THE INCREASING OF VEGF EXPRESSION AND RE-EPITHELIALIZATION ON DERMAL WOUND HEALING PROCESS AFTER TREATMENT OF BANANA PEEL EXTRACT (MUSA ACUMINATA COLLA)

Objective: The objective of the study was to determine the efficacy of topical administration of an alcoholic bark extract of Musa acuminata Colla on cutaneous wound healing including expression of VEGF and re-epithelization on dermal in rats. Methods: Model was performed to evaluate the excision model of wound healing activity. Full-thickness excision wounds were made on the back of rat and Musa acuminata Colla extract was topically administered (doses 25%, 50% & 75%). The formation of granulation tissue was observed on day 4, 8, 12 and 16 (post-wound) to indicate VEGF and collagen. The extract increased cellular re-epithelialisation and collagen synthesis at the wound site, shown by increase in VEGF, and total collagen content of granulation tissues. The rate of contraction in wounds was determined by tracing the wound surface onto a transparent graph paper and measuring the surface area by planimetry. Results: From the observation in EBP, all doses were found to have wound healing activity in wound contraction and period of epithelialization. The increasing of VEGF expression was found greater in the extract treated group than control group. The tensile strength of extract treated group was increased significantly. Conclusion: The results indicated that Musa acuminata Colla peel extracts has significant wound healing activity

Pivotal role reelin signaling pathway in the development of tolerance to morphine-induced antinociception

The huge endogenous macromolecule protein responsible for controlling migration and dendritic growth of developing neurons, reelin, has recently been proposed that its signaling pathway modulates synaptic plasticity in the adult rodent brain. This study was carried out to investigate the pivotal role of the reelin signaling pathway in the development of tolerance to morphine induced antinociception. There was evidence that repeated intracerebroventricular administration of reelin’s monoclonal antibody, the competitive inhibitor to reelin – apolipoprotein receptor E2 recombinant, and disabled1 (Dab1) protein inhibitor – MG132, resulted in the inhibition to the development of antinociception tolerance to morphine administration. Furthermore, chronic in vivo administration with morphine caused significance increase of the immunoreactivity (IR) for phosphorylated-Dab1 in the thalamus. These data suggested that persistent activation of reelin signaling pathway due to chronic administration of morphine may be responsible for the development of tolerance to morphine-induced antinociception.Key words: Morphine tolerance, Neuronal plasticity, Opioid receptor, Reelin signalling pathwa

In Vitro Gentamicin Release from Bioactive BHAG(ELENA) Implant Against Staphylococcus aureus

Osteomyelitis is a bone-related infectious desease which is difficult to treat, because the antibiotic reaches the target is lower than the MIC and bacteria can’t be eradicated. This condition can cause the bacteria become resistant. To solve this problem, we can use local antibiotics as BHAG(ELENA) pellets implant, which can release gentamicin (GEN) continuously for more than a day with a concentration greater than MIC. BHAG(ELENA) pellet that have made contain BHA : GEL = 20 : 2 (dry state); GEN 10% and cross-linking with glutaraldehyde (GA) 0,5%, cylindrical weigh 100 mg; 4 mm in diameter and 3.2 mm thick. The release of GEN from BHAG(ELENA) pellet were tested in vitro, by soaking the pellet in phosphate buffer saline pH 7.4 at temperature 370C. The sample were sampled every day until 28 days. Then, the sample were tested by agar diffusion method that contain Staphylococcus aureus. Results showed that inhibition zone diameter greater than MIC GEN to S. aureus. Within 28 days, the release of GEN provide a total activity 99,24%, it showed that after 28 days, the pellets are still actively inhibit the bacterial growth. Furthermore, required to be tested in animal study (in vivo) with a defect in the femoral bone then filled with BHAG(ELENA) pellets as drug delivery system of GEN and bone fillers. Keywords: agar diffusion method, Bovine Hydroxyapatite (BHA), gelatin (GEL), gentamicin (GEN), glutaraldehyde (GA) and drug delivery system (DDS)

The Effectiveness of GABA Agonist in Decreasing Expression of NR2B Subunit of N-Methyl-D-Aspartate (NMDA) Receptor in Neuropathic Mice by Partial Sciatic Nerve Ligation (PSNL) Method

