| BackgroundIn acupuncture clinical practice, doctors often encounter patients with chronic atrophic gastritis, which some patients are usually accepted and the efficacy of various treatments after less satisfied, come to seek help acupuncturist. It is a common disease, but its etiology complicated, there is no uniform plan to follow, and the effects of various treatments unsatisfactory, thus becoming one of the common clinical difficult disease. In acupuncture clinical practice, we found that acupuncture for the treatment of this disease with more satisfactory results, and many randomized controlled clinical trials have been reported in the literature confirms that acupuncture treatment of chronic atrophic gastritis with good effect, but its intrinsic mechanism is not clear. Therefore, the design rigorous animal experiments to explore the mechanism of acupuncture treatment of this disease is particularly necessary. NF-κ B signaling pathway is one of the three major pathways of various inflammatory responses, but also affect the process of apoptosis inflammation important factor. To explore the molecular mechanism of acupuncture treatment of chronic atrophic gastritis, based on existing research results and mentor in a lot of literature on the basis of inspection, we hypothesize:The mechanism of acupuncture to treat chronic atrophic gastritis may be that acupuncture can restrain the activation of NF-κ B-way signal transduction systems, then control the progress of inflammation, reduce the inflammatory reaction, regular the gene expression of anti-apoptosis, alleviate cell apoptosis of atrophic gastritis and then protect gastric mucosa. Therefore, this study hopes to study acupuncture impact on the gene expression of TLR4、IL-1RI、TAK1、IKKβ、IκB、NF-κ B、IL-8IL-1R, TNFR and TUNEL in the NF-κ B-way signal transduction on the rat model with chronic atrophic gastritis. On the one hand to observe the curative effect of acupuncture on chronic atrophic gastritis, on the other hand to explore the effect of acupuncture on NF-κ B-way signal transduction, then to enrich the relative theory of acupuncture to treat chronic atrophic gastritis, enhance the value and use value to guide clinical and provide new clue to prevent precancerous lesions of chronic atrophic gastritis by acupuncture.ObjectiveBy establishing a rat model of chronic atrophic gastritis, investigate NF-κB signaling pathway in TLR4, IL-1RI, TAK1, IKKβ, I κ B, NF-κB, IL-8, IL-1R, TNFRI and TUNEL chronic atrophic Expression and significance of gastritis in rats, acupuncture effects on the expression of these indicators, and to explore acupuncture on NF-κB transduction system to provide theoretical and experimental evidence for acupuncture treatment of chronic atrophic gastritis.MethodsOptional80weighing180-200g of male and female SD rats were randomly established with the control group25, CAG-made module55. Blank control group of normal feeding. Modeling modules made using the following method for preparing a rat model of chronic atrophic gastritis for:Configure a solution concentration of MNNG lg/L, the dark cryopreservation; when diluted with distilled water to100μg/ml, for rats in drinking, in conjunction with irregular diet (ie enough to feed one day, fasting one day, alternating between the two). Rat model group were killed in a randomized, light microscope after gastric pathology confirmation to establish successful, the module will be made using a random number table remaining rats were divided into model group and acupuncture group. This study successfully replicated the model, the model group21rats, acupuncture group was22(process modeling died six rats were randomly sacrificed6), then carry out the experiment. Control group of rats, normal feeding interventions are not applied until the end of the experiment. Model rats, normal feeding interventions not applied until the end of the experiment. Acupuncture rats, normal feeding, and applying acupuncture interventions until the end of the experiment. Acupuncture group interventions: the30th one-inch stainless steel needle3acu-points Zusanli (ST36), Zhongwan (RN12) and Spleen (BL24). Stabbing desired depth, get the arrival-qi and keep needles for15mins. Treatment once a day,10days for a course of treatment, the total treatment needs6courses.After treatment completed, all rats were sacrificed conducted to detect the observed indicators:1. Gastric pathology observed;2. Lesion index;3. ELISA detection of TNF-α, TNFRI and IL-6;4. Immunohistochemistry IL-1RI this signaling pathway, TAK1, TLR4, IKK, I κ B, NF-κB and IL-8cytokine;5.Gastric