Differential Expression And Clinical Significance Of Mirna In Mononuclear Cells Between Crohn’s Disease And Intestinal Tuberculosis

Inflammatory bowel disease (IBD) includes ulcerative colitis (UC) andCrohn’s disease (CD), which incidence increases every year in China. However,it’s still difficult to make the accurate diagnosis and treatment. Because theincidence of intestinal tuberculosis (ITB) is high in China, which takes similarsymptoms, colonoscopic findings and pathological changes as IBD. Thoughsome ways have come out at home and abroad, it’s still difficult make thedifferential diagnosis.Tuberculosis T-SPOT (T-SPOT.TB) is the most valuable detection methodfor active tuberculosis infection. Through stimulating circulatory blood bytuberculosis specific antigen CFP-10and ESAT-6, it sensitized mononuclearcells to release interferon-r. And then we can measure the latent or currenttuberculosis infection through immunohistochemistry for precipitation spots.The sensitivity and specify of T-SPOT.TB comes to over95%[1]， which isgetting more and more widely used in clinic. Though a good method for tuberculosis detection, it has been used for differential diagnosis between CDand ITB. This study primarily studied on the differential diagnosis value ofT-SPOT.TB in CD and ITB, which showed good result in active CD and ITB.However, it’s still difficult to make accurate differentially diagnosis on moderateor remission CD and ITB.It has been reported the diagnosis value of microRNA (miRNA) as themarkers for CD and UC. Meanwhile, different miRNA can be markers fordifferent tumor. It has also specific miRNA expression in IBD tissue. But it’sunclear if different miRNA exist in the un-stimulated or stimulated mononuclearcells of CD and ITB. After stimulating the mononuclear cells with tuberculosisspecific antigen CFP-10and ESAT-6, miRNA array was used to analyze thedifferential expressed miRNA in CD and ITB patients to identify the valuablemiRNA in differential diagnosis for CD and ITB.Aim1. To study on the value of T-SPOT.TB in differential diagnosis of CD andITB;2. To compare the different miRNA in CD and ITB patients mononuclear cellsafter stimulating with tuberculosis specific antigen CFP-10and ESAT-6;3. To verify the different miRNA in CD and ITB patients with Real-time PCR.Methods1. To collect the circulatory blood from126CD, ITB and non-ITB patients forT-SPOT.TB detection.2. To compare the differential expression of miRNA through3healthyvolunteer,3active CD and ITB patients mononuclear cells after stimulatingwith tuberculosis specific antigen CFP-10and ESAT-6. 3. To analyze the statically differential expressed miRNA by bioinformatics.4. To verify the differential expressed miRNA in5CD and ITB patients’ coloncontrast normal colon through Real-time PCR.Results1.15patients were diagnosised as Crohn’s disease(11.9%,15/126),14patientswere intestinal tuberculosis (11.1%,14/126) and40patients were non-ITB(31.7%,40/126).The positive rate of T-SPOT.TB in Crohn’s disease,intestinaltuberculosis, non-ITB and other disease was6.7%(1/15),85.7%(12/14),70%(28/40) and0%(0/57),respectively.The difference between the groupswas statisically signifficant (P=0.001).There was statistically significantdifference of T-SPOT.TB positive rate between Crohn’s disease andintestinal tuberculosis (χ2=70.58，P=0.001).The sensitivity and specificity ofT-SPOT.TB in Crohn’s disease detection was93.3%(14/15)和87.5%(14/16),in intestinal tuberculosis was85.7%(12/14) and93.3%(14/15). Thenegatively predictive value of Crohn’s disease was higher [87.5%(14/16)]than that of intestinal tuberculosis [12.5%(2/16)].2. To compare the differential expression of miRNA between CD and ITB, itshowed12upregulated and10downregulated miRNA in ITB. To comparethe differential expression of miRNA between CD and ITB patients’mononuclear cells after stimulating with tuberculosis specific antigenCFP-10and ESAT-6, it showed9upregulated and1downregulated in ITB.We found that the relationship between the regulation network analysis oftarget genes and difference miRNAs,7miRNAs upregulated anddownregulated respectively.After comprethensive analysis, miR-125b,miR-31, miR-19b, miR-939and miR-144showed most significant difference finally.3. It has been verified that miR-31, miR-19b, miR-939and miR-144showedno significant difference between CD and ITB patients by Real-time PCR,except for miR-125b. The mean expression value of miR-125b is1.001involunteer, which is significantly upregulated in ITB patients (1.161) anddownregulated in CD patients (0.679)(P <0.01).Conclusion1. T-SPOT.TB is an effective differential diagnosis method for active CD andITB;2. The expression of miR-125b shows significant difference in CD and ITBpatients, which is high in ITB and low in CD. miR-125might be a valuablemarker for the differential diagnosis of CD from ITB.