Documente Academic
Documente Profesional
Documente Cultură
5M
GRADUATING PAPER
Written by :
Muhammad
Herdhana Ash
Shidiqi
12/328510/KU/
14897
FACULTY OF MEDICINE
Universitas GADJAH MADA
YOGYAKARTA
2015
AUTHENTIC STATEMENT
I, the undersigned
1
Name : Muhammad Herdhana Ash Shidiqi
Faculty : Medicine
in footnotes.
The writers
APPROVAL PAGE
i
EXPRESSION OF LUTEINIZING HORMONE RECEPTOR (LHR) IN THE LEYDIG
CELL CULTURE OF SPRAGUE DAWLEY RAT THAT INDUCED BY ADVANCED
GLYCATION END-PRODUCT 200G/ML AND GAMMA MANGOSTEEN 5M
By :
2
Tested and approved on -
Expert Advisor
NIP 197007091999032001
ACKNOWLEDGEMENT
3
At the same time to my other friends, seniors and
secretaries at the international program, your help is deeply
appriciated for lending me a hand in completing my thesis.
Finally, i really hope that this thesis will be
beneficial for the readers and for the advanvement or medical
theory.
Thank you
4
Contents
AUTHENTIC STATEMENT....................................................................................................II
APPROVAL PAGE....................................................................................................................II
ACKNOWLEDGEMENT.......................................................................................................III
LIST OF FIGURES....................................................................................................................3
TABLE LIST..............................................................................................................................4
LIST OF ATTACHMENT..........................................................................................................5
ABBREVIATION LIST.............................................................................................................6
ABSTRACT...............................................................................................................................7
CHAPTER I. INTRODUCTION...............................................................................................9
I.1. Background....................................................................................................................10
I.2.Problem Formulation......................................................................................................13
I.3. Objective........................................................................................................................13
I.4. Research Authenticity....................................................................................................14
I.5. Research Benefit............................................................................................................14
LIST OF FIGURES
inflammatory cytokines 25
TABLE LIST
Characteristic 47
Table 2. The Mean Value of Post-Hoc Least Significant
Difference(LSD) analysis.
47
LIST OF ATTACHMENT
Attachment1. Detail of Data
Statisti 61
Normality of Data Distribution
Attachment2.
Tes
61
Attachment3. One way ANOVA Test and Test of
Homogeneity of
Variances 62
Attachment4. post hoc Statistical
Test 63
ABBREVIATION LIST
3-DG :3-deoxyglucosone
CML :Carboxymethyl-lysine
DHT :Dihidrotestosterone
LH :Luteinizing Hormone
PGE2 :Prostaglandin E2
INTISARI
Latar Belakang: Advanced Glycation End Products (AGE) adalah
protein atau lipid yang menjadi hasil dari serangkaian
reaksi kimia setelah awalan reaksi glikasi. AGE akan
menyebabkan seuatu kerusakan pada sel dengan peningkatan
stres oksidatif dan inflamasi. Gamma mangosteen merupakan
senyawa bioaktif utama pada extrak manggis dan merupakan
salah satu zat utama pada klasifikasi turunan xanthone.
Gamma mangostin mengandung antioksidan dan dapat menurunkan
respon inflamasi yang di sebabkan AGE.
CHAPTER I. INTRODUCTION
I.1. Background
free radicals. For this case also could be found in the people
(Bierhaus et al.1997).
found in Indonesia.
al., 2009).
I.2.Problem Formulation
formulated as below:
Is The expression of Luiteinizing Hormone Receptor (LHR)
I.3. Objective
testis.
of it.
reduced. Leydig cells from both old control rats and young LH-
old cells produced far less cAMP. cAMP is the classical second
al., 2002).
were divided into 2 kinds, which are low dose (2 Gy and 4 Gy)
While the lower doses (2 and 4 Gy) did not show any
decreasing.
mass, red blood cell production, male hair pattern, libido and
1994).
Advanced glication End-product has a receptor for creating
between genes for class II and III (Sugaya et al., 1994). AGEs
al., 2001).
Figure 3. AGE-RAGE interaction and NF-kB leading to
oxidant stress and generation of pro-inflammatory cytokines
(Singh et al., 2001).
al., 1995).
flesh is sweet and the skin has many benefits and one of it is
resistance.
II.2. Basic Theory
Explanation :
: getting of
: binding
--- : preventing decrease
: caused
: decrease
Shortly, Advanced Glycation End-product could be found in
cell. Another case ROS has effect for lipid membrane will
Gamma Mangosteen
Explanation;
: Become (encounter)
: affects
: applied
laboratory study.
2015.
exclusion criteria.
Mangostin 5M.
Mangosteen.
