Sains Malaysiana 51(8)(2022): 2595-2608

http://doi.org/10.17576/jsm-2022-5108-19

Kesan Rawatan Krim Ekstrak Air Kacip Fatimah (Labisia pumila) terhadap Analisis Gen yang Terlibat dengan Proses Penyembuhan Luka melalui Kajian in vitro Model Kulit Manusia 3D dan in vivo Model Tikus

(Effects of Kacip Fatimah (Labisia pumila) Water Extract Cream on Gene Analysis Involved in Wound Healing Process Through the Study of in vitro 3D Human Skin Model and in vivo Rat Model)

SITI MUNEERAH MOHD ABD RAHMAN¹, AFIQAH SHAFIFY AMRAN¹, NUR SHAZWANI MOHD PILUS¹, ISA NAINA MOHAMED² & NURUL YUZIANA MOHD YUSOF^1,*

¹Jabatan Sains Bumi dan Alam Sekitar, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia

²Jabatan Farmakologi, Fakulti Perubatan, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, Bandar Tun Razak, 56000 Kuala Lumpur, Wilayah Persekutuan, Malaysia

Received: 10 December 2021/Accepted: 17 February 2022

Abstrak

Kajian ini melaporkan kesan rawatan bagi dua sediaan krim yang masing-masing mengandungi 1% ekstrak air bagi dua varieti L. pumilaiaitu var. alata dan var. pumila terhadap penyembuhan luka melalui uji kaji menggunakan model tikus dan model kulit manusia 3D EpidermFull Thickness (Mat Tek, USA) melalui penentuan penutupan luka dan analisis pengekspresan gen terpilih bagi mekanisme penyembuhan luka tisu kulit (gen TGF-1, IL-6, EGF, CBL dan COL3A1 untuk tikus; gen IL-10, TGFb-1 dan COL3A1 untuk manusia) menggunakan pendekatan tindak balas berantai polimerase kuantitatif (qPCR). Selepas rawatan, kawasan luka pada kulit bahagian atas badan tikus diukur pada hari ke-2, ke-5 dan ke-8, serta disampel untuk analisis pengekspresan gen. Bagi uji kaji in vitro tisu binaan kulit manusia, perubahan terhadap setiap lapisan luka yang dibuat pada binaan tisu kulit tersebut dicerap melalui pemerhatian histologi dan dianalisis bagi profil pengekspresan gen berdasarkan sela hari ke-2, ke-5 dan ke-6. Rawatan krim ekstrak air L. pumilakedua-dua varieti didapati hanya berkesan membantu penutupan luka bagi tikus betina iaitu 79.58% bagi var. alata pada hari ke-5 rawatan dan 75.97 dan 95.04% bagi var. pumila masing-masing pada hari ke-5 dan ke-8 rawatan tetapi tidak berkesan kepada tikus jantan. Namun begitu, dari segi tempoh penutupan luka lengkap, didapati kesemua rawatan menunjukkan luka ditutup lebih cepat berbanding kawalan (17 hari) iaitu di antara 12 sehingga 13 hari. Analisis gen bagi sampel luka tikus menunjukkan peningkatan aras gen hanya melibatkan gen IL-6 untuk tikus jantan yang dirawat dengan L. pumilavar. alata iaitu sebanyak 1722784 salinan mRNA pada hari ke-2 rawatan, gen CBL untuk tikus betina yang dirawat dengan L. pumilavar. alata berjumlah 46137 salinan mRNA pada hari ke-5, dan gen COL3A1 bagi rawatan L. pumilavar. alata untuk kedua-dua kumpulan jantina, iaitu masing-masing 2.44×10⁸ salinan mRNA (betina) dan 3.91×10⁸ salinan mRNA (jantan). Berdasarkan pemerhatian histologi sampel luka model kulit manusia 3D, rawatan kedua-dua varieti L. pumila menunjukkan kesan pengunjuran sel daripada kawasan luka pertumbuhan tetapi analisis gen tidak menunjukkan perubahan yang tidak signifikan terhadap gen kajian. Secara keseluruhannya, hasil kajian ini dapat menunjukkan potensi L. pumiladalam penyembuhan luka tetapi perlu dimantapkan lagi pada masa hadapan melalui strategi uji kaji in vivo dan in vitro yang lebih optimum.

Kata kunci: Asai penyembuhan luka; ekstrak air Labisia pumila; model tisu bersamaan kulit manusia

Abstract

This study reported the treatment effects of two cream preparations each containing 1% water extract for two variations of L. pumila namely var. alata dan var. pumila on wound healing based on experiments using rat model and three dimensional (3D) human skin model, Epiderm Full Thickness (Mat Tek Corporation, USA) through wound closure assessment and analysis of selected gene expression for skin tissue wound healing mechanism (TGF-1, IL-6, EGF, CBL genes and COL3A1 for rats; the genes IL-10, TGFb-1 and COL3A1 for humans) using a quantitative polymerase chain reaction (qPCR) approach. After treatment, the lesion area on the skin of the upper body of the rats was measured on days 2, 5 and 8, as well as sampled for gene expression analysis. For in vitro experiments of human skin tissue, changes to each layer of lesion made on the skin tissue were observed through histological observations and analyzed for gene expression profiles based on 2nd, 4th, and 6th day intervals. Water extract treatments of L. pumila of both variations was found to be only effective in helping wound closure for female rats at 79.58% for var. alata on the 5th day of treatment and 75.97 and 95.04% for var. pumila on the 5th and 8th day of treatment, respectively, but was ineffective in male rats. However, in terms of the period of complete wound closure, it was found that all treatments showed that the wound closed faster than the control (17 days), which is between 12 and 13 days. Gene analysis for rat wound samples showed increased gene levels involving only the IL-6 gene for male rats treated with L. pumila var. alata with a total of 1722784 messenger ribonucleic acid (mRNA) copy number on day 2 of treatment, the CBL gene for female mice treated with L. pumila var. alata totaled 46137 mRNA copy number on day 5, and the COL3A1 gene for the treatments of L. pumila var. alata for both sex groups, with total of 2.44×10⁸ mRNA copy number (females) and 3.91×10⁸ mRNA copy number (males), respectively. Based on histological observation of human skin tissue-constructed wound samples, treatments of both variants of L. pumila showed a cell projection effect from the growth wound area but gene analysis showed insignificant changes to the study genes. Overall, the results of this study can indicate the potential of L. pumila in wound healing but needs to be further strengthened in the future through more optimal in vivo and in vitro experimental strategies.

Keywords: Human skin equivalent tissue model; Labisia pumilawater extract; wound healing assay

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^*Corresponding author; email: yuziana@ukm.edu.my