BAB VII SIMPULAN DAN SARAN
7.2 Saran
Sebagai saran dalam penelitian ini adalah:
1. Perlu melakukan penelitian lebih lanjut pada tikus Wistar untuk mengetahui efektifitas pemberian ekstrak pegagan oral terhadap peningkatan jumlah kolagen dan penurunan ekspresi MMP-1.
2. Perlu melakukan penelitian klinis (uji klinis) pada manusia untuk mengetahui efektifitas pemberian ekstrak pegagan oral terhadap peningkatan jumlah kolagen dan penurunan ekspresi MMP-1 pada peremajaan kulit dan menghambat penuaan kulit.
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Lampiran 1
Lampiran-2 Efek UV terhadap kulit
UV-C UV-B UV-A
Wavelength: 100-280 nm Higher energy per photon. Wavelength: 280-315 nm Intermediate energy per photon. Wavelength: 315-400 nm Lower energy per photon Sources: • Sun (UV-C is absorbed by molecular oxygen, ozone and water vapour in the upper atmosphere) • Germicidal lamps • Arcwelding equipment • High intensity discharge lamps (HIDL) Sources: • Sun (5% of UVR at ground level, only wavelengths > 297 nm) • Germicidal lamps • Arc welding equipment • HIDL • Therapeutics lamps • Medical and industrial lasers Sources: • Sun (95% of UVR at ground level)
• Black light lamps
• Germicidal lamps • Arc welding equipment • HIDL • Therapeutics lamps • Tanning devices (sunbeds) Penetration: • Photons between 100 to 200 nm are absorbed in air. • Absorbed by keratin in the epidermis, does not penetrate to the dermis. Penetration: • Partially absorbed by ozone in the upper atmosphere • Penetrates to the dermis Penetration : • Not absorbed by ozone • Penetrates deeper into the skin than any other form of UVR Effects: • DNA damage Effects: • Responsible Effects: • Causes cxxi
on unprotected cells: epithelium, cornea and bacteria. for vitamin D3 production and delayed tanning. • Most effective in causing acute and chronic harmful effects. • Sunburn, immunosuppr ession, cellular damage, skin cancer, solar urticaria, photo aging and, photokerato-conjunctivitis, cataract, and pterygium. immediate tanning. • Can potentiate some carcinogeni c effects of UVB. • Thermal burns • Sunburn, immuno- suppression , cellular damage, photoallerg y, phototoxicit y, photo-aging, photokerat o-conjunctivit is, cataract and pterygium, solar retinitis. cxxii
Lampiran 3. Penanganan Hewan Coba
Pengelolaan Hewan Coba pada penelitian dengan judul:
PEMBERIAN ORAL EKSTRAK DAUN PEGAGAN (Centella asiatica) LEBIH BANYAK MENINGKATKAN JUMLAH KOLAGEN DAN MENURUNKAN EKSPRESI MMP-1 DARIPADA VITAMIN C PADA TIKUS WISTAR (Rattus norvegicus) YANG DIPAPAR SINAR UV-B
Sesuai dengan saran dari Komisi etik Penelitian FK UNUD maka hewan
coba yang dipilih sebagai sampel diperlakukan dengan baik agar kenyamanan
hewan yang telah berkorban untuk kepentingan kemanusiaan tetap terjamin.
Perlakuan sebelum penelitian:
Tikus yang akan dipilih sebagai sampel harus homogen berdasarkan umur
dan berat badannya. Tikus yang dipakai didapat dari Laboratorium Farmakologi
FK UNUD dan dipelihara dalam kandang yang dibuat nyaman. Ukuran kandang
tikus adalah 60 X 40 X 60 cm, dengan kebersihan, sirkulasi udara, penerangan
dan penyediaan makan dan minum yang terjamin selama 24 jam. Setiap kandang
diberi alas tidur dengan sekam agar mampu menghisap air kemih dan agar
kandang tetap kering serta tidak mengandung zat kimia, setiap kandang ditempati
oleh 3 ekor tikus.
Sebelum mulai penelitian, bulu pada semua tikus pada bagian
punggungnya dicukur dengan alat pencukur rambut dan skapel dengan ukuran 5 x
5 cm.
Perlakuan selama penelitian:
Selama penelitian tikus diletakkan secara teratur dengan nomor urut sesuai
kelompok. Makanan dan air minum dimonitor sehari 2 kali (pagi dan sore), suhu
dan ventilasi serta kelembaban kandang dijaga dengan baik.
