| Objective: pathogenic cold and dampness is common pathogenic cold dampness pathogen. Cold, wet evil for evil, the most easily damage yang.From the perspective of the theory of traditional Chinese medicine of exogenous cold dampness pathogenic cause of disease mechanism and treatment principle of the external sense disease Alpine Research is of great significance. This study intends to through the establishment of exogenous cold dampness model and investigate the pathogenic mechanism of exogenous cold dampness spleen injury and scattered Quhan wet foreign feeling cold therapeutic effect.method:Theoretical research:Consulting relevant literature, summarizes the pathogenic characteristics of pathogenic cold and dampness, investigate exogenous pathogenic dampness spleen injury pathogenic pathways, pathogenesis and treatment.experimental study:45 SPF grade Kunming mice were randomly divided into 3 groups according to their weight after 1 week: normal control group(normal group): 15 Kunming mice, normal room temperature(15-20 ℃) feeding. Exogenous cold dampness group(hereinafter referred to as model group): 15 Kunming mice,daily for 7 consecutive days at 12:00-16:00 the mouse was placed in a temperature for(4 ±0.5) ℃ and the humidity of 90% ±2 artificial climate box feeding 4h, the rest of the time according to the conventional room temperature feeding. According to 0.13ml/10 g standard, normal saline was administered to the stomach every day. Medication group(hereinafter referred to as the treatment group): 15 Kunming mice, daily for 7 consecutive days at12:00-16:00 the mouse was placed in a temperature for(4 ± 0.5) ℃ and the humidity of 90% ±2 artificial climate box feeding 4h, the rest of the time in room temperature conventional breeding, according to the human and mouse body surface area conversion, according to the standard weight and0.13ml/10 g every day give addition and subtraction renal with soup gavage.After taking the blood animal survival were killed by cervical dislocation.Collection of mouse colonic feces under sterile conditions, and the small intestinal tissue of mice. Objective To observe the survival state of mice,analysis changes in body weight of mice, plate count of bacteria in the gut, the histopathological changes of small bowel pathology structure, staining was used to observe the small intestine of AQP-4 PAR-2, MAPKP38 protein expression by immunohistochemistry, using the bio plex hanging liquid chip technology in the detection of the serum levels of IL-4, IL-10, IL-6, IL-11,TNF, IFN- cytokine expression, to observe the intestinal AQP-4, PAR-2,MAPKP38, NF- KB, My D88, TLR4 Protein Expression by immunoblotting(Western blotting).Resultstheoretical discussionExogenous cold dampness pathogenic pathways analysis: pathogenic cold, wet evil body and pathogenic, the symptoms of the disease alpine, alpine guest list,aversion to cold, cold limbs; resistance on the meridian, cold pain in the joints of the whole body edema, hypertonicity; straight cold of the spleen, stagnation of Qi, see moisten diarrhea, vomiting, abdominal distension full. Cold evil guest, block of Qi, blood and body fluid operation can cause water wet, so that the original pure cold syndrome into cold dampness syndrome; dampness blockage Yang with the passing of time, make Yang injury, resulting in lack of Yang, Yang deficiency cold or body Yang feel wet evil also makes simple dampness, into cold dampness syndrome.Analysis of the pathogenesis of exogenous pathogenic dampness:Interaction of exogenous cold dampness, cold injury of the spleen yang,influence of spleen function; cold dampness pathogen further damage to kidney yang, kidney yang deficiency, unable to warm Spleen Yang, thereby increasing the cold; cold dampness pathogen further role in the human body,damage the Yang and Yin and Yang obstruction, camp Wei discord, surface Yang deficiency, warm and nourish the disorder of Qi; cold dampness pathogen interaction in the human body, easy to make the gas machine not free, affect the spleen and stomach qi and pathogenic dampness obstructing the Sanjiao channel disadvantage.experimental resultsGeneral condition of mice were observed: normal control group of mice lips ruddy, ears, tail is pale pink, hair smooth and shiny, normal food intake, body weight increased gradually, sensitive reaction; model group hair loss of appetite, weight loss, loose rough, dull dull. At the same time, the now toe(referring to) joint swelling, foot dorsal skin edema, ulceration, irritability,restlessness, bite fight, perianal turbidity, the stool is not forming or the thin,breathe bulky external damp invasion of symptoms, drawing model of group spirit is dispirited, addicted to lying, lazy, Shenpi, joint contracture and other symptoms. The symptoms of the treatment group were significantly improved compared with the model group, between the normal group and the model group.Mice weight between the groups comparison results: the time of enrollment,each group of mice were no differences in body recombination; end of the experiment, the control group(30.42 ± 1.72) body weight in mice than the model group(25.64 ± 1.29), statistically, there is a significant difference, P<0.01. The treatment group(27.26 ± 1.17) body weight in mice than the model group(25.64 ± 1.29), statistically, there is a significant difference, P<0.05.Normal group(7.23 ± 0.72) in mice colon segment E. coli count lower than model group(8.49 ± 0.71), statistically, there is a significant difference, P<0.05; treatment group(7.24 ± 0.36) E. coli counts in mice lower than the model group(8.49 ± 0.71), statistically, there is a significant difference, P<0.05. Normal group(6.23 ± 0.94) in mice colon segment lactobacilli counts than