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Objective:To investigate the expression level of plasma miR-320c in patients with osteoarthritis(OA), and explore the clinical significance and the role in pathogenesis of OA.Methods:The clinical data and peripheral blood of 30 patients with OA, 30 patients with connective tissue diseases and 30 healthy control individuals were collected.The levels of plasma miR-320c were detected byfluorescentquantitative reverse transcription PCR(qRT-PCR). Correlation analysis was used to explore the correlation of plasma miR-320c level with knee X-ray data and VAS pain score in OA patients.Finally the miR-320c mimic, the miR-320c inhibitor, and the control material were transfected to the chondrocyte HC-a.The proliferative capacity of HC-a chondrocytes was examined at different time points as determined by the CCK-8 assay.Results:The expression level of plasma miR-320c was significantly higher in OA group(3.26±0.55)than that in the connective tissue diseases group(1.62±0.50)and in healthy control group(1.21±0.66)( F=107.66, P<0.001). Plasma miR-320c expression was positively correlated with radiographic grade( r=0.830, P<0.001), and had no correlation with VAS pain score in OA group( P>0.05). Through repeated measurement variance analysis, the time effect, the group effect and the interaction effect between group and time showed statistically significant differences in chondrocyte proliferation between NC mimic group and miR-320c mimic group( Ftime=5256.767, Fgroup=1947.436, Ftime×group=114.314, all P<0.001). The level of proliferation was significantly reduced.Apoptosis rate of chondrocytes was significantly increased in the group transfected with miR-320c( t=7.85, P<0.01). Conclusions:The expression level of plasma miR-320c is significantly higher in osteoarthritis patients and associated with knee radiographic severity grade.Furthermore, over-expression of miR-320c could suppress the proliferation of chondrocytes.Plasma miR-320c might be potential bio-marker for osteoarthritis knee severity assessment, and involves in regulating chondrocyte growth in the pathogenesis of osteoarthritis.
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Objective:To explore the characteristics of plasma microRNA (miRNA) profiles and bioinformatics in patients with osteoarthritis(OA) in order to search for diseases related biomarkers.Methods:Blood samples from 20 cases of OA patients and 15 cases of normal control (NC) were collected to extracted total RNA in plasma. The plasma miRNA expression profile was tested by using Agilent Human miRNA array. Target gene analysis and clustering analysis were performed on differentially expressed microRNAs. Three differentially expressed miRNAs (miR-134-5p, miR-320c and miR-940) were detected by real-time quantitative polymerase chain reaction (RT-qPCR) for further confirmation of microarray data. The differences were tested using t test analysis. Results:① MiRNA microarray showed that compared with NC, there were 74 differential expression genes in plasma of patients in the OA group (FC≥2, P≤0.01), among which 45 were up-regulated and 29 were down-regulated. ② A total of 2 731 potential target genes were predicted in three database, and involved in 462 Kyoto Encyclopedia of Genes and Genomes (KEEG) pathways. Target gene ontology (GO) functional clustering found that the main functions of miRNAs were intercellular adhesion, collagen synthesis, intracellular signal transduction, etc. The main KEGG pathways of miRNAs include mitogen-activated protein kinase (MAPK) signaling pathway, cyclic adenosine monophosphate (cAMP) signaling pathway, osteoclast differentiation signaling pathway, etc. ③ The expression level of miR-20a-5p and miR-320c in OA group were significantly higher than that in controls ( t=-6.142, P<0.05; t=-3.854, P<0.05), while miR-940 was significantly lower than that of controls ( t=2.767, P<0.05) . The trend was consistent with the microarray data. The receiver operating characteristic curve (ROC) curve analyses showed that they were useful biomarkers for differentiating patients with OA from controls. Conclusion:The study shows that plasma in OA patients has a specific miRNAs expression, and miRNAs play an important role in the pathogenesis of OA.
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Objective To explore the effect of different concentrations of iguratimod (IGU) on mouse model of bleomycin-induced pulmonary fibrosis.Methods A total of 108 female C57BL/6 mice were randomly divided into the control group,the model group,the low dose IGU group,the moderate dose IGU group,high dose with group and the methylprednisolone (MP) group (n=18 in each group).The mice in the control group were injected with 0.2 ml normal saline endotracheally,and others were injected with 0.2 ml bleomycin (5 mg/kg) from endotracheally to induce pulmonary fibrosis model.The next day,the mice in both control group and the model group were fed with 0.2 ml normal saline every day;The mice in the IGU groups and methylprednisolone group were fed with 0.2 ml iguratimod liquid the IGU (10,30,90 mg/kg) and 0.2 ml methylprednisolone (10 mg/kg) every dayrespectively.Finally the mice were sacrificed at day 7,day 14,day 28 respectively,and the lung tissue was examined by HE staining and Masson staining to evaluate the degree of alveolitis and fibrosis.Repeated measurement of variance analysis was used to compare the differences for time and group,and multi-factor analysis of variance LSD test was used for the comparison between groups.Results ① The body mass of mice in bleomycin-induced groups were decreased compared to the control group.② The pathological alveolitis scores in the high dose IGU group and methylprednisolone group were significantly decreased compared to those of the model IGU group at day 7 and day 14 (P＜0.05),and the pathological fibrosis scores were decreased dramatically compared with the model group at day 14 and day 28 (P＜0.05).Conclusion High concentration of IGU and methylprednisolone can reduce and inhibit lung inflammation and fibrosis of bleomycin-induced pulmonary fibrosis in mice.
