Inhibition Effect of Physalis angulata Leaf Extract on Viability, Collagen Type I, and Tissue Inhibitor of Metalloproteinase 1 (TIMP-1) but Not Plasminogen Activator Inhibitor-1 (PAI-1) of Keloid Fibroblast Culture

Abstract

Introduction: Keloids are excessive fibroproliferative diseases that are caused by abnormal wound healing. The anti-proliferative activity of Physalis angulata compounds has potential as a keloid therapeutic agent. This study aimed to observe the effects of P. angulata on fibroblast viability and collagen type I, tissue inhibitor of metalloproteinase 1 (TIMP-1), and plasminogen activator inhibitor 1 (PAI-1) levels in human keloid fibroblasts.

Methods: We conducted an experimental study of P. angulata ethanol extract on three primary human keloid fibroblast 3 passage cultures with four replications. Fibroblast viability was measured using the MTT assay after incubation with 3, 5, and 10 µg/mL P. angulata. Concentrations of P. angulata used to observe effects on TIMP-1, PAI-1, and collagen type I levels were 10%, 20%, 30%, and 40% of inhibitory concentration 50 (IC₅₀). The levels of collagen type I, TIMP-1, and PAI-1 were measured by ELISA. Mean comparisons between multiple treatment groups were analyzed using one-way ANOVA followed by post-hoc analysis.

Results: The 10 µg/mL P. angulata group had significantly lower fibroblast viability than the control group (p<0.05) with an IC₅₀ 6.3 µg/mL. The collagen type I level of 10% IC₅₀ (0.63 µg/mL) P. angulata group was lower than control (12.910 vs 47.866 ng/mL) (p=0.042). Level of TIMP-1 in 40% IC₅₀ (2.51 µg/mL) P. angulata group was lower than control (5.350 vs 9.972 ng/mL) (p=0.043). There was no significant difference in the PAI-1 levels.

Conclusion: This study showed the inhibitory effect of 10 µg/mL P. angulata extract on keloid fibroblast viability, with an IC₅₀ of 6.3 µg/mL. This study also showed collagen type-1 and TIMP-1 inhibition, but not PAI-1 inhibition, after P. angulate treatment.

Keywords: PAI-1; Physalis angulata; TIMP-1; collagen type I; fibroblast viability; keloid.

Figure 1

Keloid fibroblast culture. (A) One day after culture procedure. (B) First passage. (C) Second passage. (D) Third passage. Bar: 200 μm.

Figure 2

Formazan purple formation in MTT assay. (A) Control group. (B) Group of 3 μg/mL P. angulata extract. (C) Group of 5 μg/mL P. angulata extract. (D) Group of 10 μg/mL P. angulata extract showed the least formazan purple formation. Bar: 200 μm.

Figure 3

Effect of Physalis angulata extract on fibroblast viability. Keloid fibroblasts were incubated with 3, 5, and 10 µg/mL P. angulata extract. Control group incubated with culture media. Incubation time was 24 h. *: Significant fibroblast viability inhibition was showed at 10 µg/mL P. angulata group (p=0.023).

Figure 4

Effect of Physalis angulata extract on collagen type I. Keloid fibroblasts were incubated with 0.63 (10% IC₅₀), 1.25 (20% IC₅₀), 1.88 (30% IC₅₀), and 2.51 (40% IC₅₀) µg/mL P. angulata extract. Control group incubated with culture media. Incubation time was 24 h. *: Significant collagen type I inhibition was showed at 0.63 µg/mL (10% IC₅₀) P. angulata group (p=0.004).

Figure 5

Effect of Physalis angulata extract on TIMP-1. Keloid fibroblasts were incubated with 0.63 (10% IC₅₀), 1.25 (20% IC₅₀), 1.88 (30% IC₅₀), and 2.51 (40% IC₅₀) µg/mL P. angulata extract. Control group incubated with culture media. Incubation time was 24 h. *: Significant TIMP-1 inhibition was showed at 2.51 µg/mL (40% IC₅₀) P. angulata group. p=0.043.

Figure 6

Effect of Physalis angulata extract on PAI-1. Keloid fibroblasts were incubated with 0.63 (10% IC₅₀), 1.25 (20% IC₅₀), 1.88 (30% IC₅₀), and 2.51 (40% IC₅₀) µg/mL P. angulata extract. Control group incubated with culture media. Incubation time was 24 h. There were no significant different at PAI-1 level between all groups by Kruskal–Wallis test.