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Abstrak :

Menopause menyebabkan hipoestrogenisme dan mengakibatkan penuaan kulit. Fitoestrogen dari biji T. foenum-graecum (klabet) diharapkan dapat mengatasi penuaan kulit pascamenopause. Penelitian ini bertujuan untuk mengetahui konsentrasi optimal ekstrak klabet dalam menstimulasi sekresi COL1A1 dan COL3A1, cara kerjanya melalui jalur reseptor estrogen α (REα) atau REβ, dan peran klabet dalam mengurangi kerutan serta meningkatkan ketebalan kulit wajah.

Studi in vitro menggunakan human dermal fibroblast (HDF) tua yang diperoleh dari kultur sel fibroblas kulit perempuan pascamenopause dan HDF muda dari prepusium, dilakukan di Laboratorium Universitas YARSI. Sekresi COL1A1 dan COL3A1 diperiksa dengan ELISA lalu ditambah antagonis REa dan b. Penelitian in vivo merupakan uji klinis acak tersamar ganda berdurasi 12 minggu, di RSCM, Januari–November 2019. Subjek 50 perempuan pascamenopause yang dibagi dua: kelompok perlakuan mendapat krim klabet 5% dan kelompok plasebo mendapat krim dasar. Skor kerutan dahi, crow’s feet, dan nasolabial diukur dengan skoring atlas skin aging Bazin dan tebal dermis dengan high resolution ultrasound (HRU) 18 MHz.

Ekstrak klabet 2 µg/mL merupakan konsentrasi optimal dalam meningkatkan sekresi COL1A1 dan COL3A1 pada HDF tua dan muda dibandingkan tanpa perlakuan dan 5 nM estradiol. Uji inhibisi menunjukkan hambatan REα 50%; REβ dan RE α,β sampai 75%. Penurunan skor kerutan dahi, crow’s feet, dan nasolabial berbeda bermakna pada kedua kelompok minggu ke-4, ke-8, dan ke-12.  Peningkatan selisih skor kerutan dahi, crow’s feet, dan nasolabial tidak bermakna pada kedua kelompok antara baseline, minggu ke-4, ke-8, dan ke-12. Ketebalan dermis meningkat bermakna pada minggu ke-4 dibandingkan baseline. Pada minggu ke-8 dibandingkan minggu ke-4 dan pada minggu ke-12 dibandingkan minggu ke-8, ketebalan dermis menurun bermakna pada kedua kelompok. Peningkatan selisih ketebalan dermis pada minggu ke-4 dan ke-8 tidak bermakna.

Simpulan: Ekstrak klabet 2 µg/mL meningkatkan COL1A1 dan COL3A1 lebih banyak dibandingkan kelompok tanpa perlakuan dan estradiol 5 nM. Klabet bekerja terutama melalui REβ. Penurunan skor kerutan kulit dan peningkatan ketebalan kulit wajah tidak bermakna dibandingkan plasebo. Diduga, krim klabet konsentrasi 5% tidak tepat untuk mengurangi kerutan dan menambah ketebalan kulit. Perlu penelitian lebih lanjut untuk mengetahui konsentrasi optimal klabet sebagai terapi penuaan kulit wajah pascamenopause.

 

Kata kunci: COL1A1, COL3A1, kerutan kulit, ketebalan dermis, klabet, pascamenopause, penuaan kulit wajah, Trigonella foenum-graecum.

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Hypoestrogenism in menopause leads to skin aging, for which phytoestrogen originated from the seeds of T. foenum-graecum (fenugreek) is expected to be a solution. This study aims to show the effect of phytoestrogen from fenugreek extract in stimulating COL1A1 and COL3A1 through its mechanism of action on estrogen receptor (ER) ERa or ERb and its role in diminishing facial wrinkles and increasing dermal thickness.

The in vitro study was carried out in YARSI University Laboratory throughout March 2017–March 2018. This study looks into the effect of fenugreek extract on collagen level secretion in old and young human dermal fibroblast (HDF) compared to control. The in vivo study was a randomized, double-blind, 12-week trial conducted in RSCM from January–November 2019. Fifty postmenopausal women divided into two groups: the intervention group was given 5% fenugreek cream while the placebo group was given base cream.

The in vitro study showed that a concentration of 2 mg/mL was the optimal dose to stimulate COL1A1 and COL3A1 secretion in both old and young HDF compared to control (no treatment) and 5 nM estradiol. The inhibition test demonstrated suppression of ERa by 50%; ERb and ERa,b by up to 75%, indicating that the fenugreek activates both receptors, especially ERb. However, the success of the in vitro study did not translate into the in vivo study. Both the intervention group and the placebo group were able to achieve statistically significant in facial wrinkle scores from all focus areas without any significant disparity between both groups at all timepoints. Dermal thickness of facial skin showed similar results for both groups with significant improvements in the 8^thweek compared to baseline and significant decrease by the 12^thweek.