mucosal cell apoptosis index;6. Gastric mucosal cell ultrastructure.Achi evement1. In12weeks, randomly kill a rat, if the gastric pathological changes could be observed by light microscopy, the result of building model was successful and then all the rats enter the treatment phase. No rats died in the course of therapeutic intervention; ultimately included in the study rats were66,23blank group, model group21, the acupuncture group22.2. Observation the gastric pathological changes:in the blank group, the changes of gastric mucosa are normal; in the model group, the changes of gastric mucosa are in line with the model group pathological diagnostic criteria of chronic atrophic gastritis; In the acupuncture group, the gastric mucosa of rat had varying degrees of improvement.3. The rats in model group compared with the rats in the blank group on the gastric mucosal injury index, serum CRP, TNF-α, TNFRI and IL-6was increased, the difference was statistically significant (P<0.05or P<0.01); Immunohistochemical optical density relatively prompt, model group IL-1RI, TAK1, TLR4, IKKβ, IκBα, NF-κ B (p65) and IL-8were higher than the control group of rats, the difference was statistically significant (P<0.05or P<0.01); these indicators, the acupuncture group compared with model group, the difference was statistically significant (P<0.05or P<0.01), but little difference with the control group (P>0.05).4. The immunohistochemistry expression of IL-1RI, TAK1, TLR4, IKKβ, IκBα, NF-κ B (p65) and IL-8of gastric mucosa of rats in each group:the blank control group is faint yellow, the expression is low; the model group is brown, the expression is high; the color in the acupuncture group is between blank control group and model group, but close to blank control group, so its expression is moderate.The apoptotic index of gastric mucosa cell of rats in model group was significantly higher than the control group (P=0.01); in the acupuncture group, the index of gastric mucosa cell of rats compared with the model group decreased significantly (P<0.05), and the gap is not large with the blank control group (P>0.05).6. Observation on the ultrastructure of gastric mucosal cell:in the blank control group, the structure of organelle is normal on the gastric mucosa chief cell and parietal cell of rats; in the model group, the mirror image structure of organelle was obviously destroyed on the gastric mucosa chief cell and parietal cell of rats.Conclusion1. Application of free drinking solution100μg/ml concentration of MNNG and abnormal of starvation and full method can build the rat model of chronic atrophic gastritis successfully in12weeks.2. Applying above model building method, NF-κB activation signal transduction systems can be detected on the rat model of chronic atrophic gastritis, mainly for, TNF-α, IL-6, IL-8and inflammatory pathways on specific receptors, cytokine TNFRI, TAK1, IL-1R1, TLR4high expression; Meanwhile, the core factor of IKKβNF-κ B signal transduction pathway, IκBα, NF-κ B (p65) expression level compared with the rats in the blank group was significantly increased; in addition, NF-κ B signal transduction pathway regulated by the product of TNF-α, IL-6, IL-8cytokine expression was significantly increased; The apoptotic index of gastric mucosa cell of rats was significantly higher3. Acupuncture can uni-direction restrain the activation of NF-κ B-way signal transduction systems, mainly for, the expression of the IKKP, I κ Bα, NF-κ B (p65) is lower than that of model group and close to the normal group; its upstream cytokines TNFRI, TAK1, IL-1R1, TLR4and its downstream products of TNF-α, IL-6, IL-8are also lower than that of model group; apoptotic index of gastric mucosa cell of rats was decreased and close to the normal group.In conclusion, acupuncture for the rat model with chronic atrophic gastritis have regulating effect on the NF-κ B signaling pathway. Mainly for, acupuncture can uni-direction restrain the activation of NF-κ B-way signal transduction systems and effect on the NF-κ B into the nucleus regulating gene transcription process, by affecting the NF-κ B-induced anti-apoptotic protein coding genes promoting cell survival and induction of the gene encoding the expression of anti-oxidative to restrain the process of transduction system, such as cell apoptosis and so on to protect the gastric mucosa cell. Acupuncture has obviously curative effect on the model rat with chronic atrophic gastritis, which be realized probably by regulating the NF-κ B signal transduction system of the rat model with chronic atrophic gastritis. |
PDF Full Text Request