Thymosin Beta-4 Recombinant Adeno-associated Virus Enhances Human Nucleus Pulposus Cell Proliferation and Reduces Cell Apoptosis and Senescence

Abstract

Background:Thymosin beta-4 (TB-4) is considered key roles in tissue development, maintenance and pathological processes. The study aimed to prove TB-4 positive biological function on nucleus pulposus (NP) cell apoptosis and slowing the process of cell aging while increasing the cell proliferation.Methods:TB-4 recombinant adeno-associated virus (AAV) was constructed and induced to human NP cells. Cell of same group were cultured without gene modification as controlled group. Proliferation capacity and cell apoptosis were observed during 6 passages of the cells. Morphology and expression of the TB-4 gene were documented as parameter of cell activity during cell passage.Results:NP cells with TB-4 transfection has normal TB-4 expression and exocytosis. NP cells with TB-4 transfection performed significantly higher cell activity than that at the control group in each generation. TB-4 recombinant AAV-transfected human NP cells also show slower cell aging, lower cell apoptosis and higher cell proliferation than control group.Conclusions:TB-4 can prevent NP cell apoptosis, slow NP cell aging and promote NP cell proliferation. AAV transfection technique was able to highly and stably express TB-4 in human NP cells, which may provide a new pathway for innovation in the treatment of intervertebral disc degenerative diseases.

Thymosin β4 suppresses CCl4‐induced murine hepatic fibrosis by down‐regulating transforming growth factor β receptor‐II

“…This requirement cannot be fulfilled using the usual commercially available vectors. In the process of constructing a recombinant AAV to mediate biologically active secretory expression of Tβ4, we used a novel secretory expression system in which the signal peptide and leader peptide of pro‐pre‐NT‐4 was fused to the N‐terminus of Tβ4 . This fusion pattern enabled the precise cleavage of the fusion protein to release Tβ4 with a Ser at its ultimate N‐terminus.…”

Progress on the Function and Application of Thymosin β4

“…Tb4 treatment alleviated tubular epithelial cell apoptosis by inhibiting the transforming growth factor (TGF)-b pathway in Sprague-Dawley (SD) rats with chronic renal tubular interstitial fibrosis (22). Moreover, it prevents nucleus pulposus cell apoptosis, reduces cellular aging, and promotes cell proliferation (23). Tb4 further decreased the apoptosis rate of EPCs induced by serum depletion and markedly downregulated the expression of the apoptosis-related proteins caspase-3 and caspase-9 in EPCs (24).…”

Anisodamine Maintains the Stability of Intervertebral Disc Tissue by Inhibiting the Senescence of Nucleus Pulposus Cells and Degradation of Extracellular Matrix via Interleukin-6/Janus Kinases/Signal Transducer and Activator of Transcription 3 Pathway

“…The staining was observed under a light microscope. Five high power fields were randomly selected, and the senescence process of HNPCs was determined by calculating the ratio of SA-β-gal positive cells to the total number of cells (Wang et al, 2015).…”