| Objective: To objectively evaluate the regulation of spleen invigorating and kidney moisting therapy combined with Western medicine upon the epigenetic microRNA in patients with systemic lupus erythematosus(SLE)at acute period,and to explore the possible molecular mechanism of two Chinese Medicine therapies and their significance for SLE treatment through analyzing the changes in serum MicroRNA expression profile before and after treatment.Methods: 1.SLE patients subjects,who were admitted to Departments of Dermatology and Rheumatism Immunity,Hospital Affiliated to Chengdu University of TCM from September 2014 to December 2015 and met the inclusion and exclusion criteria,were randomly divided into the treatment group and the control group,which were treated respectively with Zhibaidihuang Pill combined with Sijunzi Decoction+Prednisone Tablet+Hydroxychloroquine Sulphate vs.Zhibaidihuang Pill+Prednisone Tablet+Hydroxychloroquine Sulphate for 9 months;the efficacy indicators included 1)SLEDAI,SLE syndrome grading scale,and 2)complement,ds-DNA,24-hour urinary protein;the safety indicators included blood routine,liver function and renal function.2.Five healthy volunteers were selected as healthy controls.3.5ml of fasting venous blood was collected from the healthy control group,the treatment group and the control group before and after treatment to extract the total RNA for microRNA gene chip detection;biological analysis and target gene prediction were conducted for the differential microRNAs.4.Select TargetScan,miRTarBase to predict the target gene.5.SPSS 17.0 software was used for the statistical analysis,where the inter-group counting data were compared using chi-square test(Fisher’s exact test would be used if n<40)and the continuous variables were represented by mean ± standard deviation.Kolmogorov-Smirnov normality test was performed in all samples.The data inconsistent with normal distribution were submitted for rank sum test;for those consistent with normal distribution,independent samples t test was conducted for inter-group comparison while paired-sample t test for intra-group comparison.The significant level was defined as α=0.05.Results: 1.After treatment,SLEDAI scores in the treatment group were significantly reduced(P<0.05),so were the scores of SLE syndrome grading scale in the treatment and control groups(P<0.05).2.Complement,ds-DNA and 24-hour urinary protein were improved in the treatment and control groups after treatment,but the differences were not statistically significant(P> 0.05).3.The levels of blood routine,liver function and renal function in the treatment and control groups were within the normal range both before and after treatment,suggesting no statistically significant differences.4.Subject to the criteria of disease therapeutic effect,the treatment group that involved 5 cases reported 3 cases of marked effectiveness and 2 cases of effectiveness,indicating a total effective rate of 100%;the control group that involved 3 cases reported 1 case of marked effectiveness and 2 cases of effectiveness,indicating a total effective rate of 100%.The total effective rate of these two groups was identical.5.Compared with the healthy volunteers(G1)before treatment,58 microRNAs with significant differences were obtained in the treatment group(G2),including 10 upregulated ones and 48 downregulated ones;compared with the treatment group before treatment(G2)after 9 months of treatment,1 miRNA(miR-22-3p)was upregulated in those 58 microRNAs while 3 ones(miR-1246,miR-4644 and miR-6739-5p)were downregulated.The test suggested no microRNAs with treatment-related significant differences in the control group.6.The target gene prediction for differential microRNAs and the enrichment analysis on GO and protein signaling pathways(KEGG)concluded 4 signaling pathways closely associated with SLE,namely,ubiquitin-proteasome system,TGF-β signaling pathway,Notch signaling pathway and p53 signaling pathway;the related biological processes participating in regulating material metabolism included mucopolysaccharide biosynthesis,fatty acid elongation,unsaturated fatty acid biosynthesis,mineral absorption,and calcium absorption regulated by endocrine and other factors.7.The core valuable microRNAs are miR-4516、miR-486-5p、miR-642a-3p、6749-5p、miR-4281、miR-6088、miR-6869-5p、miR-6087、miR-6090、miR-3162-3p、miR-6800-5p、miR-4763-3p、miR-3960、miR-451a、miR-6821-5p,which come out after time series analysis of Group1-Group2-Group3-Group4.Conclusion: 1.Incorporating the conventional Chinese medicine for yin nourishing and heat clearing combined with Western medicine,the spleen invigorating and kidney moisting therapy combined with Westen medicine is safe,effective and free of adverse reactions in the treatment of SLE at acute period and exerts greater effect in improving SLEDAI scores than the conventional therapy.2.One molecular mechanism of the spleen invigorating and kidney moisting therapy combined with Westen medicine in the SLE treatment may function through regulating certain apoptosis-or material metabolism-related signaling pathways via affecting the expression of related microRNAs,thus realizing immune regulation and improvement in physical fitness.3.“Preventive Treatment of Disease” as the guiding ideology of the spleen invigorating and kidney moisting therapy combined with Westen medicine in the treatment of SLE acute phase important. |
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