Semua tikus Wistar dari semua kelompok yang telah dicukur bulu
punggungnya diberikan paparan sinar UVB sebanyak 3 kali seminggu. Kemudian
masing-masing kelompok perlakuan diberikan obat oral sesuai dosis pada masing
masing kelompok perlakuan. Perlakuan pertama diberikan aquadest 1 ml (kontrol
dan dipapar sinar UV-B, perlakuan kedua ekstrak pegagan 50 mg/200 mgBB tikus
secara oral diberikan sekali sehari dan dipapar sinar UV-B, perlakuan ketiga
diberikan vitamin C dengan dosis 9 mg/200 mgBB tikus sekali sehari dan dipapar
sinar UV-B. Setelah 48 jam dari penyinaran terakhir, semua tikus Wistar dari
ketiga kelompok perlakuan dilakukan eutanasia dengan kloroform kemudian
dilakukan biopsi jaringan kulit dengan cara diambil jaringan kulitnya dengan
ukuran 1 cm x 1 cm dan dibagi menjadi 3 bagian yaitu untuk pengukuran jumlah
kolagen dan ekspresi MMP-1.
Perlakuan sesudah penelitian:
Tikus yang sudah didekapitasi kemudian dikubur dengan baik.
Lampiran 4
Lampiran 5
Lampiran 6
Lampiran 7
Lampiran 8
Tests of Normality
Kelompok
Kolmogorov-Smirnova Shapiro-Wilk
Statistic df Sig. Statistic df Sig.
Kolagen Kontrol .309 6 .076 .894 6 .340 Perlakuan 1 .175 6 .200* .968 6 .881 Perlakuan 2 .241 6 .200* .889 6 .315 Perlakuan 3 .243 6 .200* .908 6 .423 Vitamin C .297 6 .105 .826 6 .100 MMP_1 Kontrol .182 6 .200* .962 6 .838 Perlakuan 1 .244 6 .200* .923 6 .528 Perlakuan 2 .278 6 .161 .870 6 .224 Perlakuan 3 .282 6 .148 .838 6 .125 Vitamin C .174 6 .200* .968 6 .878
a. Lilliefors Significance Correction
*. This is a lower bound of the true significance.
Test of Homogeneity of Variances
Levene Statistic df1 df2 Sig.
Kolagen 1.345 4 25 .281
MMP_1 1.666 4 25 .189
Lampiran 9 Oneway Descriptives N Mean Std. Deviatio n Std. Error 95% Confidence Interval for Mean
Minimu m Maximu m Lower Bound Upper Bound Kolage n Kontrol 6 50.5133 3.61508 1.47585 46.7195 54.3071 44.12 55.11 Perlakuan 1 6 54.9033 2.44638 .99873 52.3360 57.4707 51.47 58.34 Perlakuan 2 6 60.1217 4.25127 1.73557 55.6602 64.5831 52.48 64.46 Perlakuan 3 6 59.3083 1.44576 .59023 57.7911 60.8256 56.93 60.74 Vitamin C 6 55.7217 1.29327 .52797 54.3645 57.0789 54.58 57.57 Total 30 56.1137 4.39420 .80227 54.4728 57.7545 44.12 64.46 MMP_ 1 Kontrol 6 29.0967 3.40441 1.38984 25.5240 32.6694 24.12 34.51 Perlakuan 1 6 14.6233 3.44003 1.40439 11.0132 18.2334 10.64 19.44 Perlakuan 2 6 11.0117 1.47471 .60205 9.4641 12.5593 8.33 12.50 Perlakuan 3 6 9.4650 1.67664 .68448 7.7055 11.2245 8.00 11.76 Vitamin C 6 14.0067 1.44120 .58837 12.4942 15.5191 12.10 16.34 Total 30 15.6407 7.47051 1.36392 12.8511 18.4302 8.00 34.51 ANOVA
Sum of Squares df Mean Square F Sig.
Kolagen Between Groups 355.514 4 88.878 10.868 .000
Within Groups 204.448 25 8.178 Total 559.962 29 MMP_1 Between Groups 1466.012 4 366.503 60.109 .000 Within Groups 152.434 25 6.097 Total 1618.446 29 cxxxii
Post Hoc Tests Multiple Comparisons LSD Dependent Variable (I) Kelompok (J) Kelompok Mean Difference
(I-J) Std. Error Sig.
