Pentadecapeptide BPC 157 enhances the growth hormone receptor expression in tendon fibroblasts
Chung-Hsun Chang et al. Molecules. 2014.
Abstract
BPC 157, a pentadecapeptide derived from human gastric juice, has been demonstrated to promote the healing of different tissues, including skin, muscle, bone, ligament and tendon in many animal studies. However, the underlying mechanism has not been fully clarified. The present study aimed to explore the effect of BPC 157 on tendon fibroblasts isolated from Achilles tendon of male Sprague-Dawley rat. From the result of cDNA microarray analysis, growth hormone receptor was revealed as one of the most abundantly up-regulated genes in tendon fibroblasts by BPC 157. BPC 157 dose- and time-dependently increased the expression of growth hormone receptor in tendon fibroblasts at both the mRNA and protein levels as measured by RT/real-time PCR and Western blot, respectively. The addition of growth hormone to BPC 157-treated tendon fibroblasts dose- and time-dependently increased the cell proliferation as determined by MTT assay and PCNA expression by RT/real-time PCR. Janus kinase 2, the downstream signal pathway of growth hormone receptor, was activated time-dependently by stimulating the BPC 157-treated tendon fibroblasts with growth hormone. In conclusion, the BPC 157-induced increase of growth hormone receptor in tendon fibroblasts may potentiate the proliferation-promoting effect of growth hormone and contribute to the healing of tendon.
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Conflict of interest statement
The authors declare no conflict of interest.
Figures Figure 1
BPC 157 increased the expression of growth hormone receptor in tendon fibroblasts. Tendon fibroblasts at 50%–60% confluency were treated with BPC 157 at concentrations of 0, 0.1, 0.25, 0.5 μg/mL for 24 h (A and C) or 0.5 μg/mL for one to three days (B and D). The mRNA (A and B) and protein (C and D) expressions of growth hormone receptor were measured by RT/real-time PCR and Western blot analysis, respectively. Experiments were done in triplicate. Figure 2
Growth hormone increased the cell number of BPC 157-treated tendon fibroblasts. Tendon fibroblasts at 50%–60% confluency were pretreated with BPC 157 at concentrations of 0, 0.1, 0.25, 0.5 μg/mL for 24 h (A) or 0.5 μg/mL for one to three days (B). After BPC 157 pretreatment, 0.1 μg/mL growth hormone was added for another 24 h and then MTT assay was performed. Experiments were done in triplicate. The “★” would be applied if there is statistically significant. Figure 3
Growth hormone increased the PCNA gene expression in BPC 157-treated tendon fibroblasts. Tendon fibroblasts at 50%–60% confluency were pretreated with BPC 157 at concentrations of 0, 0.1, 0.25, 0.5 μg/mL for 24 h (A) or 0.5 μg/mL for one to three days (B). After BPC 157 pretreatment, 0.1 μg/mL growth hormone was added for another 24 h and then the PCNA gene expression was analyzed by RT/real-time PCR. Experiments were done in triplicate. The “★” would be applied if there is statistically significant. Figure 4
Growth hormone activated more JAK2 proteins in BPC 157-treated tendon fibroblasts. Tendon fibroblasts at 50%–60% confluency were pretreated with BPC 157 at concentration of 0.5 μg/mL for one to three days. After BPC 157 pretreatment, 0.1 μg/mL growth hormone was added for another 24 h and then total JAK2 (which was not shown) and activation of JAK2 which was determined by the level of phosphorylated JAK2 were detected by Western blot analysis (A). Experiments were done in triplicate. The levels of phosphorylated JAK2 were calculated by direct densitomeric analysis of the blot (B). The “★” would be applied if there is statistically significant.
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