[논문]The Effect of Trigonella foenum-graceum L. (Fenugreek) Towards Collagen Type I Alpha 1 (COL1A1) and Collagen Type III Alpha 1 (COL3A1) on Postmenopausal Woman's Fibroblast

Yusharyahya, Shannaz Nadia; Bramono, Kusmarinah; Sutanto, Natalia Rania; Kusuma, Indra

doi:10.20307/nps.2019.25.3.208

The Effect of Trigonella foenum-graceum L. (Fenugreek) Towards Collagen Type I Alpha 1 (COL1A1) and Collagen Type III Alpha 1 (COL3A1) on Postmenopausal Woman's Fibroblast 원문보기

Natural product sciences, v.25 no.3, 2019년, pp.208 - 214

Yusharyahya, Shannaz Nadia (Department of Dermatology and Venereology, Faculty of Medicine, Universitas Indonesia) , Bramono, Kusmarinah (Department of Dermatology and Venereology, Faculty of Medicine, Universitas Indonesia) , Sutanto, Natalia Rania (Department of Dermatology and Venereology, Faculty of Medicine, Universitas Indonesia) , Kusuma, Indra (Department of Physiology, Faculty of Medicine University of YARSI)

Abstract ▼ AI-Helper

Trigonella foenum-graceum L. (fenugreek) is a phytoestrogen, a nonsteroidal organic chemical compound from plants which has similar mechanism of action to sex hormone estradiol-$17{\beta}$. This study aims to assess the effectivity of fenugreek seeds extract on collagen type I alpha 1 (COL1A1) and collagen type III alpha 1 (COL3A1) which are both decreased in aging skin and become worsen after menopause. This in vitro experimental study used old human dermal fibroblast from leftover tissue of blepharoplasty on a postmenopausal woman (old HDF). As a control of the fenugreek's ability to trigger collagen production, we used fibroblast from preputium (young HDF). Subsequent to fibroblast isolation and culture, toxicity test was conducted on both old and young HDF by measuring cell viability on fenugreek extract with the concentration of 5 mg/mL to $1.2{\mu}g/mL$ which will be tested on both HDF to examine COL1A1 and COL3A1 using ELISA, compared to no treatment and 5 nM estradiol. Old HDF showed a 4 times slower proliferation compared to young HDF (p<0.05). Toxicity test revealed fenugreek concentration of $0.5-2{\mu}g/mL$ was non-toxic to both old and young HDF. The most significant fenugreek concentration to increase COL1A1 and COL3A1 secretion was $2{\mu}g/mL$ (p<0.05).

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문제 정의

There is no previous study evaluating the role of fenugreek seeds extract as anti-aging therapy for postmenopausal women. Based on the explanation above, this study aims to be the first to molecularly assess the role of fenugreek seeds extract effect towards COL1A1 and COL3A1.

제안 방법

Cells on 24 plates were harvested and viable cells were counted using trypan blue exclusion method by TC20 automated cell counter (Bio-Rad). A test was performed using COL1A1 (MyBioSource, USA, MBS763786) and COL3A1 (MyBioSource, USA, MBS763396) ELISA kit in duplo to confirm the results. The absorbance was measured according to protocol using multiplate reader (TECAN).
The patient was selected based on inclusion criteria: women who have been in menopause for at least 2 years and maximum 5 years, estradiol level <30 pg/mL, wrinkles on face and consented to donate their leftover skin tissue from surgery.

데이터처리

Statistical analysis − A statistic test was done using paired t-test with alpha value of 0.05 using Microsoft Excel 2016.

이론/모형

The conditioned medium sample obtained after 24 hours was centrifuged according to ELISA protocol instructions and stored at -80 oC. Cells on 24 plates were harvested and viable cells were counted using trypan blue exclusion method by TC20 automated cell counter (Bio-Rad). A test was performed using COL1A1 (MyBioSource, USA, MBS763786) and COL3A1 (MyBioSource, USA, MBS763396) ELISA kit in duplo to confirm the results.

성능/효과

In conclusion, fenugreek extract at the concentration of 2 μg/mL could significantly increase COL1A1 and COL3A1 secretion on postmenopausal woman’s fibroblast compared to no treatment and 5 nM estradiol controls.
Results of fenugreek extract toxicity test at 5 mg/mL to 9.8 μg/mL concentration showed that fenugreek extract was toxic to the old HDF.
The results from this study showed an increase of 1:1.5-2 in COL1A1 and COL3A1 ratio of old HDF after fenugreek exposure. Cheng³⁰ showed that COL1A1 and COL3A1 ratio is decreased as age increases.
This result showed that fenugreek's effect is not dose-dependent and that 2 μg/mL is the optimal concentration for COL1A1 and COL3A1 on both the old and the young HDF.

후속연구

In postmenopausal women, COL1A1 level is higher than COL3A1. Based on the results of this study, it is expected that using fenugreek as a postmenopausal skin aging therapy will produce a more significant increase of COL3A1 level compared to COL1A1.
No other study had tested fenugreek using old fibroblast as comparison. Therefore, further studies are still required to investigate about the exact mechanism of action of fenugreek whether it is through specific estrogen binding receptor or other pathways.
It could be concluded that fenugreek extract effect towards COL1A1 and COL3A1 is not dose-dependent. To find out the cause, further studies are needed to consider specificity of old fibroblast response to low concentration fenugreek extract, and that various active substances content of the extract could have different optimal concentration, therefore giving different cell responses. A study from Agustini¹² showed that fenugreek contains a lot of components, such as trigonelline, diosgenin, yamogenin, gitogenin, tiogenin, vitexin, isovitexin, quercetin, luteolin, orientin and isoorientin.

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연합인증

The Effect of Trigonella foenum-graceum L. (Fenugreek) Towards Collagen Type I Alpha 1 (COL1A1) and Collagen Type III Alpha 1 (COL3A1) on Postmenopausal Woman's Fibroblast 원문보기

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The Effect of Trigonella foenum-graceum L. (Fenugreek) Towards Collagen Type I Alpha 1 (COL1A1) and Collagen Type III Alpha 1 (COL3A1) on Postmenopausal Woman's Fibroblast 원문보기

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