Neuropathy pain is a pain that caused by nerves injury. Nowadays, treatment for neuropathic pain change to drugs that works as GABA agonist and cause reimbalance between excitatory and inhibitory neurotransmitter in central nervous system (CNS). The present study was designed to investigate the effectiveness of gabapentin and baclofen in decreasing N-Methyl-D-Aspartate (NMDA) receptor NR2B subunit activity in neuropathic pain. Fourty mice were divided into 8 groups i.e sham, negative control, gabapentin (10, 30, 100 nmol) and baclofen (1, 10, 30 nmol). Neuropathic pain was induced by ligation of sciatic nerve with Partial Sciatic Nerve Ligation (PSNL) method). Treatments were administrated intrathecally once a day for seven consecutive days, at a week after induction. On day 15th, mice were sacrified and the spinal cord were removed quickly. The expression of NMDA receptor NR2B subunit were examined with imunohistochemistry and data were analyzed by one way anova. The result from this research was gabapentin and baclofen administration significantly decrease expression of NMDA receptor NR2B subunit in mice compared to sham group. The higher the dose, the more effective to decrease the number of neuron that express NR2B. The conclusion of this research was gabapentin and baclofen treat neuropatic pain by decreased the number of NMDA receptor NR2B subunit.Nyeri neuropati merupakan nyeri yang disebabkan oleh trauma atau cedera pada saraf. Saat ini, terapi untuk mengatasi nyeri neuropati beralih pada agonis GABA yang menyebabkan kembalinya keseimbangan antara neurotransmitter eksitatori dan inhibitori di sistem saraf pusat (SSP). Penelitian ini bertujuan untuk membuktikan efektivitas dari gabapentin dan baclofen dalam menurunkan ekspresi dari reseptor N-Methyl-D-Aspartate (NMDA) subunit NR2B pada nyeri neuropati. Empat puluh mencit dibagi kedalam 8 kelompok yaitu: sham, kontrol negatif, gabapentin (10, 30, 100 nmol) dan baclofen (1, 10, 30 nmol). Nyeri neuropati diinduksi dengan ligasi pada saraf sciatic dengan metode Partial Sciatic Nerve Ligation (PSNL). Senyawa uji diberikan setiap hari selama tujuh hari, satu minggu setelah induksi. Pada hari ke-15, mencit dikorbankan dan diambil bagian spinal cord. Ekspresi reseptor NMDA subunit NR2B diamati dengan imunohistokimia dan data dianalisis menggunakan anova satu arah. Hasil dari penelitian ini menunjukkan gabapentin dan baclofen menurunkan ekspresi reseptor NMDA subunit NR2B (dose dependent) dibandingkan kelompok sham. Kesimpulan dari penelitian ini bahwa gabapentin dan baclofen dapat mengatasi nyeri neuropati melalui penurunan aktivitas reseptor NMDA subunit NR2B

THE INFLUENCE OF SODIUM ORTHOVANADATE ON P85 AND GSK-3 EXPRESSIONS TO THE BLOOD GLUCOSE REGULATION OF TYPE 2 DIABETIC MICE (MUS MUSCULUS) MODEL

Objective: The present study was designed to investigate the influence of sodium orthovanadate on the P85 (Regulatory Subunit of PI3-Kinase) and GSK-3 (Glycogen Synthase Kinase 3) expressions in skeletal muscle tissue to the decreasing of blood glucose levels of alloxan-induced diabetic mice.Methods: Mice were divided into 5 groups i. e. (1) naive group, (2) diabetic group and (3-5) orthovanadate-treated diabetic groups at the dose of 16, 32 and 64 mg/kg BW respectively. Diabetic mice model was induced by intraperitoneal administration of alloxan monohydrate at the dose of 200 mg/kgBW. Diabetic state was occurred on day 3 after alloxan injection and then started the treatment of sodium orthovanadate for 7 days. The muscular tissue was harvested on day 10 after treatment and was stained using routine histology staining, haematoxylin-eosin for morphological qualitative analysis and immunohistochemical approaches to observe the expressions of P85 and GSK-3 in skeletal muscle.Results: Diabetes condition was shown by the increasing of fasting blood glucose levels from 59.1 ± 11.2 mg/dL to 310.6 ± 107.2 mg/dL on day 3. Administration of sodium orthovanadate at the dose of 16, 32 and 64 mg/kg BW reduced the fasting blood glucose levels after 7 days treatment (p < 0.001) at diabetic mice significantly. The results of histology staining showed that sodium orthovanadate improved a necrosis in skeletal muscle cells alloxan-induced diabetic mice. On immunohistochemical approaches, sodium orthovanadate might decreased the P85 expressions and increased the GSK-3 expressions in skeletal muscle cells alloxan-induced diabetic mice (p < 0.001).Conclusion: This study reveals that the administration of sodium orthovanadate on diabetic mice led to the attenuation of the PTPase activity. The inhibition cause a decreasing the expression of P85 and increasing of GSK-3 that cause the decreasing of blood glucose levels and an improvement of insulin target cells in the muscle cells.Â

Pengaruh Vanadil Sulfat Terhadap Ekspresi Protein GLUT-4 pada Mencit yang Menderita Diabetes Mellitus