95% Confidence Interval Lower
Bound
Upper Bound Kolagen Kontrol Perlakuan 1 -4.39000* 1.65105 .013 -7.7904 -.9896
Perlakuan 2 -9.60833* 1.65105 .000 -13.0087 -6.2079 Perlakuan 3 -8.79500* 1.65105 .000 -12.1954 -5.3946 Vitamin C -5.20833* 1.65105 .004 -8.6087 -1.8079 Perlakuan 1 Kontrol 4.39000* 1.65105 .013 .9896 7.7904 Perlakuan 2 -5.21833* 1.65105 .004 -8.6187 -1.8179 Perlakuan 3 -4.40500* 1.65105 .013 -7.8054 -1.0046 Vitamin C -.81833 1.65105 .624 -4.2187 2.5821 Perlakuan 2 Kontrol 9.60833* 1.65105 .000 6.2079 13.0087 Perlakuan 1 5.21833* 1.65105 .004 1.8179 8.6187 Perlakuan 3 .81333 1.65105 .627 -2.5871 4.2137 Vitamin C 4.40000* 1.65105 .013 .9996 7.8004 Perlakuan 3 Kontrol 8.79500* 1.65105 .000 5.3946 12.1954 Perlakuan 1 4.40500* 1.65105 .013 1.0046 7.8054 Perlakuan 2 -.81333 1.65105 .627 -4.2137 2.5871 Vitamin C 3.58667* 1.65105 .040 .1863 6.9871 Vitamin C Kontrol 5.20833* 1.65105 .004 1.8079 8.6087 Perlakuan 1 .81833 1.65105 .624 -2.5821 4.2187 Perlakuan 2 -4.40000* 1.65105 .013 -7.8004 -.9996 Perlakuan 3 -3.58667* 1.65105 .040 -6.9871 -.1863 MMP_1 Kontrol Perlakuan 1 14.47333* 1.42564 .000 11.5372 17.4095 Perlakuan 2 18.08500* 1.42564 .000 15.1488 21.0212 Perlakuan 3 19.63167* 1.42564 .000 16.6955 22.5678 Vitamin C 15.09000* 1.42564 .000 12.1538 18.0262 Perlakuan 1 Kontrol -14.47333* 1.42564 .000 -17.4095 -11.5372 Perlakuan 2 3.61167* 1.42564 .018 .6755 6.5478 Perlakuan 3 5.15833* 1.42564 .001 2.2222 8.0945 Vitamin C .61667 1.42564 .669 -2.3195 3.5528 Perlakuan 2 Kontrol -18.08500* 1.42564 .000 -21.0212 -15.1488 Perlakuan 1 -3.61167* 1.42564 .018 -6.5478 -.6755 cxxxiii
Perlakuan 3 1.54667 1.42564 .288 -1.3895 4.4828 Vitamin C -2.99500* 1.42564 .046 -5.9312 -.0588 Perlakuan 3 Kontrol -19.63167* 1.42564 .000 -22.5678 -16.6955 Perlakuan 1 -5.15833* 1.42564 .001 -8.0945 -2.2222 Perlakuan 2 -1.54667 1.42564 .288 -4.4828 1.3895 Vitamin C -4.54167* 1.42564 .004 -7.4778 -1.6055 Vitamin C Kontrol -15.09000* 1.42564 .000 -18.0262 -12.1538 Perlakuan 1 -.61667 1.42564 .669 -3.5528 2.3195 Perlakuan 2 2.99500* 1.42564 .046 .0588 5.9312 Perlakuan 3 4.54167* 1.42564 .004 1.6055 7.4778 *. The mean difference is significant at the 0.05 level.
Lampiran 10. Foto Aktivitas Penelitian
Tikus Wistar Jantan
Tikus Wistar Jantan Proses Penimbangan tikus Wistar
Kelompok perlakuan tikus Wistar jantan dalam kandang
Pencukuran Bulu tikus Wistar Dekapitasi tikus Wistar
yang akan Diberi Perlakuan
tikus Wistar sedang dicukur tikus Wistar diambil jaringan kulitnya
Tikus Wistar sedang dipapar UVB dalam box Simulator UVB & box penyinaran tikus Wistar
Simulator UVB & box penyinaran tikus Wistar Penyinaran UV-B pada Kelompok tikus Wistar
Pengambilan Jaringan Kulit ukuran 1 x 1 cm2 Dekapitasi tikus Wistar Pada punggung tikus Wistar
untuk Pembuatan Preparat Histologi
Daun Pegagan Vacuum Rotary Evaporator
Isi kit LSAB+ DAKO
Kit LSAB+ DAKO, Fetal bovine Serum, dan anti bodi MMP1
Mikroskop Olympus CX41 Microtome Leica Jung 820 dan kamera Optilab pro