| Sjogren's Syndrome (SS) in humans is considered a chronic autoimmune disease characterized by diffuse lymphoid cell infiltration into the salivary and lacrimal glands. The consequences of immune cell activation in these exocrine glands are symptoms of dry mouth (xerostomia) and dry eye (keratoconjunctivitis sicca) due to insufficient secretion. SS has no name in traditional Chinese medicine, acceding to its clinic manifestation; it belongs to syndrome caused by dryness-evil. But on diagnosis and therapy, it also adopts the name SS, conducting diagnosis and treatment based on overall analysis of symptoms and signs. As is one of the major damaged organs, and the biopsy is available, salivary gland becomes the focus of SS. By now, the clear pathogenesis of SS is unknown. The lack of appropriate animal models is one of the main causes. In this study, four crude and two pure antigens were used. Through different antigen doses, immune times, sorts and quantities of adjuvant, an mouse model of SS were set up. the sialoadenitis of which was similar to SS in human. On the basis of this model, we investigated the function of T and B cell, researching the relationship between model mice and deficiency of qi and yin. At the same time, we reproduced a rat model, making an inquiry into the blood stasis of model rats. Supplying a powerful tool for the research and therapy of SS through combination of Chinese traditional and Western medicine. 1. Establish mice models of SS1.1 Experimental study on inducing SS-like disease in C57bl/6j mice with crude antigens.Antigen donor animals were four murine strains as follows: C57bl/6j mice, Balb/c mice, SD rats and Lewis rats. The submandibular glands homogenates of above animals were centrifuged at 3000rpm for ISmin.The supernates were mixed with complete Freund's adjuvant (CFA). Then we get the immunogen emulsions with the protein concentration 2 mg/ml (including Bacillus Calmette Guerin (BCG) 3 mg/ml) and injected mice with them, one time every two weeks, total is 6 times and 12 weeks.The results of HE stain showed that antigens from Lewis rats and C57bl/6j mice induced only mild pathologic changes with few lymphocytes infiltrated in submandibular glands (SMG) of C57bl/6j mice, and gentle changes happened in mice with antigens from Balb/c mice and SD rats. The results of double agar diffusion showed that every group presented antibodies to individual antigens, but the groups of-5-Balb/c and SD have higher liters than the other two groups. Nearly no changes wereobserved in group of adjuvant. These changes reflect above experimental conditionscouldn't induce typical pathologic changes in SMG of C57bl/6j mice.1.2 Experimental study on inducing SS-like disease in C57bl/6j mice with pureantigens.Purify the SMG of Lewis rats and C57bl/6j mice by differential centrifugation. Select supernatesS (P5) with two concentrations (l.Omg/ml and 0.5mg/ml). Sensitize C57bl/6j mice on multiple spots with SMG hi CFA plus Bordetella pertussis. Mice were divided into groups from no immunoenhancement to 3 times.The results of HE stain showed that group of P5H3 at 60 days had moderate lymphocytes infiltration in substantial. By day 70, intense lymphocytes infiltration presented in substantial. Generalized infiltration with moderate parenchyma destruction was observed at 80 days. Several, generalized foci of periductal infiltrate with severe parenchyma destruction presented at 90 days. Antibodies increased steady from 60 days to 90 days. The salivary secretion decreased before pathologic changes happened.Above changes reflect we successfully establish a mouse model for SS. 2. The study on pathogenic mechanisms of SS2.1 ImmunopathogenesisInvestigate the function of spleen T and B lymphocyte stimulated by ConA and LPS, and observe the delayed type hypersensitivity (DTK) induced by DNFB. Function of model mice spleen T is decreased and B lymphocyte is increased, DTH is also increased. It reflects the immunoregulation of model mice were disturbance.2.2 Relationship...
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