD-M group for3weeks, and then it changes into the normal diet for4weeks. The feeding mode of the SD-T group is same as the SD-M group for3weeks, and then it changes into gavage of prescription I everyday for4weeks. The NC group is fed normally. The KD-C group is fed adaptively for3weeks, and then given gavage of2ml of2%suspension of adenine (2g/100ml NS) for4weeks.7weeks after modeling, the rats are scarified and dissected. Duodenal tissues are taken and preserved in liquid nitrogen. Western blot and QPCR method are used to detect the expressions of the first taste receptors family (T1R1and T1R2and T1R3) of duodenal mucosa taste pathway and the key signal molecules Gα gust and TRPM5well as immunohistochemical quantitative Ga gust of rats in each group.Research results:1Observation on clinical efficacyClinical studies have shown that in the corresponding spleen and stomach deficiency group and dampness-heat of spleen and stomach group, prescription Ⅰ and Ⅱ can obviously improve the main symptoms including stool property and stool frequency, and can effectively improve the secondary TCM syndromes of patients with functional diarrhea, such as slow statement with fatigue, loss of appetite and borborygmus, and no adverse reactions occur during the treatment.2Clinical experimental researchAfter treatment, expressions of G a gust and TRPM5are both increased in the two groups of patients with functional diarrhea. In spleen and stomach deficiency group, the differences of T1R1, T1R2, T1R3, Ga gust and TRPM5are significant before and after the treatment (P<0.01); After treatment, the expressions of the first taste receptors family (T1R1and T1R2and T1R3) and the key signal molecules Gα gust and TRPM5in spleen and stomach deficiency group and dampness-heat of spleen and stomach group are compared, and the differences are statistically (P<0.01).3Animal experimental researchExperimental studies show that expressions of gestation receptor cells T1R1, T1R2, T1R3,. G a gust and TRPM5in the SD-T group are significantly higher than normal control group (P<0.01); The expressions of gestation receptor cells T1R1, T1R2, T1R3, G a gust and TRPM5in the SD-NR group, SD-M group and KD-C group are all significantly lower than that of the SD-T group (P<0.01).Conclusions①Functional diarrhea can be treated with the spleen theory.②The spleen strengthening method results in an obvious improvement to clinical symptoms of a FDi patient.③The spleen strengthening method can influence the manifestation of taste receptors and key signal molecules in intestinal tracts and interfere the transduction of T1Rs-Gα gust/PKCβ2taste channels of duodenal mucosa.④The pathogenesis of spleen-deficiency syndrome is associated with abnormal manifestation of taste receptors and key signal molecules in intestinal tracts.⑤The spleen strengthening method interferes the transduction of taste channels and recovers the function of the spleen governing movement and transformation by influencing taste receptors in intestinal tracts and signal molecule manifestation, which is consistent with the theory of "the spleen in the taste is sweet" in traditional Chinese medicine. |
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