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Purpose and significance of SLE (systemic lupus erythematosus, SLE) is an autoimmune mediated autoimmune inflammation for the outstanding performance in diffuse connective tissue disease, is considered to be the prototype of autoimmune diseases, in order to appear in the serum anti- nuclear antibodies (antinuclear antibody, ANA) as the representative of a variety of autoantibodies and multi-organ, multi-system involvement as the main clinical features. SLE is widely distributed in the world, the world prevalence rate of about 17 to 48/100 000, China's textile workers in Shanghai conducted a large series of one-off surveys show SLE prevalence rate of 70/10 people, women is as high as 113/100, 000, slightly higher than Western countries. SLE occurs in young women, more common in 15 to 45 years old, male to female ratio of 1:7 ~ 9. SLE involving the kidney is the renal damage caused by lupus nephritis (lupus nephritis, LN). LN SLE is the most common and serious damage to internal organs, according to the clinical manifestations of patients with more than 70% of renal involvement, if according to renal tissue biopsy, almost all SLE patients had kidney damage, the severity of their kidney disease is a direct impact on SLE an important prognostic factor. LN complex and diverse clinical manifestations, of varying severity, can be manifested as proteinuria, hematuria and urinary tube, or with hypertension, edema and other clinical kidney damage and renal dysfunction in experimental basis, clinical diagnosis and treatment difficult, resulting in the gradual progress of the disease, Some patients may eventually progress to ESRD (end-stage renal disease, ESRD), one of the main causes of death in SLE. LN etiology and pathogenesis is not very clear at present that is the basis of genetic susceptibility by environmental factors (including infection, sunlight, climate, food, drugs, etc.), endocrine (primarily estrogen levels, which may female incidence was significantly higher than the male one of the reasons) induced, caused by the body's immune function disorders, T, B lymphocytes abnormal activation of cytokine network imbalance, complement activation and apoptosis, leading to ANA, anti-dsDNA antibody, anti-Sm antibodies and other pathological autoantibodies and immune complex formation. These autoantibodies against nuclear antigens main component, especially for nuclear DNA antibodies, formation of antigen-antibody complexes deposited in the target organ vessel wall, triggering a series of immune response, leading to multiple organ damage the body. Itself in the glomerular deposition of immune complexes after complement activation, resulting in inflammatory cell infiltration, activation of coagulation factors and a variety of inflammatory mediator release, resulting in kidney damage. Modern medicine with corticosteroids, cytotoxic drugs, a variety of new methods, such as biological agents, plasmapheresis, stem cell transplantation, the impact of high-dose immunoglobulin therapy and other treatment LN, achieved a certain effect. But the recurrence rate of these drugs, side effects, expensive, many patients are often difficult to adhere to long-term treatment, and because of SLE complexity and heterogeneity, simply one part of the therapeutic intervention strategies are often difficult to obtain good results. The medicine is one of the major Chinese medical treasure trove of resources, clinical practice showed that Chinese medicine has a multi-site, multi-link and multi-target overall regulatory function and improve efficacy, improve prognosis and reduce the recurrence of the disease to maintain long-term stability, helping to stop the use of hormones to maintain or reduce the amount of hormones and side effects, and so has a good effect on multiple genes, complex pathogenesis, disease varied LN patient has very important significance. TGP (1otal glucosides of paeony, TGP) Product name Pavlin, as the first anti-inflammatory immunomodulatory drugs are officially approved for production listed for rheumatoid arthritis (rheumatoid arthritis, RA) clinical treatment. TGP is after years of research scholars from the traditional Chinese medicine Ranunculaceae drying plant peony root extract the active ingredients, mainly for a group of glycosides substances, including Paeoniflorin, hydroxyl Paeoniflorin, peony flower glycosides, albiflorin , benzoyl Paeoniflorin etc., collectively known as TGP, which Paeoniflorin total amount of more than 90% glycosides, the main active ingredient is white peony. Pharmacological and clinical studies have found that it has good anti-inflammatory, analgesic and immunomodulatory effects, and adverse reactions less well tolerated by patients. In some autoimmune diseases, early and remission, should not be applied potent immunosuppressant or hormone-treated patients, TGP is satisfied with the choice. Has been gradually applied to systemic lupus erythematosus, ankylosing spondylitis, Sjogren's syndrome and other autoimmune diseases, in the treatment of autoimmune diseases shows unique application value and good prospects, but the clinical reports for LN rare, experimental studies have not been reported. MRL / lpr mice, by the United States Jackson Laboratory Murphy and Roths successful cultivation in 1978, is currently the most commonly studied SLE and LN one spontaneous animal model. The mice with cells containing spontaneous programmed cell death-related Fas gene recessive mutations appear lymphoproliferative gene (1ymphoproliferation, lpr), autoreactive T cells that may escape thymic negative selection, a large release into the peripheral organs, resulting in T cell hyperplasia and accelerated autoimmune reaction, spleen and lymph node enlargement occurs, a variety of autoantibodies, high agammaglobulinemia and related immune complex glomerulonephritis, similar to human lupus nephritis. This study intends to adopt MRL / lpr mice as experimental subjects, observation of TGP on MRL / lpr mice with lupus nephritis urine protein, serum anti-dsDNA antibodies, ANA, renal histopathology, spleen and thymus index and the general condition of the implications for TGP further explore the mechanism of immune regulation and clinical treatment LN provide important theoretical and experimental basis. Methods and content 1. Experimental groups: MRL / Lpr mice by quarantine, adaptive