| Objectives:Take RA (Rheumatoid arthritis) as experimental subject, to quest for the method of making the deficiency of kidney rat model combined with disease and syndrome. For the study of Chinese medicine kidney Arthralgia (RA) and its mechanism and provide experimental tools, and to the disease and syndrome animal model provide further scientific basis.Methods:Take the Deficiency of the kidney rat as combined disease and syndrome model, to estimate the results of the experiment from many aspects such as pathological mechanism and the immune index.Experiment I:In this experiment, choose SPF level forty female Sprague-Dawley (SD) and Wistar rats, twenty female Lewis rats, weight 175±10g, forty male SD and Wistar rats, twenty male Lewis, weight 180±10g. Use cattleⅡtype collagen to make the CIA animal model, adopt the arthritis index, observe briefly and observe under the microscope, to compare the CIA, arthrosis accumulate points and microscope changes.ExperimentⅡ:Make a model according to the mouse species that the experiment I sifts, select and use SPF level 6 weeks SD rats, male and female cutting off testiculus and ovary respectively. Then use cattleⅡtype collagen to make the rat arthritis model after 1 week, 2 weeks,3 weeks,4 weeks. Observing it's arthrosis accumulate points and microscope changes, compare the morbidity and disease severity in different terms of deficiency of the kidney.Results: Experiment I:(1)From the results of statistical analysis, we found that the incidence of different species are significant different; From the pairwise comparison the SD and Wistar rats have notable differences compare with Lewis rats in the incidence of morbidity (P< 0.0001), but the SD and Wistar have no significant difference(P=0.6183); (2) RMANOVA results shown that the AI of different species are significant different (PO.0001). From the pairwise comparison the SD rats have significant difference with Wistar and Lewis rats in AI. Gender have no effect of AI (P=0.737). Immune 28d within the time when compared with other time, there are significant difference. (3)Significance test shown that gender have no significant difference from pathological points but different species of rats have. Pairwise comparison test shown that Lewis have significant difference from SD and Wistar, but SD have no significant difference from Wistar.Experiment II:(1) RMANOVA results shown that Gender have no effect on AI. The rats 4 weeks after emasculation have significant difference with CIA group and the rats 1 week after emasculation. (2)The time of emasculation have significant difference with pathological points but gender have not. (3)Two factor AN OVA shown that gender have significant difference from the level of CORT (P<0.0001). (4)Two factor ANOVA shown that the level of anti-C II have no significant difference from gender but the time of emasculation have. (5) Two factor ANOVA shown that the level of IL-6 and IL-10 have significant difference from gender and the time of emasculation. The level of IL-6 and IL-10 increased when the days of emasculation growth.Conclusions:This study found that:(1)For the induction of CIA, Wistar and SD rats have significant difference from Lewis in the morbidity and AI. But SD rats have significant difference from Wistar in AI. So we recommend the SD rat for CIA animal model. (2)We can choose the 4 weeks emasculation rats for the deficiency of kidney rat model combined with disease and syndrome. (3) Based on the work we had done before, specific production methods are as follows:choose SPF level Sprague-Dawley rats, male or female cutting off testiculus and ovary respectively.4 weeks latter, use cattle II type collagen to make the rat arthritis model. CⅡ(2mg/ml) was dissolved with incomplete Freund's adjuvant at a 1:1 ratio. With a final concentration of lmg/ml. Each rat was injected at the tail root with 200μgCⅡ.7 days latter, Re-immunization with 100μgCⅡeach again. |
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