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Lampiran 1. Data Hasil Pemeriksaan Cairan Bronchoalveolar
Produksi NO makrofag
Index Produksi ROI makrofag
Kontrol
136.8
3.1
Kontrol
120.9
2.5
Kontrol
142.4
2.6
Kontrol
96.2
3
Kontrol
146.9
2.6
P1
106.1
2.5
P1
110.9
2.6
P1
141.5
2.5
P1
126.7
2.8
P1
138
2.7
P2
148.1
2.9
P2
156.7
2.8
P2
127.1
2.9
P2
148.4
2.8
P2
130.9
3.1
P3
160.8
2.8
P3
160.8
2.6
P3
136.7
2.8
P3
142.9
2.9
P3
143.8
2.5
Kelompok
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Lampiran 2. Foto Produksi NO Makrofag Bronchoalveolar
Foto Produksi ROI Makrofag Bronchoalveolar
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Lampiran 3. Hasil Analisa Statistik
a. Nitric Oxide
Cas e Proces s ing Sum m ary Cases Missing N Percent 0 ,0%
Valid N Nitric oxide
20
Percent 100,0%
Total N
Percent 100,0%
20
Des criptives Nitric oxide
Mean 95% Conf idence Interval f or Mean
Low er Bound Upper Bound
5% Trimmed Mean Median Varianc e Std. Deviation Minimum Max imum Range Interquartile Range Skew nes s Kurtosis
Statistic 136,130 127,960
Std. Error 3,9034
144,300 136,978 139,750 304,725 17,4564 96,2 160,8 64,6 21,0 -,740 ,206
,512 ,992
Tes ts of Nor mality a
Nitric oxide
Kolmogorov-Smirnov Statistic df Sig. ,163 20 ,171
a. Lilliefors Significance Correction
Shapiro-Wilk Statistic df ,943 20
Sig. ,278
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Kontrol Statistics Nitric oxide N Percentiles
Valid Mis sing 25 50 75
5 0 108,550 136,800 144,650
Perlakuan 1
Statistics Nitric oxide N Percentiles
Valid Mis sing 25 50 75
5 0 108,500 126,700 139,750
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Perlakuan 2 Statistics Nitric oxide N Percentiles
Valid Mis sing 25 50 75
5 0 129,000 148,100 152,550
Perlakuan 3
Statistics Nitric oxide N Percentiles
Valid Mis sing 25 50 75
5 0 139,800 143,800 160,800
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Des crip tives Nitric oxide
Perlakuan 0
1
2
3
Mean 95% Conf idence Interval f or Mean 5% Trimmed Mean Median V arianc e Std. Deviation Minimum Max imum Range Interquartile Range Skew nes s Kurtosis Mean 95% Conf idence Interval f or Mean 5% Trimmed Mean Median V arianc e Std. Deviation Minimum Max imum Range Interquartile Range Skew nes s Kurtosis Mean 95% Conf idence Interval f or Mean 5% Trimmed Mean Median V arianc e Std. Deviation Minimum Max imum Range Interquartile Range Skew nes s Kurtosis Mean 95% Conf idence Interval f or Mean 5% Trimmed Mean Median V arianc e Std. Deviation Minimum Max imum Range Interquartile Range Skew nes s Kurtosis
Low er Bound Upper Bound
Low er Bound Upper Bound
Low er Bound Upper Bound
Low er Bound Upper Bound
Statistic 128,640 103,030
Std. Error 9,2239
154,250 129,428 136,800 425,403 20,6253 96,2 146,9 50,7 36,1 -1,205 ,720 124,640 105,012
,913 2,000 7,0693
144,268 124,733 126,700 249,878 15,8075 106,1 141,5 35,4 31,3 -,188 -2,665 142,240 126,544
,913 2,000 5,6533
157,936 142,278 148,100 159,798 12,6411 127,1 156,7 29,6 23,6 -,303 -2,350 149,000 135,201
,913 2,000 4,9700
162,799 149,028 143,800 123,505 11,1133 136,7 160,8 24,1 21,0 ,321 -2,773
,913 2,000
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Oneway Des criptives Nitric oxide
N 0 1 2 3 Total
5 5 5 5 20
Mean 128,640 124,640 142,240 149,000 136,130
Std. Deviation 20,6253 15,8075 12,6411 11,1133 17,4564
Std. Error 9,2239 7,0693 5,6533 4,9700 3,9034
95% Confidence Interval for Mean Low er Bound Upper Bound 103,030 154,250 105,012 144,268 126,544 157,936 135,201 162,799 127,960 144,300