Penelitian ini bertujuan untuk mengetahui pengaruh vanadil sulfat terhadap aktivitas protein GLUT-4 pada otot skelet mencit yang menderita diabetes mellitus akibat induksi streptozotocin. Sebanyak 25 ekor mencit dibagi menjadi 5 kelompok, yaitu kelompok kontrol, diabtes dan perlakuan yang diberi vanadil sulfat dengan dosis 5,30 dan 100 mg/kgBB. Keadaan diabetes diinduksi dengan pemberian streptozotocin dosis 100 mg/kgBB pada hari pertama dan 50 mg/kgBB pada hari ke -14. Pada hari ke-21 terjadi peningkatan kadar glukosa dari 151,4 ± 25,1 mg/dL menjadi 237,1 ± 33,0 mg/dL. Pemberian vanadil sulfat selama 7 hari akan menurunkan kadar glukosa darah secara signifikan (p<0,001). Otot skelet diambil pada hari ke-28 dan dipreparasi dengan hematoksilin-eosin untuk diamati histologinya dan secara imunohistokimia untuk pengamatan aktivitas protein GLUT-4. Hasil pengamatan menunjukkan pemberian vanadil sulfat akan memperbaiki keadaan atropi dan menghambat nekrosis pada sel otot skelet. Pengamatan secara imonohistokimia menunjukkan bahwa vanadil sulfat dapat meningkatkan aktivitas GLUT-4

EFFECTIVENESS AND MECHANISM OF ACTION OF VANADYL SULFATE IN INCREASING β CELL PROLIFERATION OF DM MICE DUE TO STREPTOZOTOCIN INDUCTION

Diabetes mellitus (DM) is a metabolic disorder characterized by hyperglycemia as a result of damage to insulin secretion, insulin action, or both. Vanadyl sulfate is one of the form of vanadium which has begun to be used to treat diabetes in humans. In this study, investigation on the effectiveness of vanadyl sulfate in increasing pancreatic β cell proliferation of diabetes mice due to streptozotocin induction. There were 30 healthy mice were subjected to this experimental study. Findings from this experiment proved that administration of vanadyl sulfate at various doses can significantly increase the amount of pancreatic β cell proliferation and eventually reduced blood glucose levels in a meaningless manner. However, those mice that administrated of vanadyl sulfate at a dose of 30 mg/kgBW had a higher amount of Langerhans Islet than those in the vanadyl sulfate group at a dose of 5 mg/kgBW and 100 mg/kgBW. Similar observations were obtained for the Ki-67 expression. The highest Ki-67 expression was obtained at a dose of 30 mg/kgBW but it decreased with a dose of 100 mg/kgBW of vanadyl sulfate

Preventive Effects of Alpha-Lipoic Acid on Lipopolysaccharide-Induced Endothelial Dysfunction in Rats

Endothelial dysfunction has been considered as one of the key initial events in the pathogenesis of atherosclerosis and other cardiovascular diseases (CVDs). Several studies imply that chronic inflammation and oxidative stress play a critical role in endothelial dysfunction. The present study was designed to evaluate the preventive effect of alpha-lipoic acid (ALA) on lipopolysaccharide (LPS) induced endothelial dysfunction in rat based on malondialdehyde (MDA) level and vascular cell adhesion molecules-1 (VCAM-1) expression. Thirty Wistar rats were administered ALA for 2 weeks in different doses (30, 60, 120 mg.kg-1 BW) 1 hour before LPS 0.5 mg.kg-1 BW i.p challenge. The LPS was injected on 1st, 4th, and 9th day. MDA plasma level was analyzed with spectrophotometer &lambda; 529 nm, and VCAM-1 expression was determined by immunohistochemistry. Pretreatment with ALA for 14 days could decrease plasma MDA level on LPS-induced endothelial dysfunction in rats. However, only one group of ALA doses, 120 mg.kg-1 BW, showed significant difference with LPS untreated group statistically. It was also found that ALA, in all treatment groups, could attenuate VCAM-1 expression. These findings suggest that ALA had a protective effect against endothelial dysfunction and may potentially prevent vascular inflammatory disease

Strategi Pendekatan Mechanism-Based Therapy Pada Nyeri Neuropati Dengan Senyawa Agonis Reseptor Gaba-B Baclofen

