Lentiviral expression vectors for calcitonin gene-related peptide (CGRP) were used to transfect rat bone marrow mesenchymal stem cells (MSCs). After assessing the biological characteristics of proliferation and aging in MSCs transfected with CGRP, we observed the effects of the CGRP-modified rat MSCs on the migration and proliferation of rat vascular smooth muscle cells (VSMCs) in vitro. Rat MSCs were isolated, cultured in vitro, and identified by flow cytometry. A CGRP recombinant lentivirus was transfected into MSCs. The transfection efficiency was determined by fluorescence microscopy and flow cytometry, and CGRP in MSCs was detected by real-time quantitative PCR, ELISA, and immunofluorescence. The proliferation and senescence of CGRP-modified MSCs were evaluated by MTT assay and beta-galactosidase staining. VSMCs were isolated, cultured in vitro, and identified by immunofluorescence. CGRP-modified MSCs and VSMCs were cocultured in a Transwell system. The proliferation and migration of VSMCs were evaluated by scratch testing and the MTT method. Rat bone marrow MSCs showed a spindle-shaped morphology, adherent growth in vitro, positive CD29 and CD90 expression, and negative CD45 expression. CGRP was stably expressed in MSCs after 48 h of recombinant lentivirus transfection. CGRP mRNA and protein secretion in CGRP recombinant lentivirus-transfected MSCs were higher than that in control MSCs. Immunofluorescence showed that CGRP protein could be expressed in CGRP-modified MSCs. The proliferation ability and senescence rates did not differ between lentivirus-transfected MSCs and untransfected MSCs. Rat VSMCs expressed α-SMA protein and exhibited a spindle-shaped morphology and adherent growth in vitro. In Transwell coculture experiments, scratch testing of VSMCs showed that CGRP-modified MSCs could reduce VSMC proliferation and migration. The CGRP gene can be stably expressed in MSCs after CGRP recombinant lentivirus transfection. CGRP recombinant lentivirus transfection has little effect on the proliferation or senescence of MSCs, and CGRP-modified MSCs can inhibit the proliferation and migration of VSMCs. These results lay a foundation for research on the use of CGRP gene-engineered MSCs in restenosis therapy.

降钙素基因相关肽（CGRP）的慢病毒表达载体用于转染大鼠骨髓间充质干细胞（MSC）。评估增殖的生物学特性，并与CGRP转染的MSC老化后，我们观察到对迁移和大鼠增殖的CGRP改性大鼠MSC的影响血管平滑肌细胞（VSMCs）体外。分离大鼠MSC ，体外培养，并通过流式细胞仪鉴定。将CGRP重组慢病毒转染到MSC中。通过荧光显微镜和流式细胞术确定转染效率，并通过实时定量PCR，ELISA和免疫荧光检测MSC中的CGRP。通过MTT测定和β-半乳糖苷酶染色评估了CGRP修饰的MSC的增殖和衰老。分离VSMC，在体外培养，并通过免疫荧光鉴定。CGRP修饰的MSC和VSMC在Transwell系统中共培养。通过划痕试验和MTT法评估了VSMC的增殖和迁移。大鼠骨髓间充质干细胞呈纺锤形形态，体外黏附生长，CD29和CD90阳性表达和CD45阴性表达。重组慢病毒转染48小时后，CGRP在MSC中稳定表达。CGRP重组慢病毒转染的MSCs中的CGRP mRNA和蛋白分泌高于对照MSCs。免疫荧光显示，CGRP蛋白可以在CGRP修饰的MSC中表达。慢病毒转染的MSCs和未转染的MSCs的增殖能力和衰老率没有差异。大鼠VSMC表达α-SMA蛋白并在体外表现出纺锤形形态和粘附生长。在Transwell共培养实验中，对VSMC的划痕测试表明，CGRP修饰的MSC可以减少VSMC的增殖和迁移。CGRP重组慢病毒转染后，CGRP基因可以在MSC中稳定表达。CGRP重组慢病毒转染对MSC的增殖或衰老影响很小，而CGRP修饰的MSC可以抑制VSMC的增殖和迁移。这些结果为研究CGRP基因工程的MSC在再狭窄治疗中的应用奠定了基础。