1. data collection
a Removal of testis
90 days aged Sprague Dawley mices testicle with the
weight around 200 gram will be fasted for about 10 hours
before the taking of testis. The mice is anestesized with
0,3ml/100grBB HCL cetamine i.m. After that, the four leg
of the mice fixed with rope in the surgery table. Fur in
the area abdomen then drabbled with with wet cotton and
shaved until the skin in the surgery area is revealed.
The area then sterilized with alcohol and incisied for
about 2 cm long along the stomach center line with
scalpel. Following that, the specimen then peritoneum
incisied for about 1,5-2 cm long. Using a pair of curving
tweezers and small scissors, a cutlet for about 1 cm long
and 0,5 inch anterior towards the genital area was made
in the center line along the lower abdomen. The mices
skin was so loose that it can be opened to the right and
left direction to take every testis from just one
incision. Both vas deferens will then revealed from one
side of the testis. The left vas deferens then gently
gripped with pliers and lifted partially so that the
cutlet can be observed more clearly.
Vas deferense then ligated and cut in a way that has
explained above. After the testis had lifted, the cut
area then cleaned and observed if there is any
hemorrhage. After that, he peritoneum and skin were sewed
back with thread that can be absorbed by the body. The
scar then applied with iodine until dried. Lastly,
termination was done by applying an extended dose of
anesthesia.
b Leydig Isolation and Cell Culture
1 Prepare testicle and wash with PBS twice. After
that, mince it manually.
2 Prepare 0,25mg/ml Collagenase; 7ml per 1 mice.
Drop a few in the minced testicle and mince it
chemically.
3 The result from previous action then will be put
in 37C incubation tube (2 mices in a 50 conical
tube, and then divided into 2)
4 Prepare 28ml DMEM. The result from the incubation
process then will be homogenized. After 2 minutes,
put supernatant
5 Supernatan will then be 1250rpm centrifuged for 7-
10 minutes, which will be resulted in pellets.
6 Throw away the supernatant and add 2ml DMEM in the
pellets. After that, 1250rpm centrifuge the
specimen for 7-10 minutes twice. Throw away the
Supernatant, take the pellets out, then add 500l
7 Prepare 60%, 40%, 34%, 26%, and 21% gradients (do
not let them become mixed up)
8 Put the pellets, which has been added with 500l
on the top of gradients.
9 Wing centrifuged with 2750rpm for 30-45 minutes.
10 Take the cells between 60% and 40%.
11 Centrifugation will be resulted in pellets. Wash
with PBS, centrifuge it again with the speed of
2000rpm.
12 Put MK DMEM + 1-2cc.
13 Put on TCD gelatin.
c) Post isolation and cell culture
1)Add AGE-BSA 200g/ml to group 2 and AGE-BSA
200g/ml + Gamma-mangostin 5M to group 3.
2)Homogenisation of each groups.
3)Centrifuge it, then take the supernatan.
4)The result from previous action then will be put
in -80C incubation for 2 weeks.
sample.
b There is a test of average characteristic comparison on
One of the requirements one way ANOVA tes has not fulfill
comparison is p<0,05.
with the data normality test, which was Shapiro Wilk test. The
Group 1 = control
Group 2 = AGE-BSA 200g/ml
Group 3 = AGE-BSA 200g/ml + Gamma-mangostin 5M
5
Looking on the table above P value is 0,10 pg/ 10 /24h, it is
differences (p=0,10).
Difference(LSD) analysis.
IV. 2. Discussion
1(7,78 pg/dl) and the highest on group 3(8,06 pg/dl) and group
this resulted in a same way with the theory that stated that
ability as an antioxydant.
the writer argumented two things, which are first, with AGE
cells to radiation.
receptor that used Adult male (90 to 120 days old) Sprague
the contrary.
1.8-kb band of LHR mRNA was detected 5 and 15 days after EDS,
whereas the longer forms (6.8, 4.2, and 2.7 kb) of the message
they lose the ability to convert MTT into purple colored. Thus
V. 1. Conclusion
V. 2.Suggestion
Schmidt AM, Yan SD, Yan SF, Stern, DM 2001, The multiligand
receptor RAGE as a progression factor amplifying immune
and inflammatory responses. J Clin Invest, vol.108,
pp.949955
ATTACHMENT 1.
Descriptives
LH Receptor
AGE-BSA 200g/ml +
Gamma-mangostin 5M 3 8,0680000 ,25646
ATTACHMENT 2.
Tests of Normality
Shapiro-Wilk
AGE-BSA 200g/ml +
,999 3 ,94
Gamma-mangostin 5M
ANOVA
Expression_of_LHR
4,210 2 6 ,072
ATTACHMENT 4.
Multiple Comparisons
Dependent Variable:
Expression_of_LH_
Receptor
LSD
(I) Kelompok (J) Kelompok Mean Difference (I-J) Std. Error Sig. Lower Bound Upper Bound