ABSTRACT
Objective To investigate the effect of microRNA(miR)-30d-5p on osteogenic differentiation and apoptosis of bone marrow stromal cells and its mechanism. Methods Bone marrow stromal cells were divided into miR-30d-5p overexpression negative control group, miR-30d-5p overexpression group, miR-30d-5p inhibition negative control group and miR-30d-5p inhibition group. Alkaline phosphatase (ALP) staining was used to identify osteogenesis, alizarin red staining was used to detect calcium nodules precipitation, and TUNEL was used to detect apoptosis. mRNA and protein expression levels were detected by Real-time PCR and Western blotting, and the potential binding sites of miR-30d-5p were predicted by the bioinformatics analysis website Targetscan 7.1. Results After miR-30d-5p overexpression, osteogenic differentiation ability, and mineralization ability of the cells decreased (P<0.05), while apoptosis level increased (P< 0.05). The expression of glucoregulatory protein 78 (GRP78) and osteogenic specific transcription factor Runt related transcription factor 2(RUNX2) decreased significantly (P<0.05). However, miR-30d-5p inhibitor-treated the cells with increased osteogenic differentiation and mineralization ability (P < 0.05), and apoptosis level decreased (P < 0.05). GRP78 and RUNX2 protein levels increased (P<0.05). The miR-30d-5p binding site was located at 142-148 bp of the 3'UTR of the GRP78 gene. Conclusion MiR-30d-5p inhibits osteogenic differentiation and promotes apoptosis of bone marrow stromal cells by down-regulating the expression of GRP78 protein.
ABSTRACT
Objective To investigate the diagnosis and prognosis value of plasma microRNA-30d (miR-30d)in acute coronary syndrome(ACS)patients.Methods It retrospectively recruited 170 cases of ACS patients from TEDA International Cardiovascular Hospital between September 2011 to February 2012, including 70 STEMI(male 54,female 16), 52 NSTEMI(male 34,female 18),48 UAP(male 29,female 19).At the same time,41 healthy controls(male 24,female 17)were enrolled into the study.Plasma miR-30d levels were determined by real-time quantitative PCR.In order to evaluate the dynamic change of miR-30d and other cardiac biomarkers,20 plasma samples of AMI patients were collected at 0-3 h,4-6 h,7-9 h, 10-12 h after pectoralgia.ROC curves and Kaplan-Meier survival curve were used to investigate clinical value of miR-30d in ACS.Results At 0-3 h after pectoralgia, miR-30d were significant higher in STEMI 7.208(0.170-11 070.735)and NSTEMI 7.989(0.836-151.391)than the controls 1.561(0.044-17.520)（Z1＝-5.792,Z2 ＝-6.113,P＜0.001）, but there were no statistic differences between UAP 1.073 （0.051-11.095）patients and the controls（Z＝-0.325,P＝0.745）.In 20 AMI patients,miR-30d levels peaked at 4-6 h and then dropped following 7-9 h, both earlier than cTnI, and the variation tendency was positive correlated with cTnI（r＝0.402,P＜0.01）.At 0-3 h after pectoralgia, the AUC, sensitivity and specificity of miR-30d for differentiating AMI and UAP were 0.882（95％ CI:0.830-0.935）,0.795（95％CI:0.711-0.861）and 0.854（95％ CI:0.716-0.935）respectively.When combined miR-30d and cTnI, the diagnostic AUC and specificity were 0.937（95％ CI: 0.902-0.972）and 0.937（95％ CI:0.818-0.984）,both enhanced when compared with miR-30d or cTnI alone.Kaplan-Meier survival curves revealed that there were no significant correlations between the miR-30d levels and MACE in both 30 days and 12 months（χ$lt@span sup=1$gt@2$lt@/span$gt@＝0.506,P＝0.477 and χ$lt@span sup=1$gt@2$lt@/span$gt@＝0.002, P＝0.963 respectively）.Conclusion Plasma miR-30d may be used as a potential biomarker for early diagnosis, but not prognosis in ACS patients.