Mechanism of the effect of Piper crocatum extract on wound healing of Wistar rats post-excision mammary tumor based on IL-10 level, TGF-β1 expression, VEGF expression, Collagen density, and clinical features

Abstract

Background: The incidence of wound complications after breast cancer surgery is > 30%. Delayed wound healing increases the risk of systemic recurrence up to three-fold after excisional surgery for primary breast cancer. Patients with breast malignant tumors exhibit immune system dysfunction while T cells play a major role in the inflammatory process of wound healing. The time required to achieve wound closure depends on the severity of the wound. The content of Piper crocatum ethanol extract contains alkaloids, flavonoids, tannins, steroids, and polyphenols. Flavonoid and polyphenol compounds are antioxidant, antidiabetic, anticancer, antiseptic, and anti-inflammatory.

Aim: To determine whether P. crocatum extract increases interleukin-10 (IL-10) levels, TGF-β1 and VEGF expression, collagen density, and the clinical features of wounds, and how the mechanism works on wound healing of Wistar rats after excision of mammary tumors.

Methods: This study used 35 female Wistar rats, 30 mammary tumor models by injecting 3-5% benzopyrene as much as 50 mg/Kg.BW in 5 injections into the mammary glands/mammae every 2 days as much as 10 mg/kg.BW. Thirty rats were divided into 5 groups; red betel extract gel 50%, 25%, and 12.5%; povidone iodine group as a positive control; and carboxymethyl cellulose natrium (CMC na) group as a negative control, 5 normal wounds as controls. Treatment was given 2 times a day for 14 days.

Results: Multiple linear regression results; P. crocatum extract decreased IL-10 levels by -0.64 (p = 0.025), decreased TGF-β1 expression -0.832 (p = 0.001), increased VEGF expression by 0.638 (p = 0.026), and decreased collagen density by -0.605 (p = 0.037) in mammary tumor post-excision rat wounds.

Conclusion: There is an effect of P. crocatum extract on the wound of Wistar rats after excision of mammary tumor by the mechanism of decreased IL-10 levels and TGF-β1 expression, increased VEGF expression, and decreased collagen density through increased VEGF expression is studied in this article.

Keywords: Collagen density; IL-10 level; Piper crocatum extract; TGF-β expression; VEGF expression.

Fig. 1.. Graph of the mean of IL-10 levels, TGF-β1 expression, VEGF expression, collagen density, and percentage of wound area in each group.

Fig. 2.. Histopathological features of the skin of Wistar rats post-excision of mammary tumors after treatment for 14 days in each group. TGF-β1 expression shown by macrophage cells (⇧) in Figure K (7.8), K- (7.4), K+ (5.7), P1 (1.7), P2 (4.7), and P3 (4) (HE staining, magnification of 400).

Fig. 3.. Histopathological features of Wistar rat skin post-excision of mammary tumor after treatment for 14 days in each group. VEGF expression shown in endothelial cells (⇧) in figure K (5.3), K- (6.1), K+ (8.5), P1 (7.5), P2 (13), and P3 (5.9) (HE staining, magnification of 400).

Fig. 4.. Histopathological features of Wistar rat skin post-excision of mammary tumors after treatment for 14 days in each group, as indicated by collagen (⇧) K (scoring 3) K- (scoring 0), K+ (scoring 1), P1 (scoring 2), P2 (scoring 1), and P3 (scoring 3) (HE staining, magnification of 400×).

Fig. 5.. Image J measurements on days 1^st, 7^th, and 14^th in each treatment group.

Fig. 6.. Appearance of wounds in the control group (A) and the Piper50% extract gel group (B) on the 14th day of treatment.

Fig. 7.. Path analysis of direct and indirect effects of Piper crocatum extract on IL-10 levels, VEGF expression, TGF-β1 expression, collagen density, and clinical features of post-excision mammary tumor wounds in female Wistar rats.

Fig. 8.. Mechanism of effect of Piper crocatum extract on IL-10 levels, TGF-β1 expression, VEGF expression, and collagen density in post-excision wound of mammary tumor of female Wistar rats.

Fig. 9.. Schematic representing the common growth factor signaling pathways that regulate key cellular processes required for wound healing and cancer (Sundaram et al., 2018).