A novel dimeric thymosin beta 4 with enhanced activities accelerates the rate of wound healing.
A novel dimeric thymosin beta 4 with enhanced activities accelerates the rate of wound healing.
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DOI：
10.2147/dddt.s50183
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Li M
中科院分区：
文献类型：
--
作者：
Xu TJ;Wang Q;Ma XW;Zhang Z;Zhang W;Xue XC;Zhang C;Hao Q;Li WN;Zhang YQ;Li M
胸腺素β 4（Tβ4）是一种具有43个氨基酸的肽，对于皮肤，眼睛，心脏和神经系统损伤后的修复和重塑组织至关重要。为了充分实现其作为损伤引起的疾病的治疗的效用，作者构建了一种具有成本效益的新型Tβ4二聚体，并证明它能够比天然Tβ4更好地加速组织修复。构建了含有两个完整Tβ4基因的原核表达载体，并在大肠杆菌中表达。采用10 L中试规模发酵生产工程菌，通过一步疏水层析法纯化Tβ4二聚体。通过四甲基联苯胺（甲基噻唑四唑）、trans-well、划痕和管形成试验评估Tβ4二聚体促进内皮细胞增殖、迁移和发芽的活性。在大鼠上评估加速皮肤愈合的能力。发酵后，Tβ4二聚体约占菌体总蛋白的30%。经疏水层析纯化后，Tβ4二聚体的纯度可达98%。在10 L发酵罐中，Tβ4二聚体的平均产量为562.4mg/L。在每项试验中，与天然Tβ4相比，二聚体Tβ4表现出增强的活性。值得注意的是，二聚体Tβ4促进细胞迁移的能力几乎是Tβ4的两倍。二聚体Tβ4处理大鼠的皮肤愈合速率比天然Tβ4处理大鼠快约1天。与天然Tβ4相比，二聚体Tβ4具有更强的伤口愈合活性，且纯化工艺简单，成本低。该数据对于与慢性伤口疾病相关的高疼痛和发病率可能具有显著益处。还描述了一种更好的策略，即开发Tβ4作为治疗心脏病发作、神经营养性角膜炎和多发性硬化等损伤引起的其他疾病的治疗方法。
Thymosin beta 4 (Tβ4) is a peptide with 43 amino acids that is critical for repair and remodeling tissues on the skin, eye, heart, and neural system following injury. To fully realize its utility as a treatment for disease caused by injury, the authors constructed a cost-effective novel Tβ4 dimer and demonstrated that it was better able to accelerate tissue repair than native Tβ4. A prokaryotic vector harboring two complete Tβ4 genes with a short linker was constructed and expressed in Escherichia coli. A pilot-scale fermentation (10 L) was performed to produce engineered bacteria and the Tβ4 dimer was purified by one-step hydrophobic interaction chromatography. The activities of the Tβ4 dimer to promote endothelial cell proliferation, migration, and sprouting were assessed by tetramethylbenzidine (methylthiazol tetrazolium), trans-well, scratch, and tube formation assays. The ability to accelerate dermal healing was assessed on rats. After fermentation, the Tβ4 dimer accounted for about 30% of all the bacteria proteins. The purity of the Tβ4 dimer reached 98% after hydrophobic interaction chromatography purification. An average of 562.4 mg/L Tβ4 dimer was acquired using a 10 L fermenter. In each assay, the dimeric Tβ4 exhibited enhanced activities compared with native Tβ4. Notably, the ability of the dimeric Tβ4 to promote cell migration was almost two times higher than that of Tβ4. The rate of dermal healing in the dimeric Tβ4-treated rats was approximately 1 day faster than with native Tβ4-treated rats. The dimeric Tβ4 exhibited enhanced activity on wound healing than native Tβ4, and the purification process was simple and cost-effective. This data could be of significant benefit for the high pain and morbidity associated with chronic wounds disease. A better strategy to develop Tβ4 as a treatment for other diseases caused by injuries such as heart attack, neurotrophic keratitis, and multiple sclerosis was also described.