PDF(Pubmed)
Abstract：
BACKGROUND: Skin fibrosis affects the normal function of the skin. TGF-β1 is a key cytokine that affects organ fibrosis. The latency-associated peptide (LAP) is essential for TGF-β1 activation. We previously constructed and prepared truncated LAP (tLAP), and confirmed that tLAP inhibited liver fibrosis by affecting TGF-β1. SPACE peptide has both transdermal and transmembrane functions. SPACE promotes the delivery of macromolecules through the stratum corneum into the dermis. This study aimed to alleviate skin fibrosis through the delivery of tLAP by SPACE.
METHODS: The SPACE-tLAP (SE-tLAP) recombinant plasmid was constructed. SE-tLAP was purified by nickel affinity chromatography. The effects of SE-tLAP on the proliferation, migration, and expression of fibrosis-related and inflammatory factors were evaluated in TGF-β1-induced NIH-3T3 cells. F127-SE-tLAP hydrogel was constructed by using F127 as a carrier to load SE-tLAP polypeptide. The degradation, drug release, and biocompatibility of F127-SE-tLAP were evaluated. Bleomycin was used to induce skin fibrosis in mice. HE, Masson, and immunohistochemistry were used to observe the skin histological characteristics.
RESULTS: SE-tLAP inhibited the proliferation, migration, and expression of fibrosis-related and inflammatory factors in NIH-3T3 cells. F127-SE-tLAP significantly reduced ECM production, collagen deposition, and fibrotic pathological changes, thereby alleviating skin fibrosis.
CONCLUSIONS: F127-SE-tLAP could increase the transdermal delivery of LAP, reduce the production and deposition of ECM, inhibit the formation of dermal collagen fibers, and alleviate the progression of skin fibrosis. It may provide a new idea for the therapy of skin fibrosis.
METHODS: The SPACE-tLAP (SE-tLAP) recombinant plasmid was constructed. SE-tLAP was purified by nickel affinity chromatography. The effects of SE-tLAP on the proliferation, migration, and expression of fibrosis-related and inflammatory factors were evaluated in TGF-β1-induced NIH-3T3 cells. F127-SE-tLAP hydrogel was constructed by using F127 as a carrier to load SE-tLAP polypeptide. The degradation, drug release, and biocompatibility of F127-SE-tLAP were evaluated. Bleomycin was used to induce skin fibrosis in mice. HE, Masson, and immunohistochemistry were used to observe the skin histological characteristics.
RESULTS: SE-tLAP inhibited the proliferation, migration, and expression of fibrosis-related and inflammatory factors in NIH-3T3 cells. F127-SE-tLAP significantly reduced ECM production, collagen deposition, and fibrotic pathological changes, thereby alleviating skin fibrosis.
CONCLUSIONS: F127-SE-tLAP could increase the transdermal delivery of LAP, reduce the production and deposition of ECM, inhibit the formation of dermal collagen fibers, and alleviate the progression of skin fibrosis. It may provide a new idea for the therapy of skin fibrosis.
摘要：
背景：皮肤纤维化影响皮肤的正常功能。TGF-β1是影响器官纤维化的关键细胞因子。潜伏期相关肽（LAP）对于TGF-β1激活至关重要。我们先前构建并准备了截断的LAP(tLAP)，并证实tLAP通过影响TGF-β1抑制肝纤维化。SPACE肽具有经皮和跨膜功能。SPACE促进大分子通过角质层进入真皮。本研究旨在通过SPACE递送tLAP来缓解皮肤纤维化。
方法：构建SPACE-tLAP(SE-tLAP)重组质粒。SE-tLAP通过镍亲和层析纯化。SE-tLAP对细胞增殖的影响,迁移,并观察TGF-β1诱导的NIH-3T3细胞中纤维化相关因子和炎症因子的表达。以F127为载体负载SE-tLAP多肽,构建F127-SE-tLAP水凝胶。退化，药物释放，并对F127-SE-tLAP的生物相容性进行了评价。博来霉素用于诱导小鼠皮肤纤维化。他,Masson,用免疫组化方法观察皮肤组织学特征。
结果：SE-tLAP抑制细胞增殖，迁移,NIH-3T3细胞中纤维化相关因子和炎症因子的表达。F127-SE-tLAP显著降低ECM产量，胶原蛋白沉积，和纤维化病理变化，从而减轻皮肤纤维化。
结论：F127-SE-tLAP可以增加LAP的透皮给药，减少ECM的产生和沉积，抑制真皮胶原纤维的形成，并缓解皮肤纤维化的进展。为皮肤纤维化的治疗提供了新的思路。
方法：构建SPACE-tLAP(SE-tLAP)重组质粒。SE-tLAP通过镍亲和层析纯化。SE-tLAP对细胞增殖的影响,迁移,并观察TGF-β1诱导的NIH-3T3细胞中纤维化相关因子和炎症因子的表达。以F127为载体负载SE-tLAP多肽,构建F127-SE-tLAP水凝胶。退化，药物释放，并对F127-SE-tLAP的生物相容性进行了评价。博来霉素用于诱导小鼠皮肤纤维化。他,Masson,用免疫组化方法观察皮肤组织学特征。
结果：SE-tLAP抑制细胞增殖，迁移,NIH-3T3细胞中纤维化相关因子和炎症因子的表达。F127-SE-tLAP显著降低ECM产量，胶原蛋白沉积，和纤维化病理变化，从而减轻皮肤纤维化。
结论：F127-SE-tLAP可以增加LAP的透皮给药，减少ECM的产生和沉积，抑制真皮胶原纤维的形成，并缓解皮肤纤维化的进展。为皮肤纤维化的治疗提供了新的思路。
