PENINGKATAN L-TYPE CALCIUM CHANNEL, KALSIUM INTRASELULER, PAR-1 dan HSP-27 pada UTERUS KELINCI NEW ZEALAND YANG DILAKUKAN
PENJAHITAN MODIFIKASI SURABAYA (KAJIAN BIOMOLEKULER PADA PENANGANAN PERDARAHAN PASCA PERSALINAN KARENA ATONIA UTERI)
SULISTYONO, AGUS
Promotor: Prof. Dr. Kuntoro, dr, MPH
INTRACELLULAR CALCIUM KKA KK Dis K 53/12 Sul p
Copyright© 2012 by Airlangga University Library Surabaya
RINGKASAN
Setiap tahun terdapat 529.000 kematian maternal dan 99% terjadi di negara berkembang, penyebab utamanya adalah perdarahan pasca persalinan (PPP) yaitu sebanyak 25%. Berdasarkan Survei Demografi dan Kesehatan Indonesia pada tahun 1994 terdapat kematian maternal sebanyak 390 per 100.000 kelahiran hidup, tahun 2001 menurun menjadi 307 dan tahun 2007 sebanyak 248. Target angka kematian maternal Millenium Development Goal tahun 2015 menjadi 100 kematian maternal per 100.000 kelahiran hidup. Terdapat 4 penyebab utama PPP, atonia uteri merupakan penyebab terbanyak yaitu 70%, sisanya karena laserasi jalan lahir, kelainan penempelan plasenta dan kelainan darah. Penanganan PPP yang paling penting adalah menghentikan perdarahan sambil memperbaiki keadaan umum ibu, menentukan etiologinya dan memberikan terapi yang sesuai. Kegagalan menentukan etiologi dan penanganan yang tepat menyebabkan perdarahan tidak berhenti dan menimbulkan penyulit yang sulit diatasi. Mekanisme penghentian perdarahan pada PPP berbeda dengan organ lain yang tergantung vasospasme pembuluh darah dan pembekuan darah. Pada PPP penghentian perdarahan terutama karena kontraksi dan retraksi miometrium sehingga menyempitkan dan membuntu lumen pembuluh darah. Bila kontraksi dan retraksi tidak berfungsi dengan baik, perdarahan tidak akan berhenti walaupun faktor pembeku darah normal, sebaliknya walaupun faal pembeku darah tidak baik PPP dapat berhenti asalkan kontraksi dan retraksi uterus berfungsi normal. Kontraksi miometrium dipengaruhi oleh peningkatan kalsium intraseluler yang terutama berasal dari influks melalui L-type calcium channel. Kalsium intraseluler berikatan dengan reseptornya, yaitu kalmodulin membentuk kompleks kalsium kalmodulin yang dapat mengaktivasi Myosin Light Chain Kinase
rangsangan mekano-transduksi yang dapat menimbulkan depolarisasi membran sel sehingga L-type calcium channel membuka dan juga mengaktifkan protein G. Untuk mengkonfirmasi terjadi kontraksi miometrium setelah dilakukan penjahitan kompresi metode Surabaya, dilakukan penelitian eksperimental laboratorium secara cross sectional, single blinded dengan interval waktu ½, 1, 2 dan 24 jam pada model uterus kelinci bunting aterm yang dilakukan operasi pengeluaran janin, irisan uterus dijahit kembali, selanjutnya pada uterus ipsilateral dilakukan penjahitan kompresi modifikasi Surabaya sedang sisi kontralateralnya sebagai kontrol tidak dilakukan. Variabel kontraktilitas miometrium yang diteliti adalah ekspresi L-type calcium channel, PAR-1 dan HSP-27 yang diperiksa secara imunohistokimia serta densitas kalsium intraseluler berdasar intensitasnya di sitoplasma yang diperiksa dengan cara fluoro 3 double staining
SUMMARY
THE INCREASED EXPRESSION OF L-TYPE CALCIUM CHANNEL, PAR-1 AND HSP-27 AND DENSITY OF INTRACELLULAR CALCIUM ON ANIMAL
MODEL OF NEW ZEALAND RABBIT MYOMETRIUM THAT USING CONSERVATIVE SURGICAL PROCEDURE OF SURABAYA SUTURE
COMPRESSION MODIFICATION
(BIOMOLECULAR STUDY IN CONSERVATIVE SURGICAL MANAGEMENT OF PPH DUE TO UTERINE ATONY)
AGUS SULISTYONO
