| Objective:Inter-functional dyspepsia interstitial cells of Cajal (ICC) as a model to study moxibustion Zusanli SD rat serum for interstitial cells of Cajal (Interstitial cells of cajal, ICC) in the inositol triphosphate receptor (Inositol1,4,5-trisphosphate receptor, IP3R) and ryanodine receptors influence (Ryanodinereceptor, RyR) expression, and exploration of the intracellular Ca2+regulating mechanisms, revealing Zusanli moxibustion treatment of functional dyspepsia regulate gastrointestinal mechanism of action sports。Method:1. Establishment of an animal model of functional dyspepsia:a method based on traditional Chinese medicine theory and the theory of the etiology of FD simulation animal model described in the literature dyspepsia, using the rat tail clamp stimulation caused by the line of the fight modeling method of modeling.Modeling7days, and36rats were randomly divided into three groups and fed with normal saline group tied bondage, tied bondage model normal saline group, moxibustion Zusanli normal saline group, a total of lOd,10d after the preparation of the corresponding serum。2. Separation SD rat intestinal ICC were primary cultured under sterile conditions, and method used repeatedly adherent cells were purified by morphology and immunofluorescence study determined that cell identification, animal models with functional dyspepsia serum establish a "functional dyspepsia ICC model"3.by using reverse transcription polymerase chain reaction (RT-PCR) technique for the detection of interstitial cells of Cajal IP3R, RyR and the content of RNA, micro concentration of Ca2+cells, the detection of changes in Cajal plate method, expression of stromal cells in IP3R, RyR protein was detected by Cajal Western-Blot method,so as to reveal the moxibustion at Zusanli in treatment of functional dyspepsia adverse effect mechanism in the regulation of gastrointestinal motility. Result:1. ICC morphology and immune of luorescence identification:ICC inverted microscope observation of the morphological changes:in vitro ICC has special morphological characteristics. Primary ICC were inoculated in culture flask orplate,24h can be at tached to the wall deformation, triangle or fusiform cell, can have a few short processes. Third days cells formed long neurite neurite like.7days later, ICC can extend two level and three level neurites, can beconnected with each other to form a complex network structure, this kind of connection is the structural basis for itsslow wave function. Immun of luorescence identification:To observe the effect of ICC on surface of C-kit was positive in immunofluorescence microscopy, cell body and neuritis were part of a long process of cell coloring.The establishment of functional dyspepsia model:ICC cells by morphological observation, functional dyspepsia Model cell morphology changed obviously: the cells became flat, synaptic connected closely, cell gap is smaller,the network connection is not obvious.2. Tests showed objective indicators of functional dyspepsia model compared with normal ICC, changed:functional dyspepsia model within cells, reduce the level of external Ca2+concentration (P<0.05)。3. The results of Western-Blot assay showed that, compared with the normal cultured interstitial cells of Cajal, FD in serum of rats after the intervention of Cajal interstitial cells of IP3R and RyR proteinexpression were decreased(P<0.01); and the level of serum FD in rats after intervention of interstitial cells of Cajal compared to FD expression of moxibustion at Zusanli point in rats serum intervention interstitial cells of Cajal in IP3R and RyR protein were significantly increased (P<0.01).4. The results of RT-PCR assay showed that, compared with the normal cultured interstitial cells of Cajal rats, serum FD content of dry prognosis of interstitial cells of Cajal IP3R, RyR m RNA were decreased (P<0.01); and the level of serum FD in rats after intervention of interstitial cells of Cajal compared to FD rats serum moxibustion Zusanlicontent of dry prognosis interstitial cells of Cajal IP3R and RyR Mrna were increased (P<0.01).Conclusion:1.primary cell cultures ICC success:be successfully isolated and culture d cells of Cajal from neonatal SD rat intestinal tissue.2.functional dyspepsia ICC cell model success:After the serum was added to normal functional dyspepsia model of ICC, ICC successfully copi ed cell model of functional dyspepsia.3.FD moxibustion Zusanli serum on interstitial cells of Cajal, external Ca2+concentration and IP3R, RyR protein expression and abnormal chang es m RNA content has regulatory role.
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