the model group(4.66 ± 0.41), statistically, there is a significant difference, P <0.05. Normal group(5.68 ± 0.69) in mice colon segment count of bifidobacteria than the model group(4.99 ± 0.47), statistically, there is a significant difference, P <0.05.Pathological observation of small intestine in mice: the structure of intestinal mucosa was observed under light microscope. The structure of the intestinal mucosa of the mice in normal group was clear, and the shape of the intestinal mucosa epithelial cells was clear. In the model group, the damage of the intestinal mucosa was more serious, the small intestinal villi were broken, the epithelial cells were swollen, and the inflammatory cells infiltrated in the lower part of the intestinal mucosa. The damage of intestinal mucosa in treatment group was less than that in model group, the small intestinal villi were partly broken, the epithelial cells were swollen, and inflammatory cell infiltration was found in the intestinal mucosa.AQP-4, PAR-2, MAPKp38 protein IOD value of the result: the normal group(20.99 ± 2.84), IOD value of the treatment group(17.71 ± 0.77) of the small intestine AQP-4 protein were significantly higher than the model group(14.7 ±1.48), statistically, there is a significant difference, P <0.01 or P <0.05. Normal group(8.62 ± 1.55), IOD value of the treatment group(13.22 ± 1.19) of the small intestine PAR-2 ??protein was significantly lower than the model group(16.76± 1.21), statistically, there is a significant difference, P <0.01. Normal group(8.62± 1.55), IOD value of the treatment group(13.22 ± 1.19) of the small intestine MAPKp38 protein were significantly lower than the model group(16.76 ± 1.21),statistically, there is a significant difference, P <0.01.Serum Cell Factor Results: normal group(166.95 ± 85.56) serum IL-4 were higher than model group(70.51 ± 22.48), statistically, there is a significant difference, P<0.05. Normal group(26.11 ± 14.33) serum IL-6 levels lower than the model group(69.61 ± 33.13), statistically, there is a significant difference, P <0.05.Normal group(73.23 ± 26.17) serum IL-10 were higher than the model group(36.14 ± 22.48), statistically, there is a significant difference, P <0.05. Normal group(51.43 ± 25.35), the treatment group(80.59 ± 34.91) serum IFN-γ content than the model group(138.19 ± 58.81), statistically, there is a significant difference, P <0.05. Expression in mouse intestinal tissue AQP-4 protein in normal group(0.49 ± 0.14) than the model group(0.36 ± 0.13), statistically, there is a significant difference, P <0.05.Western blot analysis Protein: Expression of mouse intestinal tissue PAR-2??protein in normal group(0.18 ± 0.13), treatment group(0.45 ± 0.06)compared with the model group(0.52 ± 0.24), statistically, there is a significant difference, P <0.05. Protein expression in mouse intestinal tissue MAPKp38 the normal group(0.12 ± 0.08), treatment group(0.29 ± 0.15) compared with the model group(0.45 ± 0.12), statistically, there is a significant difference, P <0.05. Expression in mouse intestinal tissue NF-k B protein, the normal group(0.32 ± 0.11), treatment group(0.35 ± 0.07)compared with the model group(0.54 ± 0.11), statistically, there is a significant difference, P <0.05. My D88 protein expression in mouse intestinal tissue, the normal group(0.09 ± 0.08) compared with the model group(0.32 ±0.12), statistically, there is a significant difference, P <0.05. TLR4 protein expression in mouse intestinal tissue, the normal group(0.21 ± 0.10)compared with the model group(0.44 ± 0.16), statistically, there is a significant difference, P <0.05.conclusionsThis study from the Chinese medicine theory, combined with the cold evil,dampness pathogenic characteristics of exogenous cold dampness disease pathogenesis is exogenous cold dampness direct injury of Spleen Yang,exogenous cold dampness further damage to kidney yang, exogenous cold dampness hurt a body Yang, affect the spleen governing transportation functions and Qi of the spleen and stomach machine, body fluid metabolism disorders, spleen dysfunction. Treatment of the external sense cold from the cold dampness legislation.This study successfully established the model of exogenous dampness.Changes in multiple aspects of weight loss, intestinal microflora disorders,enteritis response, disturbance of tissue structure, water metabolism disorders,immune system disorders in model mice with exogenous cold dampness pathogen pathological manifestations.Model group mice intestinal bacteria colony disorder, small intestinal reaction,organization structure disorder changes may be exogenous cold injury of the spleen yang, the influence of the spleen to transport one of the biological basis of function. Abnormal found in mice of model group, water metabolism, may be exogenous cold injury of the spleen yang and the biological basis of block Qi of the spleen and stomach machine, the impact of water metabolism. Model group mice immune system disorders may be exogenous cold injury of Spleen Yang even a body Yang, can affect the immune function of the biological basis. The expression of model group were up-regulated TLR4/ NF-κB signaling pathway, may be one of the molecular mechanisms of exogenous pathogenic dampness.Decline to scattered cold dampness legislation modified kidney a soup to improve the model group, the body weight of mice, intestinal microflora disorders, enteritis reactivity and structure in disorder, water metabolism disorders, immune system disorder status, on / NF-κB signaling pathway inhibition may is scattered Quhan wet lawmaking of modified kidney with an important molecular mechanism of Decoction in treating exopathic dampness. |
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