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Objective To investigate the expression of interleukin-35 (IL-35) in serum of mice with pulmonary interstitial fibrosis.Methods Thirty-six C57BL/6 mice were divided into three groups (twelve mice in each group):control group,model group of mice with bleomycin-induced pulmonary fibrosis,glucocorticoid treatment group of mice with bleomycin-induced pulmonary fibrosis.The mice were sacrificed at day 7,day 14 and day 28 respectively,and the serum concentration of IL-35 was assayed.Statistical product and service solutions (SPSS) 17.0 statistical software was used for single factor analysis of variance and LSD-t comparison and Pearson correlation analysis was used for the comparison between each two groups.Results The serum IL-35 concentrations between groups and within groups at the time of day 7,day 14 and day 28 were compared respectively.The serum IL-35 concentration of the model group was significantly lower than the control group and the glucocorticoid treatment group at the time of day 7 (F=24.56,P＜0.05).The serum IL-35 concentration of glucocorticoid treatment group was significantly lower than the control group at the time of day 14 (F=8.85,P＜0.05),and which of glucocorticoid treatment group was significantly lower than the control group and the model group at the time of day 28 (F=36.64,P＜0.05).There was no significant difference between days 28 and day 7 within control group (t=1.03,P＞0.05).The serum IL-35 concentration of the model group at the time of day 28 was significantly higher than those of day 7 [(9.36±0.95) ng/ml vs (6.51±0.58) ng/ml,t=5.14,P＜0.05],and which of glucocorticoid treatment group was significantly lower than those of day 7 [(5.27±1.01) ng/ml vs (9.42±0.84) ng/ml,t=6.32,P＜0.05].From day 7 to day 28 the serum IL-35 concentration change of the model group and glucocorticoid treatment group showed significantly negative correlation (r =-0.743,P＜0.05).Conclusion Serum IL-35 concentration has shown an trend of increase during bleomycin-induced pulmonary fibrosis with mice model,and early glucocorticoid treatment can decrease the serum IL-35 in the model mice.
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Objective To investigate the inhibitory effect of methylprednisolone (MP) on the development of coronary arteritis in a murine model of Kawasaki disease (KD) induced with a candida albicaus watersoluble fraction (CAWS).Methods Forty-five C57BIL/6mice were evenly divided into three groups (the control group,the CAWS group and the MP group).Mice in the CAWS group were intraperitoneally injected phosphate buffer saline (PBS) for 5 days.MP or PBS was administered to the different group.The animals were sacrificed at day 3,day 10 and day 28,and the status of vasculitis in the coronary arteries and the aortic root was investigated histologically.One-way analysis of variance (ANOVA) was used to compare the differences among three groups,and t-test for two independent groups.Results The mice in CAWS group and MP group,which induced by CAWS,showed that the body weight and heart weight decreased significantly,and the spleen weight was increased at day 10 and day 28 (P＜0.05).Vasculitis was induced in the mice of those two groups,and the severity score was the highest at day 10 (12.7±0.5).In addition,the severity of the inflammation of the aortic root and the coronary arteries were reduced in MP group (t=6.35,5.55,2.8,P＜0.05).Elastic fiber staining showed that the layers of vascular walls were in disorder and elastic fibers were broken in the CAWS group.However,there was no disruption or breakage in the MP group.Conclusion MP can suppress the progression of coronary arteritis in this CAWS-induced murine vasculitis model,which indicates the efficacy of MP in KD patients with coronary artery lesions.
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Objective To prepare RGD and TAT co-modified paclitaxel loaded liposome(RGD/TAT-LP-PTX)for A 549 cells targeting.Method The co-modified liposome was prepared by film-ultrasonic method. The appearance,particle size,Zeta potential were evaluated. The cellular uptake by A 549 cells in vitro was used to evaluate the targeting efficiency. The anti-proliferation efficiency of RGD/TAT-LP-PTX was evaluated by MTT assay. Results The particle diameter of the co-modified liposome was (118.5±11.4) nm with the Zeta potential of (21.58±2.42 )mV. The entrapment efficiency of PTX was 86.5%. The result demonstrated that the co-modified liposome uptaken by A 549 were 2.1, 2.8 times higher than that of TAT-LP and RGD-LP, respectively. The RGD/TAT-LP-PTX shows the highest anti-proliferation efficiency. Conclusion The co-modified liposome might serve as a promising tumor delivery system of antitumor drugs.