RESUMEN
MicroRNAs (miRNAs) have recently been shown to play a role in normal wound healing process. miRNAs may be linked to pathologic wound healing and closely related to the formation of hypertrophic scars. This study aimed to explore the effects of tetrandrine on the miRNA expression profile in human hypertrophic scar fibroblasts (HSFs) in vitro. HSFs were randomly divided into two groups: the tetrandrine treatment group and the control group. The experimental and control groups were collected and analyzed by miRNA array after a 48-h culture. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) was performed to confirm the array results. The targets of differentially expressed miRNA were functionally annotated using bioinformatic approaches. miRNA microarray analysis identified 193 differentially expressed miRNAs and the expression of 186 miRNAs in the experimental group decreased while that of 7 miRNAs increased compared to the control group. The most significantly downregulated miRNA was hsa-miR-1246, and hsa-miR-27b had the highest expression level. Significant differentially expressed miRNAs were predicted to be related to several important signaling pathways related to scar wound healing. The differential miRNA expression identified in this study provides the experimental basis for further understanding the anti-fibrosis effect of tetrandrine.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Bencilisoquinolinas/farmacología , Cicatriz Hipertrófica/genética , Fibroblastos/efectos de los fármacos , MicroARNs/genética , Cicatriz Hipertrófica/metabolismo , Cicatriz Hipertrófica/patología , Medicamentos Herbarios Chinos , Fibroblastos/metabolismo , Fibroblastos/patología , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Humanos , MicroARNs/agonistas , MicroARNs/antagonistas & inhibidores , MicroARNs/metabolismo , Anotación de Secuencia Molecular , Cultivo Primario de Células , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Despite years of effort, current therapies for diabetic wounds are still not fully efficacious. Emerging evidence has suggested that microRNAs (miRNAs) play key roles in multiple physiological and pathological processes in eukaryotes, and could potentially be powerful therapeutic tools. This study investigated the differential expression profiling of miRNAs in cutaneous wounds in streptozotocin-induced diabetic rats and normal rats, and its significance in diabetic wound healing. Using microarrays, 18 miRNAs were identified as being upregulated and 65 as being downregulated in the diabetic group. The miRNA profiling results were validated by quantitative reverse transcriptase polymerase chain reaction. Finally, functional annotation analysis using the DAVID and miR2Subpath databases revealed that the differentially expressed miRNAs were involved in MAPK signaling pathways, the Wnt signaling pathway, and other signaling pathways that may be closely linked to wound healing. This study provides an experimental foundation for further investigation of mechanisms that underlie poor diabetic wound healing, and of miRNA-based therapies that are associated with wound healing.
Asunto(s)
Complicaciones de la Diabetes , Diabetes Mellitus Experimental , MicroARNs/genética , Transcriptoma , Heridas y Lesiones/etiología , Animales , Biología Computacional , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Masculino , Anotación de Secuencia Molecular , Fenotipo , Ratas , Reproducibilidad de los Resultados , Transducción de Señal , Úlcera Cutánea/etiología , Úlcera Cutánea/genética , Úlcera Cutánea/metabolismo , Cicatrización de Heridas/genética , Heridas y Lesiones/genética , Heridas y Lesiones/metabolismoRESUMEN
We examined the protective effects of ultrafiltered XinMaiJia (XMJ) extract in a hydrogen peroxide (H2O2)-induced injury model in human umbilical vein endothelial cells (HUVECs) and determined the corresponding changes in the Na(+)-H(+) exchanger (NHE1) protein content and NHE1 gene expression. H2O2-induced HUVECs were treated with different concentrations of XMJ extract and the corresponding changes in morphology, activity, membrane permeability, biochemical indicators, cytokine concentration, NHE1 protein content, and NHE1 gene expression were determined. H2O2 significantly promoted HUVEC injury, whereas ultrafiltered XMJ extract significantly improved the morphological changes in injured HUVECs, increased their activity, and decreased NHE1 gene and protein expression, as well as limited the decrease in membrane permeability and expression of intercellular adhesion molecule-1, vascular cell adhesion molecule-1, interleukin (IL)-1, IL-6, and nuclear factor-kB. Ultrafiltered XMJ extract inhibited H2O2-induced HUVEC injury by inhibiting NHE1 activity.