bleeding in many cases of PPH due to atony. The aims of this study is to confirm increasing L-type calcium channel, intracellular calcium, PAR-1 and HSP-27 in Surabaya compression suturing as a mechanotransducer biomolecular pathways to induce myometrial contraction. The study was done from March to August 2011, true experimental, single blinded with time interval were ½, 1, 2 and 24 hours, using term uterine pregnant New Zealand rabbits. Its consists 11 uterine samples of Surabaya methods (cases) and 11 samples without compression suturing technique as controls. The result of this study is significantly increase of all variables observed in cases compare to control groups at 4 time intervals, ½, 1, 2 and 24 hours. At ½ hour observation, the expression of L-type calcium channel are 2,78±1,39 vs 12,89±5,30 (p=0,001); 1 hour 3,93±0,49 vs 14,87±1,05 (p<0,0001); 2 hour 1,48±0,46 vs 10,01±3,21 (p=0,011) dan 24 hour are 2,95±0,18 vs 10,01±3,21 (p<0,0001) cell/100 cell observational field. The densities of intracellular calcium at ½ hour are 1722,52±769,30 vs 2375,26±264,53(p=0,22); at 1 hour observation are 1721,60±426,93 vs 2467,84±209,13 (p=0,001); at 2 hours are 1722,78±449,92 vs 2233,24±283,73 (p=0,005) and at 24 hours are 1527,52±167,63 vs 1988,61±293,73 (p=0,002) intensities/μm2. Expression of PAR-1 are 5,39±1,18 vs 16,36±1,31 (p<0,0001); 12,34±1,81 vs 23,72±2,80 (p<0,0001); 12,71,1,60 vs 23,42±1,38 (p<0,0001) and 12,55±1,60 vs 22,97±1,35 (p<0,0001) cells/observational field. HSP-27 expression are 3,82±1,49 vs 13,15±2,99 (p<0,0001); 4,93±2,67 vs 22,45±4,06 (p<0,0001); 5,20±1,71 vs 21,09±4,06 (p<0,0001) dan 5,41±1,73 vs 21,09±1,92 (p<0,0001) cells/observational field. Conclusion of this study is uterine compression suture (Surabaya modification suture) increasing all variables, L-type calcium channel, PAR-1, HSP-27 expression and calcium intracellular density compare to control. Mechanotransduction of uterine compression trigger uterine myocyte contractility pathway to induce myometrial contraction.
ABSTRACT
THE INCREASED EXPRESSION OF L-TYPE CALCIUM CHANNEL, PAR-1 AND HSP-27 AND DENSITY OF INTRACELLULAR CALCIUM ON ANIMAL
MODEL OF NEW ZEALAND RABBIT MYOMETRIUM THAT USING CONSERVATIVE SURGICAL PROCEDURE OF SURABAYA SUTURE
COMPRESSION MODIFICATION
(BIOMOLECULAR STUDY IN CONSERVATIVE SURGICAL MANAGEMENT OF PPH DUE TO UTERINE ATONY)
AGUS SULISTYONO
Objective : Uterine atony as the common cause of PPH which causing high maternal mortality can indeed be largely rescued by compression with conservative surgical procedure by performing Surabaya suture compression modification (SSCM).
of L-type calcium channel, PAR-1 and HSP-27 and density of intracellular calcium on animal model of New Zealand rabbit myometrium after procedure performance
Material and methods: True experimental single blinded study was done in Airlangga University Veterinary Faculty and Brawijaya University Physiology Department from March – August 2011. Twenty two samples were obtained over this study which applied on aterm pregnant rabbit uterus as animal model. Animal model consisted of rabbit uterus in which practiced on by SSCM in 11 samples after terminating pregnancy and removing pup and placenta as well, and the remainder of another 11 samples without SSCM as the control. The assessment of L-type calcium channel, PAR-1 and HSP-27 expression and density of intracellular calcium on myometrium sample was done respectively by immunohistochemistry and fluoro-2 double staining method at ½,1,2, and 24 hours intervals
Result: At all 4 time intervals the increased expression of L-type calcium channel was observed significantly in SSCM compared to control (p=0.001; p< 0.0001; p=0.011 and p<0.0001), also for intracellular calcium density (p=0.022; p=0.001;p=0.005 and p=0.002). The study showed PAR-1 and HSP-27 expression increasing sharply that statistically analysis indicated both all of p similarly < 0.0001 against control.
Conclusion: SSCM suggested strongly increasing some component (all variable) that absolutely required to myometrium contraction