Minimum 96,2 106,1 127,1 136,7 96,2
Maximum 146,9 141,5 156,7 160,8 160,8
Tes t of Hom ogene ity of V ariance s Nitric oxide Levene Statistic ,975
df 1
df 2 3
Sig. ,429
16
ANOV A Nitric oxide
Betw een Groups Within Groups Total
Sum of Squares 1955,446 3834,336 5789,782
df 3 16 19
Mean Square 651,815 239,646
F 2,720
Sig. ,079
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b. Reactive Oxygen Intermediate Cas e Proce ss ing Sum m ary
N Reac tive Ox igen Intermediate
Cases Mis sing N Percent
Valid Percent 20
100.0%
0
.0%
N
Total Percent 20
100.0%
Des criptives Reac tive Ox igen Intermediate
Mean 95% Conf idence Interval f or Mean
Statistic 2,750 2,658
Low er Bound Upper Bound
Std. Error ,0438
2,842
5% Trimmed Mean Median Varianc e Std. Deviation Minimum Max imum Range Interquartile Range Skew nes s Kurtosis
2,744 2,800 ,038 ,1960 2,5 3,1 ,6 ,3 ,280 -,939
,512 ,992
Tes ts of Normality a
Kolmogorov-Smirnov Statistic df Sig. Reactive Oxigen Intermediate
,178
a. Lilliefors Significance Correction
20
,097
Shapiro-Wilk Statistic df ,916
20
Sig. ,083
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Kontrol Statistics Reac tive Ox igen Intermediate N Valid 5 Mis sing 0 Percentiles 25 2,550 50 2,600 75 3,050
Perlakuan 1 Statistics Reac tive Ox igen Intermediate N V alid 5 Mis sing 0 Percentiles 25 2,500 50 2,600 75 2,750
+
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Perlakuan 2 Statistics Reac tive Ox igen Intermediate N Valid 5 Mis sing 0 Percentiles 25 2,800 50 2,900 75 3,000
Perlakuan 3 Statistics Reac tive Ox igen Intermediate N Valid 5 Mis sing 0 Percentiles 25 2,550 50 2,800 75 2,850
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Des criptives Reac tive Ox igen Intermediate
Perlakuan 0
1
2
3
Mean 95% Conf idence Interval f or Mean 5% Trimmed Mean Median V arianc e Std. Deviation Minimum Max imum Range Interquartile Range Skew nes s Kurtosis Mean 95% Conf idence Interval f or Mean 5% Trimmed Mean Median V arianc e Std. Deviation Minimum Max imum Range Interquartile Range Skew nes s Kurtosis Mean 95% Conf idence Interval f or Mean 5% Trimmed Mean Median V arianc e Std. Deviation Minimum Max imum Range Interquartile Range Skew nes s Kurtosis Mean 95% Conf idence Interval f or Mean 5% Trimmed Mean Median V arianc e Std. Deviation Minimum Max imum Range Interquartile Range Skew nes s Kurtosis
Low er Bound Upper Bound
Low er Bound Upper Bound
Low er Bound Upper Bound
Low er Bound Upper Bound
Statistic 2,760 2,425
Std. Error ,1208
3,095 2,756 2,600 ,073 ,2702 2,5 3,1 ,6 ,5 ,578 -2,708 2,620 2,458
,913 2,000 ,0583
2,782 2,617 2,600 ,017 ,1304 2,5 2,8 ,3 ,3 ,541 -1,488 2,900 2,748
,913 2,000 ,0548
3,052 2,894 2,900 ,015 ,1225 2,8 3,1 ,3 ,2 1,361 2,000 2,720 2,516
,913 2,000 ,0735
2,924 2,722 2,800 ,027 ,1643 2,5 2,9 ,4 ,3 -,518 -1,687
,913 2,000
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Oneway Des criptives Reactive Oxigen Intermediate
N 0 1 2 3 Total
5 5 5 5 20
Mean Std. Deviation Std. Error 2,760 ,2702 ,1208 2,620 ,1304 ,0583 2,900 ,1225 ,0548 2,720 ,1643 ,0735 2,750 ,1960 ,0438
95% Confidence Interval for Mean Low er Bound Upper Bound 2,425 3,095 2,458 2,782 2,748 3,052 2,516 2,924 2,658 2,842
Tes t of Hom ogene ity of V ariance s Reac tive Ox igen Intermediate Levene Statistic 4,436
df 1
df 2 3
16
Sig. ,019
Kruskal-Wallis Test
Ranks Reac tive Ox igen Intermediate
Perlakuan 0 1 2 3 Total
N
a,b T es t Statis tics
Chi-Square df A sy mp. Sig.
