101
Lampiran 1. Perlakuan Mencit
Perlakuan Mencit
1. Mencit BALB/C yang digunakan adalah mencit BALB/c betina
berumur 6-8 minggu dengan berat 20-30 gram. Rambut di area
punggung dicukur sepanjang 2x3 cm, kemudian diolesi krim dasar dan
krim ekstrak daun kelengkeng dengan masing-masing konsentrasi
3,25%, 6,5%, 13% sehari 2 kali, seminggu 5 kali selama 4 minggu.
2. Kualitas-kuantitas kandang adalah kandang pemeliharaan dengan atap
dari kawat, dilengkapi dengan tempat makanan-minuman dan
disediakan satu kandang untuk tiap kelompok perlakuan yang berbeda
tiap mencit, yaitu tiap kandang berisi 6 mencit. Kualitas-kuantitas
makanan berupa konsentrat makanan ayam 30%, jagung giling 40% dan
dedak 30%, diberikan secara ad libitum. Minuman yang diberikan
secara tidak terbatas (ad libitum). Suhu ruangan dipertahankan 20-
25˚C. Kelembaban dan pertukaran udara yang ekstrim harus
dihindari. Aliran udara dalam ruangan harus lemah dan mantap (ruang
berventilasi baik dengan penyinaran normal).
102
Lampiran 2. Pewarnaan Kolagen dengan Teknik Mansson’s Trichome
Bahan yang dibutuhkan:
Fiksasi: formal saline
Solution A:
Acid fuchsin 0,5 g
Glacial acetic acid 0,5 g
Air detilasi 100 ml
Solution B:
Phosphomolybdic acid 1 g
Air destilasi 100 ml
Solution C
Methyl blue 2 g
Glacial acetic acid 2,5 g
Air destilasi 100 ml
Langkah:
a. Dilakukan deparafinisasi dan rehidrasi meliputi perendaman dalam alkohol
100%, 95% alkohol, 70% alkohol
b. Membersihkan pigmen merkuri dengan iodin, sodium thiosulfate
c. Cuci dengan air mengalir
d. Warnai nukleus (inti sel) dengan celestine blue hematoxyllin
e. Beri asam alkohol 1%
f. Cuci dengan air mengalir
g. Warnai dengan solution A asam fuchsin selama 5 menit
h. Cuci dengan air destilasi
i. Beri solution B asam phosphomolybdic seama 5 menit
j. Cuci
k. Warnai dengan solution C methyl blue selama 2-5 menit
l. Cuci dengan air destilasi
m. Beri asam acetic 1% selama 2 menit
n. Dehidrasi asending alkohol
o. Beri xylene lalu mount in permanent mounting pada medium berbasis
xylene (DPX).
Hasil Pengecatan:
Kolagen hijau/biru kehijauan
Nukleus biru/hitam
Sitoplasma, otot, eritrosit merah
103
Lampiran 3. Ethical Clearance
104
Lampiran 4. Surat Keterangan Penelitian di Laboratorium Farmakologi
105
Lampiran 5 Surat Keterangan Penelitian di Laboratorium Biologi
106
Lampiran 6. Surat Keterangan Penelitian di Laboratorium Kimia
107
Lampiran 7. Surat Keterangan Penelitian di Laboratorium Patologi Anatomi
108
Lampiran 8. Surat Keterangan Penelitian di Laboratorium Biologi UNNES
109
Lampiran 9. Tanaman Kelengkeng dan Krim Ekstrak Daun Kelengkeng
TUMBUHAN KELENGKENG
Krim Dasar dan Krim Ekstrak Daun Kelengkeng
110
Lampiran 10. Perlakuan Hewan Coba Mencit BALB/c
Mencit BALB/c dalam
kandang yang dibagi
K, P1, P2, P3
Mencit BALB/c
sebelum perlakuan
Mencit BALB/c
diolesi Krim ekstrak
daun kelengkeng
13%
Mencit BALB/c
diolesi Krim
ekstrak daun
kelengkeng 6,5%
Mencit BALB/c
diolesi Krim ekstrak
daun kelengkeng
3,25%
Mencit BALB/c diolesi
krim dasar
111
Lampiran 11. Lampiran hasil analisis deskriptif, normalitas data dan homogenitas
varian
Case Processing Summary
kelompok
Cases
Valid Missing Total
N Percent N Percent N Percent
Densita
Kolagen
kontrol 6 100.0% 0 0.0% 6 100.0%
perlakuan 1 6 100.0% 0 0.0% 6 100.0%
perlakuan 2 6 100.0% 0 0.0% 6 100.0%
perlakuan 3 6 100.0% 0 0.0% 6 100.0%
MMP-1 kontrol 6 100.0% 0 0.0% 6 100.0%
perlakuan 1 6 100.0% 0 0.0% 6 100.0%
perlakuan 2 6 100.0% 0 0.0% 6 100.0%
perlakuan 3 6 100.0% 0 0.0% 6 100.0%
