Clinic Diagnostic Significances And Values Of Circulating MiR-189 In System Lupus Erythematosus

System lupus erythematosus(SLE) is an autoimmune disease that predominantly affects women of childbearing age. Cases about the old and the child are reported too. However till today, it is not clear about the pathogeny and the mechanism of SLE. Accroding to research reportes related, the propable pathogeny of SLE includes heredity, circumstance, medicine and disorder of autoimmune system. The main mechanism results in the change of the ratio between T and B cells and the over-actived in their function. It causes the immunoreaction between autoantibody and auto reactive lymphocytes or antigens which leads to lesion of auto tissues and disorder of normal function and then arises the development of SLE.Because SLE is related to several tissues and several organs, there are manifold clinical manifestation in patients with SLE. It may be character as a system or a prominent characteristic of clinical manifestation and then is misdiagnosed as other diseases. Because of having missed the best treatment, tissues and organs are injured which can not be reversed after treatment.Considered the particularity of SLE in pathological damage and clinical shows, the standards of clinical diagnosis are based mainly on clinical performance and clinical laboratory diagnosis which pay the key role in disease diagnosis and evaluation of therapeutic efficiency. The rapid detection technology of high sensitivity and high flux for laboratory screening is benefit for the project developed and expanded, for the disease early detection and early diagnosis and early treatment. At present, a screening program of SLE and other autoimmune diseases is antibody's spectrum which includes anti-dsDNA, anti-ssDNA, anti-nuclear antibodies and anti-extractable nuclear antigens(ENA). Anti-dsDNA antibody is a most important biomark for SLE. The complex of dsDNA and its antibody can lead to damage of subcutaneous tissues, the kidneys and other organs. There has a correlation of antibody titre and state of an illness. In addition, another biomark for SLE is anti-Sm antibody which is supposed to be involved in the pathological process. The specificity of two antibodies is high, but the sensitivity is relatively low. So a few of researchers are applying themselves to explore a more unique and more reliable molecular biomark as the supplement or replacer for the antibodies for diagnostic in SLE.MicroRNA(miRNA) is a 19-23nt long non-coding RNA sequence that is involved in various biological processes, including cell proliferation, differeation, apoptosis, stress response and tumorigenesis. MiRNA is complementary to 3' UTR sequence motifs that mediate negative post-transcriptional regulation which affects cell development and diseases progression through the up or down regulation expression. A number of miRNAs are specific expression in cells or tissues, so the changes in the expression levels of miRNAs may be related to certain diseases patterns.Recently, circulating miRNA has been reported as potential biomarker for the cancer, myocardial infarction, hepatic injury et al. miRNA is more stable in plasma or serum which is different from other RNA molecules. Its expression is ofter stable in normal state. It is not been degraded by enzymes in the blood that would degrade regular RNA. MiRNA remain stable even after incubation at room temperature and after many freeze/thaw cycles. Base on requirement of clinic and the special character of miRNA, whether miRNA can be used as a biomark for complementary diagnosis for SLE or not. Therefore, this reasearch is to test relative expression of microRNA189(miR-189) in circulating blood and explored the values in clinic diagnosis and the meaning for SLE.Purpose:To explore whether there has the difference in SLE group and control groups and the potential diagnostic value for SLE of miR-189. Whether miR-189 can make up for the deficiency of autoantibody repertoire for diagnosis and therapeutic evaluations with SLE.Methods:1. Autoimmune antibody detectionAutoantibody repertoire of Specimen which are collected are tested. The main method adopted is immunization. Anti-dsDNA tests and anti-ssDNA tests are adopted by a measure of ELISA indirect method. Anti-ENA tests are adopted by immune blotting. ANA tests are adopted by immunization fluorescence detection method. All these are finished by operating instruction.2. Circulating miR-189 relative quantificationSample serums of SLE patients(n=50) included stable phrase (n=13) and active phrase(n=37), normal control person(n=24) who come form healthy people physical examined and disease control patients(n=25) included behcet disease(BD)(n=2),system sclerosis(SSc)(n=1),sjogren's disease(SS)(n=1),polymyositis/ dermatomyositis(PM/DM)(n=4),rheumatoid arthritis(RA)(n=7),ankylosing spondylitis(AS)(n=5),mixed connective tissue disorder(MCTD)(n=5) are Collected. There has no significance of age and gender in three groups. Total RNAs are extracted from serums applied RAN special extraction kit form ABI company and reversed to cDNA. TaqMan probe was applied to Real-time PCR detection. At last, relative quantification of miR-189 is compared among three groups.Resulst:1.Comparison of antibody's positive percentage in three groupsPostive percentage about ANA, anti-dsDNA, anti-Sm, anti-Rnp and AnuA is calculated among three groups. They are 98%, 42%, 28%, 18% , 24% separate in SLE group and 19%, 4%, 0%, 2%, 1% separate in diseases control. the specificity of these biomark is 24%,84%,100%,92%, 96% separate in SLE group. The conclusion is when the specificity is high while the sensitivity is low about anti-dsDNA, anti-Sm,anti-Rnp, AnuA or the sensitivity is high while the specificity is low about ANA.2. Comparison of relative quantification(RQ) of miR-189 in three groupsRQ of miR-189. SLE patient group (26.8713±23.1291) is highest Among three groups(disease control group, 5.2281±4.4251, normal control group, 3.9908±4.0281). There has obvious significance (p=0.000). Two control groups have no significance(p>0.05). stable phrase(34.8932±27.4931) is higher than active phrase(24.0528±21.0898), but there has no significance(p=0.2885).3. RQ of miR-189 and correlationThere has no significant correlation between the relative expression of miR-189 with SLEDAI scores(r=0.009, p=0.968), the dose of prednisone(r=-0.2376, p=0.1831). and anti-dsDNA(r=0.2607, p=0.2484) in SLE patients.4. Diagnositic values of RQ with miR-189 for SLECompared to normal control, diagnositic values of RQ with miR-189 for SLE. Area under the curve(AUC) is 0.888 and standard error is 0.036. there has diagnostic values of RQ with miR-189(p=0.000). The large of RQ is higher, the probality of SLE is more. Confidence interval(CI) is from 0.817 to 0.960, 0.5 is not included. So there has significance of diagnosis. Compared to normal control, diagnositic values of RQ for other immune diseases. AUC is 0. 583 and stand error is 0.083. there has no diagnostic values. Compared between SLE group and disease control group, diagnositic values of RQ with miR-189 for SLE. AUC is 0. 847 and stand error is 0.043, there has diagnostic values of RQ with miR-189(p=0.000) which distinguish other immune disease.Conclusion:1. Auto antibodies spectrum are mainly adopted to help diagnose autoimmune diseases including SEL for clinical. From the results, the unity of specificity and sensitivity is difficult. Misdiagnosis is easily occured when sensitivity is high and specificity is low. Misseddiagnosis is easily hanppened when specificit is high and sensitivity is low. There is obvious dificiency.2. From ROC curve analysis, circulating miR-189 can be used as a potential diagnostic biomark for SLE which can help or supplement the laboratary tests.RQs of stable phrase and active phrase are all high compare to control, they illuminate the morbid state and no diagnostic values for severity of disease. they have no change when the severity of disease is changed. MiR-189 can be used as a diagnostic marker for early diagnosis and make patients receive treatement in early stage.