Conclusion: Fenugreek extract with a concentration of 2 mg/mL increased COL1A1 and COL3A1 secretion more potently compared to control and estradiol 5 nM. However, the decrease in facial skin wrinkles scores and the increase in dermal thickness were not significant compared to placebo. We suspect that a concentration of 5% was not adequate for the expected antiaging effects. Further studies are necessary to determine a more appropriate fenugreek concentration to permit clinical use as a postmenopausal antiaging therapy.

 

Keywords: COL1A1, COL3A1, dermal thickness, fenugreek, postmenopausal women, facial skin aging, skin wrinkles, Trigonella foenum-graecum.

Abstrak :
ABSTRAK
Prolaps uteri merupakan kondisi yang sering dialami oleh perempuan dan dapat menurunkan kualitas hidup. Penyebab prolaps uteri multifaktorial, pada umumnya berupa faktor klinis.Penelitian ini bertujuan untuk mengetahui adanya variasi gen berupa mutasi gen HOXA11 dan COL3A1 pada penderita prolaps uteri, mengetahui adanya perbedaan ekspresi protein HOXA11, COL3A1, COL1A1, MMP2, MMP9, TIMP, dan p53 pada penderita prolaps uteri dibandingkan pada perempuan tanpa prolaps uteri, serta mengetahui faktor risiko yang berhubungan dengan prolaps uteri.Studi potong lintang ini melibatkan 22 pasien prolaps uteri dan 22 tanpa prolaps uteri mulai Juni 2016 sampai Februari 2017 di RSUP dr. Kariadi Semarang. Dilakukan pencatatan data karakteristik berupa usia, paritas, IMT dan berat lahir bayi. Dilakukan pemeriksaan sekuens DNA gen HOXA11 dan Col3A1, pemeriksaan imunohistokimia pada ligamentum sakrouterina untuk menilai ekspresi protein HOXA11, COL1A1, Col3A1, MMP2, MMP9, TIMP, dan p53 pada perempuan menopause dengan prolaps uteri dan tanpa prolaps uteri.Tidak didapatkan variasi berupa mutasi gen HOXA11 pada perempuan dengan prolaps uteri sepanjang fragmen yang digunakan untuk sekuensing DNA. Didapatkan mutasi pada gen COL3A1 pada 10 pasien dengan prolaps uteri dan 6 pasien tanpa prolaps uteri p = 0,719 sepanjang fragmen yang digunakan untuk sekuensing DNA. Ekspresi protein COL1A1, MMP-9 dan p53 lebih tinggi pada kelompok prolaps p < 0,05 . Rerata usia, rerata paritas dan rerata berat lahir bayi, berbeda secara uji statistik pada kedua kelompok.Pada fragmen yang diperiksa tidak didapatkan mutasi gen HOXA11, namun didapatkan mutasi gen COL3A1 pada penderita prolaps maupun perempuan tanpa prolaps uteri. Tampak adanya faktor internal yang berperan untuk terjadinya prolaps uteri selain berbagai faktor risiko klinis. Faktor eksternal berupa usia, berat bayi lahir, dan paritas memiliki hubungan dengan prolaps uteri. Kata kunci: COL1A1, COL3A1, faktor klinis, HOXA11, menopause, MMP2, MMP9, mutasi gen, p53, prolaps uteri, TIMP.

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ABSTRACT
Uterine prolapse is a condition that decreases the quality of life of women. Multiple factors, mostly clinical, affect the course of uterine prolapse.The aims of the study were to investigate the genetic variation in the form of HOXA11 and Col3a2 gene mutations in women with uterine prolapse. This study also aimed to investigate different expression of HOXA11, COL3A1, COL1A1, MMP2, MMP9, TIMP, and p53 proteins in women with and without uterine prolapse, and to understand risk factors associated with uterine prolapse.A total of 44 women were enrolled in this cross sectional study, 22 of which with uterine prolapse and 22 others without uterine prolapse. This study was conducted between June 2016 and February 2017 in RSUP dr Kariadi, Semarang. demographic data including age, parity, BMI, and birth weight were recorded. HOXA11 and COL3A1 gene sequencing, immunohistochemistry testing of uterosacral ligament were conducted to assess HOXA11, COL1A1, COL3A1, MMP2, MMP9, TIMP, and p53 protein expressions.A mutation in COL3A1 gene along the fragment used in DNA sequencing was found among 10 women with uterine prolapse and 6 women without uterine prolapse although this did not reach statistical significance p .719 . No genetic variation in the form of HOXA11 gene mutation was found in women with uterine prolapse. Higher expression of COL1A1, MMP 9 and p53 proteins were found in prolapse group p .05 . The average of age, parity, and birth weight in two groups were statistically different.Only the COL3A1 gene mutation was detected in women with and without uterine prolapse. Beside, various clinical factors, it was confirmed that some internal factors also play important role in the course of uterine prolapse. Examples of external factors are age, birth weight, and parity. Key word COL1A1, COL3A1, clinical factors, gen mutation, HOXA11, menopause, MMP2, MMP9, p53, TIMP, uterine prolapse.