fibroblas kontrol 6 100.0% 0 0.0% 6 100.0%
perlakuan 1 6 100.0% 0 0.0% 6 100.0%
perlakuan 2 6 100.0% 0 0.0% 6 100.0%
perlakuan 3 6 100.0% 0 0.0% 6 100.0%
Descriptives
Kelompok Statistic Std. Error
Densitas
Kolagen
kontrol Mean 1.00 .258
95% Confidence Interval
for Mean
Lower Bound .34
Upper Bound 1.66
5% Trimmed Mean 1.00
112
Median 1.00
Variance .400
Std. Deviation .632
Minimum 0
Maximum 2
Range 2
Interquartile Range 1
Skewness .000 .845
Kurtosis 2.500 1.741
perlakuan 1 Mean 1.33 .333
95% Confidence Interval
for Mean
Lower Bound .48
Upper Bound 2.19
5% Trimmed Mean 1.37
Median 1.50
Variance .667
Std. Deviation .816
Minimum 0
Maximum 2
Range 2
Interquartile Range 1
Skewness -.857 .845
Kurtosis -.300 1.741
perlakuan 2 Mean 1.50 .224
95% Confidence Interval Lower Bound .93
113
for Mean Upper Bound 2.07
5% Trimmed Mean 1.50
Median 1.50
Variance .300
Std. Deviation .548
Minimum 1
Maximum 2
Range 1
Interquartile Range 1
Skewness .000 .845
Kurtosis -3.333 1.741
perlakuan 3 Mean 2.00 .258
95% Confidence Interval
for Mean
Lower Bound 1.34
Upper Bound 2.66
5% Trimmed Mean 2.00
Median 2.00
Variance .400
Std. Deviation .632
Minimum 1
Maximum 3
Range 2
Interquartile Range 1
Skewness .000 .845
Kurtosis 2.500 1.741
114
MMP-1 kontrol Mean 3.533 .7548
95% Confidence Interval
for Mean
Lower Bound 1.593
Upper Bound 5.474
5% Trimmed Mean 3.404
Median 2.900
Variance 3.419
Std. Deviation 1.8490
Minimum 2.2
Maximum 7.2
Range 5.0
Interquartile Range 1.8
Skewness 2.148 .845
Kurtosis 4.857 1.741
perlakuan 1 Mean 2.733 .6687
95% Confidence Interval
for Mean
Lower Bound 1.014
Upper Bound 4.452
5% Trimmed Mean 2.737
Median 2.900
Variance 2.683
Std. Deviation 1.6379
Minimum 1.0
Maximum 4.4
Range 3.4
Interquartile Range 3.3
Skewness -.096 .845
115
Kurtosis -2.969 1.741
perlakuan 2 Mean 2.400 .5033
95% Confidence Interval
for Mean
Lower Bound 1.106
Upper Bound 3.694
5% Trimmed Mean 2.422
Median 2.400
Variance 1.520
Std. Deviation 1.2329
Minimum .6
Maximum 3.8
Range 3.2
Interquartile Range 2.3
Skewness -.300 .845
Kurtosis -1.138 1.741
perlakuan 3 Mean 1.333 .1333
95% Confidence Interval
for Mean
Lower Bound .991
Upper Bound 1.676
5% Trimmed Mean 1.326
Median 1.300
Variance .107
Std. Deviation .3266
Minimum 1.0
Maximum 1.8
Range .8
Interquartile Range .7
116
Skewness .383 .845
Kurtosis -1.481 1.741
fibroblas kontrol Mean 5.400 .5586
95% Confidence Interval
for Mean
Lower Bound 3.964
Upper Bound 6.836
5% Trimmed Mean 5.367
Median 4.800
Variance 1.872
Std. Deviation 1.3682
Minimum 4.0
Maximum 7.4
Range 3.4
Interquartile Range 2.5
Skewness .810 .845
Kurtosis -1.351 1.741
perlakuan 1 Mean 7.067 .7911
95% Confidence Interval
for Mean
Lower Bound 5.033
Upper Bound 9.100
5% Trimmed Mean 7.074
Median 6.700
Variance 3.755
Std. Deviation 1.9377
Minimum 4.2
Maximum 9.8
117
Range 5.6
Interquartile Range 3.1
Skewness .002 .845
Kurtosis .162 1.741
perlakuan 2 Mean 14.067 1.9695
95% Confidence Interval
for Mean
Lower Bound 9.004
Upper Bound 19.130
5% Trimmed Mean 13.841
Median 12.600
Variance 23.275
Std. Deviation 4.8244
Minimum 9.2
Maximum 23.0
Range 13.8
Interquartile Range 6.3
Skewness 1.538 .845
Kurtosis 2.781 1.741
perlakuan 3 Mean 24.500 2.8241
95% Confidence Interval
for Mean
Lower Bound 17.241
Upper Bound 31.759
5% Trimmed Mean 24.433
Median 22.500
Variance 47.852
Std. Deviation 6.9175
118
Minimum 16.0
Maximum 34.2
Range 18.2
Interquartile Range 12.8
Skewness .489 .845
Kurtosis -1.155 1.741
Tests of Normality
kelompok
Kolmogorov-Smirnova Shapiro-Wilk
Statistic df Sig. Statistic df Sig.
