PDF(Pubmed)
Abstract：
BACKGROUND: This study compared the differences of microvesicles (MVs) and microvesicles-delivering Smad7 (Smad7-MVs) on macrophage M1 polarization and fibroblast differentiation in a model of Peyronie\'s disease (PD).
METHODS: Overexpression of Smad7 in rat BMSCs was obtained by pCMV5-Smad7 transfection. MVs were collected from rat BMSCs using ultracentrifugation. In cells, 100 µg/mL of MVs or Smad7-MVs were used to treat the 100 ng/mL of lipopolysaccharide (LPS)-induced RAW264.7 cells or 10 ng/mL of recombinant transforming growth factor-β1 (TGF-β1)-induced fibroblasts. The pro-inflammatory cytokines and markers of M1 macrophages were measured in RAW264.7 cells, and the migration and markers of fibroblast differentiation were measured in fibroblasts. In rats, 50 µg of MVs or Smad7-MVs were used to treat the TGF-β1-induced animals. The pathology of tunica albuginea (TA), the markers of M1 macrophages and fibroblast differentiation in the TA were measured.
RESULTS: The MVs or Smad7-MVs treatment suppressed the LPS-induced macrophage M1 polarization and TGF-β1-induced fibroblast differentiation. Moreover, the Smad7-MVs treatment decreased the fibroblast differentiation compared with the MVs treatment. In the TGF-β1-induced TA of rats, MVs or Smad7-MVs treatment ameliorated the TA fibrosis by suppressing the macrophage M1 polarization and fibroblast differentiation. There was no significance on the M1-polarized macrophages between the MVs treatment and the Smad7-MVs treatment. Meanwhile, the Smad7-MVs treatment had an edge in terms of suppressing the fibroblast differentiation in the TGF-β1-induced PD model compared with the MVs treatment.
CONCLUSIONS: This study demonstrated that Smad7-MVs treatment had advantages over MVs treatment in suppressing of fibroblast differentiation in a model of PD.
METHODS: Overexpression of Smad7 in rat BMSCs was obtained by pCMV5-Smad7 transfection. MVs were collected from rat BMSCs using ultracentrifugation. In cells, 100 µg/mL of MVs or Smad7-MVs were used to treat the 100 ng/mL of lipopolysaccharide (LPS)-induced RAW264.7 cells or 10 ng/mL of recombinant transforming growth factor-β1 (TGF-β1)-induced fibroblasts. The pro-inflammatory cytokines and markers of M1 macrophages were measured in RAW264.7 cells, and the migration and markers of fibroblast differentiation were measured in fibroblasts. In rats, 50 µg of MVs or Smad7-MVs were used to treat the TGF-β1-induced animals. The pathology of tunica albuginea (TA), the markers of M1 macrophages and fibroblast differentiation in the TA were measured.
RESULTS: The MVs or Smad7-MVs treatment suppressed the LPS-induced macrophage M1 polarization and TGF-β1-induced fibroblast differentiation. Moreover, the Smad7-MVs treatment decreased the fibroblast differentiation compared with the MVs treatment. In the TGF-β1-induced TA of rats, MVs or Smad7-MVs treatment ameliorated the TA fibrosis by suppressing the macrophage M1 polarization and fibroblast differentiation. There was no significance on the M1-polarized macrophages between the MVs treatment and the Smad7-MVs treatment. Meanwhile, the Smad7-MVs treatment had an edge in terms of suppressing the fibroblast differentiation in the TGF-β1-induced PD model compared with the MVs treatment.
CONCLUSIONS: This study demonstrated that Smad7-MVs treatment had advantages over MVs treatment in suppressing of fibroblast differentiation in a model of PD.
摘要：
背景：这项研究比较了在佩罗尼病（PD）模型中，微泡（MV）和微泡递送Smad7（Smad7-MV）对巨噬细胞M1极化和成纤维细胞分化的差异。
方法：通过pCMV5-Smad7转染获得Smad7在大鼠BMSCs中的过表达。使用超速离心从大鼠BMSC收集MV。在细胞中，使用100μg/mL的MV或Smad7-MV治疗100ng/mL的脂多糖（LPS）诱导的RAW264.7细胞或10ng/mL的重组转化生长因子-β1（TGF-β1）诱导的成纤维细胞。在RAW264.7细胞中测量M1巨噬细胞的促炎细胞因子和标志物，并测量成纤维细胞的迁移和成纤维细胞分化标志物。在老鼠身上,使用50μg的MV或Smad7-MV治疗TGF-β1诱导的动物。白膜（TA）的病理学，测量了TA中M1巨噬细胞和成纤维细胞分化的标志物。
结果：MVs或Smad7-MVs处理抑制了LPS诱导的巨噬细胞M1极化和TGF-β1诱导的成纤维细胞分化。此外，与MV治疗相比,Smad7-MV治疗降低了成纤维细胞分化.在TGF-β1诱导的大鼠TA中,MV或Smad7-MV治疗通过抑制巨噬细胞M1极化和成纤维细胞分化来改善TA纤维化。MVs处理和Smad7-MVs处理之间的M1极化巨噬细胞没有显著性。同时,与MV治疗相比,Smad7-MV治疗在抑制TGF-β1诱导的PD模型中的成纤维细胞分化方面具有优势.
结论：本研究表明，在PD模型中，Smad7-MV治疗在抑制成纤维细胞分化方面优于MV治疗。
方法：通过pCMV5-Smad7转染获得Smad7在大鼠BMSCs中的过表达。使用超速离心从大鼠BMSC收集MV。在细胞中，使用100μg/mL的MV或Smad7-MV治疗100ng/mL的脂多糖（LPS）诱导的RAW264.7细胞或10ng/mL的重组转化生长因子-β1（TGF-β1）诱导的成纤维细胞。在RAW264.7细胞中测量M1巨噬细胞的促炎细胞因子和标志物，并测量成纤维细胞的迁移和成纤维细胞分化标志物。在老鼠身上,使用50μg的MV或Smad7-MV治疗TGF-β1诱导的动物。白膜（TA）的病理学，测量了TA中M1巨噬细胞和成纤维细胞分化的标志物。
结果：MVs或Smad7-MVs处理抑制了LPS诱导的巨噬细胞M1极化和TGF-β1诱导的成纤维细胞分化。此外，与MV治疗相比,Smad7-MV治疗降低了成纤维细胞分化.在TGF-β1诱导的大鼠TA中,MV或Smad7-MV治疗通过抑制巨噬细胞M1极化和成纤维细胞分化来改善TA纤维化。MVs处理和Smad7-MVs处理之间的M1极化巨噬细胞没有显著性。同时,与MV治疗相比,Smad7-MV治疗在抑制TGF-β1诱导的PD模型中的成纤维细胞分化方面具有优势.
结论：本研究表明，在PD模型中，Smad7-MV治疗在抑制成纤维细胞分化方面优于MV治疗。