Reac tive Oxigen Intermediate 5,568 3 ,135
a. Kruskal Wallis Test b. Grouping V ariable: Perlakuan
5 5 5 5 20
Mean Rank 10,60 6,50 15,10 9,80
Minimum 2,5 2,5 2,8 2,5 2,5
Maximum 3,1 2,8 3,1 2,9 3,1
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Lampiran 4. Cara Pemeliharaan, Pemberian Jus Mengkudu, Dekapitasi dan Pengambilan Cairan Bronchoalveolar a. Cara pemeliharaan Tikus wistar yang dipilih berumur sekitar 15 minggu dengan berat badan rata-rata 200 gram, dipelihara pada kandang selama 30 hari. Masing-masing kandang diisi 5 ekor tikus. Tikus diberi makan dan minum yang sama. b. Cara memegang tikus dan pemberian jus mengkudu Pengambilan tikus dari kandang dilakukan dengan mengambil ekornya, kemudian tikus diletakkan di atas kasa, selanjutnya bagian punggung tikus dipegang dengan telapak tangan dengan jari-jari memegang bagian leher sehingga kepala tikus dalam posisi terangkat ke atas untuk pemberian jus mengkudu melalui sonde. c. Cara dekapitasi dan pengambilan cairan bronchoalveolar 1) Tikus wistar diterminasi dengan dekapitasi leher, dengan cara meletakkan tikus di atas permukaan rata. Sebuah pinset diletakkan di atas kuduk tikus. Sambil menekan pinset, ekor tikus ditarik kuat dan pinset diarahkan ke atas kepala tikus. 2) Tikus kemudian diletakkan pada posisi terlentang dan seluruh permukaan leher sampai ventral disiram dengan alkohol 70%. 3) Dibuat irisan kecil melalui sayatan ventral garis tengah sekitar 5 cm panjang di leher. Kulit dirobek dengan menggunakan pinset hingga kulit terkelupas, dan tampak rongga pleura.
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4) Trakhea yang terlihat dihubungkan dengan kanula dengan tabung polietilen yang terhubung dengan jarum suntik dengan ukuran 20G, kemudian paru-paru dicuci dengan dimasukkan 3 ml larutan PBS, dilakukan beberapa kali hingga didapatkan cairan bronchoalveolar dengan jumlah yang cukup.
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Lampiran 5. Cara Pembuatan Reagen Griess dan Standard Nitrit
a. Cara membuat reagen Griess: Reagen1: N-(1-naphthyl) ethylenediamine dihydrochloride = NED (Sigma): 0,1 gram dilarutkan dalam 100 ml air suling. Reagen 2: Sulfanilamide (Sigma): 1 gram dilarutkan dalam 100 ml HCl 4 N. Reagen 1 dan reagen 2 harus disimpan dalam almari pendingin dalam botol gelap dan dapat digunakan dalam 6 minggu atau selama tidak berubah warna menjadi lebih gelap. Reagen Griess: mencampur dengan volume sama banyak antara reagen 1 dan reagen 2 setiap akan digunakan. Reagen siap digunakan dalam 1 jam setelah pencampuran. b. Cara membuat Standard Nitrit : 69 mg NaNO2 dilarutkan dalam 500 ml air suling ( 2mM stok), kemudian dibuat pengenceran bertingkat dari 0 – 200 uM dengan cara melarutkan larutan stok menggunakan medium yang dipakai untuk kultur makrofag.
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66