Densitas Kolagen
kontrol .333 6 .036 .827 6 .101
perlakuan 1 .293 6 .117 .822 6 .091
perlakuan 2 .319 6 .056 .683 6 .004
perlakuan 3 .333 6 .036 .827 6 .101
MMP-1 kontrol .362 6 .014 .714 6 .009
perlakuan 1 .280 6 .153 .801 6 .060
perlakuan 2 .168 6 .200* .952 6 .755
perlakuan 3 .180 6 .200* .920 6 .505
fibroblas kontrol .282 6 .148 .861 6 .193
perlakuan 1 .221 6 .200* .956 6 .786
perlakuan 2 .238 6 .200* .867 6 .215
perlakuan 3 .253 6 .200* .925 6 .543
*. This is a lower bound of the true significance.
a. Lilliefors Significance Correction
119
Test of Homogeneity of Variance
Levene Statistic df1 df2 Sig.
Densitas Kolagen
Based on Mean .917 3 20 .451
Based on Median .873 3 20 .472
Based on Median and with adjusted df
.873 3 14.412 .478
Based on trimmed mean .914 3 20 .452
MMP-1 Based on Mean 3.111 3 20 .049
Based on Median 1.992 3 20 .148
Based on Median and with adjusted df
1.992 3 6.930 .205
Based on trimmed mean 2.638 3 20 .078
fibroblas Based on Mean 4.551 3 20 .014
Based on Median 2.115 3 20 .130
Based on Median and with adjusted df
2.115 3 11.021 .156
Based on trimmed mean 4.297 3 20 .017
120
Lampiran 12 Hasil analisis perbedaan Densitas Kolagen, MMP-1 dan fibroblas antar 4
kelompok
NPar Tests Kruskal-Wallis Test
Ranks
kelompok N Mean Rank
Densitas Kolagen Kontrol 6 8.25
perlakuan 1 6 11.75
perlakuan 2 6 12.75
perlakuan 3 6 17.25
Total 24
MMP-1 Kontrol 6 16.83
perlakuan 1 6 13.92
perlakuan 2 6 12.75
perlakuan 3 6 6.50
Total 24
fibroblas Kontrol 6 5.25
perlakuan 1 6 7.92
perlakuan 2 6 15.92
perlakuan 3 6 20.92
Total 24
Test Statisticsa,b
trichom manson MMP-1 fibroblas
Chi-Square 5.948 6.860 18.754
df 3 3 3
Asymp. Sig. .114 .076 .000
a. Kruskal Wallis Test
b. Grouping Variable: kelompok
121
Lampiran 13. Uji korelasi Densitas Kolagen dan MMP-1
Hasil uji deskriptif dan normalitas data densitas kolagen dengan ekspresi MMP-1
dan jumlah fibroblas
Explore
Case Processing Summary
Cases
Valid Missing Total
N Percent N Percent N Percent
densitas kolagen 24 100.0% 0 0.0% 24 100.0%
MMP-1 24 100.0% 0 0.0% 24 100.0%
fibroblas 24 100.0% 0 0.0% 24 100.0%
Descriptives
Statistic Std. Error
densitas kolagen Mean 1.46 .147
95% Confidence
Interval for Mean
Lower
Bound 1.15
Upper Bound 1.76
5% Trimmed Mean 1.46
Median 1.50
Variance .520
Std. Deviation .721
Minimum 0
Maximum 3
122
Descriptives
Statistic Std. Error
Range 3
Interquartile Range 1
Skewness -.222 .472
Kurtosis -.076 .918
MMP-1 Mean 2.500 .3116
95% Confidence
Interval for Mean
Lower
Bound 1.855
Upper Bound 3.145
5% Trimmed Mean 2.367
Median 2.100
Variance 2.330
Std. Deviation 1.5266
Minimum .6
Maximum 7.2
Range 6.6
Interquartile Range 2.3
Skewness 1.299 .472
Kurtosis 2.378 .918
fibroblas Mean 12.758 1.7757
95% Confidence
Interval for Mean
Lower
Bound 9.085
Upper Bound 16.432
123
Descriptives
Statistic Std. Error
5% Trimmed Mean 12.078
Median 9.500
Variance 75.673
Std. Deviation 8.6990
Minimum 4.0
Maximum 34.2
Range 30.2
Interquartile Range 12.8
Skewness 1.132 .472
Kurtosis .467 .918
Tests of Normality
Kolmogorov-Smirnova Shapiro-Wilk
Statistic df Sig. Statistic df Sig.
densitas kolagen .274 24 .000 .839 24 .001
MMP-1 .135 24 .200* .892 24 .015
fibroblas .175 24 .056 .861 24 .004
*. This is a lower bound of the true significance.
a. Lilliefors Significance Correction