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Sains Malaysiana 51(8)(2022):
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 YUSOF1,*
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
Diserahkan: 10 Disember 2021/Diterima:
17 Februari 2022
Abstrak
Kajian ini melaporkan kesan rawatan bagi dua sediaan krim yang masing-masing mengandungi 1% ekstrak air bagi dua varieti L. pumila iaitu 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. pumila kedua-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. pumila var. 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 pumila water extract; wound healing
assay
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*Pengarang untuk surat-menyurat;
email: yuziana@ukm.edu.my
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