101 - repository.unissula.ac.idrepository.unissula.ac.id/13414/12/lampiran.pdfdisediakan satu...

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


for Mean

Lower Bound .48

Upper Bound 2.19


Median 1.50

Variance .667

Std. Deviation .816

Minimum 0

Maximum 2

Range 2


Skewness -.857 .845

Kurtosis -.300 1.741


95% Confidence Interval Lower Bound .93

113

for Mean Upper Bound 2.07


Median 1.50

Variance .300

Std. Deviation .548

Minimum 1

Maximum 2

Range 1


Skewness .000 .845

Kurtosis -3.333 1.741



for Mean

Lower Bound 1.34

Upper Bound 2.66


Median 2.00

Variance .400

Std. Deviation .632

Minimum 1

Maximum 3

Range 2


Skewness .000 .845


114

MMP-1 kontrol Mean 3.533 .7548


for Mean

Lower Bound 1.593

Upper Bound 5.474


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




for Mean

Lower Bound 1.014

Upper Bound 4.452


Median 2.900

Variance 2.683


Minimum 1.0

Maximum 4.4

Range 3.4


Skewness -.096 .845

115

Kurtosis -2.969 1.741



for Mean

Lower Bound 1.106

Upper Bound 3.694


Median 2.400

Variance 1.520


Minimum .6

Maximum 3.8

Range 3.2


Skewness -.300 .845

Kurtosis -1.138 1.741



for Mean

Lower Bound .991

Upper Bound 1.676


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


for Mean

Lower Bound 3.964

Upper Bound 6.836


Median 4.800

Variance 1.872


Minimum 4.0

Maximum 7.4

Range 3.4


Skewness .810 .845

Kurtosis -1.351 1.741



for Mean

Lower Bound 5.033

Upper Bound 9.100


Median 6.700

Variance 3.755


Minimum 4.2

Maximum 9.8

117

Range 5.6


Skewness .002 .845

Kurtosis .162 1.741

perlakuan 2 Mean 14.067 1.9695


for Mean

Lower Bound 9.004

Upper Bound 19.130


Median 12.600

Variance 23.275


Minimum 9.2

Maximum 23.0

Range 13.8


Skewness 1.538 .845


perlakuan 3 Mean 24.500 2.8241


for Mean

Lower Bound 17.241

Upper Bound 31.759


Median 22.500

Variance 47.852


118

Minimum 16.0

Maximum 34.2

Range 18.2


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


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


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


Median 1.50

Variance .520

Std. Deviation .721

Minimum 0

Maximum 3

122

Descriptives


Range 3


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


Median 2.100

Variance 2.330


Minimum .6

Maximum 7.2

Range 6.6


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



Median 9.500

Variance 75.673


Minimum 4.0

Maximum 34.2

Range 30.2


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

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