feeding one week after the detection of MRL / lpr mouse anti-dsDNA antibodies, ANA levels, urine protein, anti-dsDNA antibodies, ANA-positive and proteinuria ≥ 100mg-dl-1 pathogenesis prompted to establish the model. Digital table grouping were randomly divided into model group and TGP group (TGP), n = 10. Accordance with the \2. Detection methods: (1) mice were observed before and after treatment mental state, hair, food, activities and other general changes in circumstances. (2) before treatment, after a fixed time 15,30 d morning Weigh weight, tail lift reflection method using urine specimens stored at -80 ℃, Coomassie brilliant blue concentrated urine protein content. (3) before treatment, the morning after a fixed time 15d tail vein blood after treatment 30d eyeball blood, centrifuged and the serum stored at -80 ℃, double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) centralized testing Serum IgG anti-dsDNA antibodies, ANA levels. (4) 30d after treatment mice were sacrificed, spleen and thymus, pat it dry with a paper surface bloodstained called wet weight, respectively, were calculated spleen and thymus index. Take kidneys, are fixed, paraffin-embedded, hematoxylin side of the kidney - eosin (HE) staining, pathological changes observed under light microscope. Other side of the kidneys immediately set -70 ℃ freezer kept aside. 3 Statistical analysis: SPSS13.0 statistical software analysis, measurement data were expressed as mean ± standard deviation (X ± S) said that, spleen, thymus index comparison of the quality and homogeneity of variance test, using two independent samples t-test . Weight, urine protein autoantibodies comparison of the spherically symmetric hypothesis testing, using repeated measures analysis of variance design, multiple comparisons using the least significant difference method (LSD) test. P lt; 0.05 as statistically significant difference. Results 1. General situation: two groups of mice before treatment Maofa Run Ze smooth, feeding, activity sensitive. Over time, the gradual emergence of model mice scattered sparse hair, dull, back of the neck fur of individual mice was significantly off; mental fatigue, hi lying curled, activity decreased activity more slowly, feeding less than before. TGP mice lighter hair less shiny, freedom of movement, feeding change is not obvious; end of the experiment were no deaths (2) Weight: TGP body weight of mice after administration of 30d (23.43 ± 3.60) than after administration of 15d (22.49 ± 4.35), the difference was statistically significant (P lt; 0.05). Model body weight of mice after administration of 30d (21.59 ± 2.75) than after administration of 15d (22.83 ± 3.25) reduced, the difference was statistically significant (P lt; 0.05). 3 spleen, thymus weight and index: After 30d, TGP group of mice spleen weight and its index (89.20 ± 47.00), (4.02 ± 2.38) compared with model group (132.70 ± 41.40), (6.31 ± 2.34), difference was statistically significant (P lt; 0.05). Two groups of mice thymus weight and its index, the difference was not statistically significant (P gt; 0.05). 4 urinary protein content: (a) after treatment 15,30 d, TGP mice urine protein content (100.40 ± 23.89), (93.79 ± 22.03) compared with the model group (122.63 ± 14.20), (127.88 ± 14.03), the difference There was statistically significant (P lt; 0.05, P lt; 0.01); (2) TGP mice urine protein levels after treatment 15 (100.40 ± 23.89), 30d (93.79 ± 22.03) than before treatment (115.61 ± 13.09) lower The differences were statistically significant (P lt; 0.05); (3) model mice urine protein levels before and after treatment showed no significant difference (P gt; 0.05). 5.IgG anti-dsDNA antibody levels: (a) after treatment 30d, TGP mice IgG anti-dsDNA antibody levels (847.80 ± 577.68) compared with the model group (1893.17 ± 610.33), the difference was statistically significant (P lt; 0.01); (2) TGP anti-dsDNA antibody levels in mice after administration of 30d (847.80 ± 577.68) than before treatment (1260.82 ± 942.70), after administration of 15d (1278.18 ± 825.16), the difference was statistically significant (P lt ; 0.05). (3) model mice IgG anti-dsDNA antibody levels after treatment, 15 (1783.27 ± 829.70), 30d (1893.17 ± 610.33) than before treatment (1273.17 ± 767.13) gradually increased, the difference was statistically significant (P lt; 0.05, P lt; 0.01). 6.ANA levels: (a) after treatment 30d, TGP mice ANA antibody levels (158.76 ± 66.86) compared with the model group (419.65 ± 162.04), the difference was statistically significant (P lt; 0.01); (2) TGP ANA antibody levels in mice after administration of 15 (241.72 ± 105.43), 30d (158.76 ± 66.86) than before treatment (385.18 ± 135.91), the difference was statistically significant (P lt; 0.05, P lt; 0.01); (3 ) model mice ANA antibody levels before and after treatment showed no significant difference (P gt; 0.05). 7 Renal Pathology: model mice mesangial cells and endothelial cells, accompanied by inflammatory cell infiltration, thickening of the basement membrane is not obvious, some glomerular volume increased significantly; tubular epithelial cell swelling and vacuolization degeneration seen in the lumen tube. TGP mice compared with the model group were glomeruli and renal interstitial infiltration of inflammatory cells was significantly reduced, tubular mild turbidity inflation. Conclusion 1.TGP significantly improved MRL / lpr mice with lupus nephritis and general condition of the mice weight gain, enlarged spleen alleviate that TGP can improve the MRL / lpr mice with lupus nephritis quality of life, and has a protective immune organs effect. 2.TGP can reduce serum ANA, anti-dsDNA antibody levels, suggesting that TGP has a regulatory role on the immune system. TGP immunoregulatory network possible through a pathway to inhibit the production of autoantibodies, reduce their level of antibody induced immune response, and thus the treatment LN. 3.TGP can reduce the MRL / lpr mice with lupus nephritis mesangial cells and endothelial cells, reduce inflammation glomerular and tubular injury and improve renal pathological changes, reduce urinary protein content. TGP with slow disease progression showed protect kidney function. 4.TGP on MRL / lpr mice with lupus nephritis have a certain effect, as the underlying mechanism of its functioning, future studies will be discussed.
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