ABSTRACT
Objective To investigate the effect of pathologically elevated cyclic strain induced by hypertension on proliferation of vascular smooth muscle cells (VSMCs) and the role of long non-coding RNA (IncRNA)-XR007793 during this process. Methods Flexcell-4000 tension system was used to apply physiologically (5% magnitude) and pathologically (15% magnitude) cyclic strain with frequency of 1.25 Hz on VSMCs for 24 h respectively. qRT-PCR was used to detect the expression of XR007793 and 4 co-expressed genes: signal transducer and activator of transcription 2 (STAT2), cell division cycle associated 8 (CDCA8), proto-oncogene LMO2 and interferon regulatory factor (IRF7). Western blot was used to detect the proliferating cell nuclear antigen (PCNA) level in VSMCs. RNA inference was used to inhibit XR007793 expression. The cell cycle of VSMCs was measured by flow cytometry in static condition and the cell proliferation was detected by Brdu-Elisa in cyclic strain loading condition. Results Compared with 5% cyclic strain, 15% cyclic strain remarkably decreased the XR007793 level and increased the proliferation of VSMCs，along with the increasing expression of STAT2 and CDCA8. XR007793 specific siRNA transfection under static condition decreased the expression of XR007793 and increased the VSMC proliferation. Under 15% cyclic strain, XR007793 specific siRNA transfection also increased the VSMC proliferation and the expression of CDCA8 compared with the non-specific siRNA control. Conclusions Pathologically elevated cyclic strain decreases the XR007793 expression level and increases the CDCA8 expression level to modulate VSMC proliferation. These results provide new experimental evidence for the study of mechanobiological mechanism during hypertension and potential targets for hypertension therapy.
ABSTRACT
Objective To investigate the role of microRNA-34a (miR-34a) in the proliferation of vascular smooth muscle cells (VSMCs) induced by low shear stress (LowSS). Methods Using co-culture parallel plate flow chamber system, endothelial cells (ECs) and VSMCs were co-cultured and applied with normal shear stress (1.5 Pa) and LowSS (0.5 Pa) for 12 h. The expression of proliferating cell nuclear antigen (PCNA) in the co-cultured VSMCs was detected by Western blotting to determine the proliferation capacity of VSMCs. Real-time PCR was used to examine the miR levels of miR-34a in the co-cultured VSMCs. The target proteins of miR-34a were predicted by TargetScan, miRWalk and some other websites. Western blotting was used to detect expression of Forkhead box j2 (Foxj2) in the co-cultured VSMCs. Mimics and inhibitor were used to up-regulate or inhibit the expression of miR-34a, and then the expression of Foxj2 and PCNA was detected by Western blotting to verify the regulation relationship between miR 34a and Foxj2. Results Compared with NSS, LowSS promoted the PCNA expression and significantly up-regulated the miR-34a expression in the co-cultured VSMCs. Foxj2 was predicted to be the downstream target protein of miR-34a by TargetScan, miRWalk and some other websites. Foxj2 expression decreased significantly in the co-cultured VSMCs under LowSS application. Under static condition, the expression of Foxj2 obviously decreased and the expression of PCNA obviously increased by up-regulating miR-34a expression in VSMCs. While inhibiting the expression of miR-34a in VSMCs would result in a significant increase in the expression of Foxj2 and a significant decrease in the expression of PCNA. Conclusions LowSS can promote the proliferation of VSMCs by regulating miR-34a and target protein Foxj2 in the co-cultured VSMCs. This research finding will provide new mechanobiological experimental reference for further illustrating the pathogenesis of atherosclerosis and finding the therapeutic targets for drugs.
ABSTRACT
<p><b>OBJECTIVE</b>To assess bone health in epileptic children who have been treated with topiramate (TPM) or carbamazepine (CBZ).</p><p><b>METHODS</b>Sixty-three epileptic children who received TPM or CBZ treatment and 36 eileptic children who did not receive any drug treatment (control group) were enrolled. Bone mineral density (BMD) at lumbar vertebrae (L1-L4) and radius-ulna was evaluated by the dual-energy X-ray absorptiometry method. Biochemical indices of bone metabolism, including serum calcium, phosphorus and alkaline phosphatase contents were measured.</p><p><b>RESULTS</b>The serum calcium content was higher in the TPM group (2.41+/-0.17 mmol/L), but it was lower in the CBZ group (2.15+/-0.26 mmol/L) than that (2.26+/-0.11 mmol/L) in the control group (p<0.05). The serum phosphorus content in both the TPM (1.55+/-0.17 mmol/L) and the CBZ groups (1.52+/-0.26 mmol/L) was significantly lower than that in the control group (1.70+/-0.30 mmol/L) (p<0.05). There were no significant differences in the serum content of alkaline phosphatase between three groups. BMD was significantly reduced in both the TPM and the CBZ groups when compared to the control group (p<0.05).</p><p><b>CONCLUSIONS</b>TPM and CBZ may result in alterations in serum contents of calcium, phosphorus and alkaline phosphatase as well as BMD reduction.</p>