Nyeri merupakan rasa atau pengalaman yang menyakitkan karena adanya kerusakan jaringan atau adanya potensi kerusakan jaringan. Lebih dari 90% penyakit selalu dlscrtai oleh rasa nyeri dimana 40% kasus berkembang pada nyeri kronik atau nyeri neuropati. Nyt.ri neuropati teIjadi karena rusaknya atau tidak berfungsinya sistem saraf baik sentral maupun perifer. Nyeri neuropati, seperti yang teIjadi pada nyeri kanker dan nyeri diabetic neuropati, menjadi problem serius karena akan menurunkan kualitas bidup, penurunan produktivitas dan besarnya biaya pengobatan. Pada masa mendatang, penderita nyeri neuropati akan meningkat sejalan dengan meningkatnya prevalensi populasi lanjut usia dan meningkatnya penyakitdegeneratif. Manajemen terapi nyeri neuropati dengan pemberian nonsteroidal anti-inflammatory drugs (NSAIDs) ataupun parasetamol dan bahkan opioids temyata kurang efektif dalam mengatasi nyeri neuropati. Oleh karena itu, altematifterapi dengan mengaktivasi GABA-ergic sistem sebagai antagonis glutamat. memberikan araban strategi pendelqttan terapi penanganan nyeri neuropati. Baclofen merupakan agonis GABA reseptor

Histologi Dorsal Horn Dari Spinal Cord Mencit Yang Mengalami Nyeri Inflamasi Akibat Induksi Cfa (Completed Freuds Adjuvant) Setelah Pemberian Gabapentin Dan Baclofen

Pain is a multidimentional experience, generally most of chronic disease followed by pain incidence. Chronic pain can be caused by inflammation or neuropathic condition and change the imbalance of N-methyl-D-aspartate receptor 2B (NR2B) subunit. This research was an observation of the influence of gabapentin and baclofen to dorsal horn histology in inflammatory-induced chronic pain. Fourty mice Balb-C strain were divided into 8 groups i.e sham, negative control, gabapentin at three different doses (10, 30, 100 nmol) and baclofen at three different doses (1, 10, 30 nmol). Inflammatory condition was induced by intraplantar injection of CFA (Completed Freuds Adjuvant). Gabapentin and baclofen were given intrathecally once a day for seven consecutive days, at a week after CFA injection. Latency time toward thermal stimulus was measured on days 0, 1, 3, 5, 7, 8, 10, 12 and 14 after CFA injection. Paw thickness at the ipsilateral site was also measured on days 0, 1, 2, 3, 4, 5, 6, 7, 8, 10, 12 and 14 after CFA injection. Histology of the dorsal horn of spinal cord tissue was examined by haematoxylline-eosin staining. The result showed that intrathecal injection gabapentin and baclofen significantly increased latency time of mice toward thermal stimulus compared to negative control. Whereas gabapentin and baclofen administration could decrease inflammatory cell, vasodilatation and increase neuron forming of the dorsal horn histology compare to negative control. The conclusion of this research was gabapentin and baclofen administration had antinociceptic effect by increased latency time toward thermal stimulus and recoved histology of dorsal horn from mice with inflammatory painKeywords : Inflammation, CFA, Gabapentin, Baclofen, Dorsal horn.AbstrakNyeri merupakan pengalaman yang multidimensional. Umumnya kebanyakan penyakit kronik selalu disertai dengan nyeri. Nyeri kronik dapat disebabkan oleh inflamasi maupun neuropati dengan patofisiologi yang berhubungan dengan aktivitas reseptor N-methyl-D-aspartate (NMDA) subunit 2B (NR2B). Sampai saat ini pengobatan nyeri kronik menjadi tantangan. Obat yang bekerja sebagai agonis GABA seperti gabapentin dan baclofen dilaporkan mempunyai peranan penting dalam penghambatan proses nyeri. Penelitian ini dilakukan untuk mengetahui pengaruh pemberian gabapentin dan baclofen terhadap histologi dorsal horn pada keadaan nyeri kronik akibat inflamasi. Hal ini bertujuan untuk menjelaskan bagaimana gabapentin dan baclofen dapat digunakan sebagai terapi pada nyeri kronik. Empat puluh mencit dibagi menjadi delapan kelompok, yaitu sham, kontrol negatif, gabapentin dosis 10, 30 dan 100 nmol/mencit serta baclofen dosis 1, 10 dan 30 nmol/mencit. keadaan inflamasi diinduksi oleh injeksi intraplantar CFA (Completed Freuds Adjuvants). Gabapentin dan baclofen diberikan secara intratekal sehari sekali selama tujuh hari, pada hari ketujuh setelah induksi CFA. Waktu ketahanan terhadap stimulus panas diukur menggunakan hot/cold plate pada hari ke-0, 1, 3, 5, 7, 8, 10, 12 dan 14 setelah induksi. Tebal plantar diukur pada hari ke-0, 1, 2, 3, 4, 5, 6, 7, 8, 10, 12 dan 14 setelah induksi. Respon nyeri diamati secara visual seperti mendekatkan kedua tungkai kaki ke depan, menjilat tungkai kaki ke depan, gerakan meliuk, berusaha melompat keluar hot/cold plate,dan menghentakkan tungkai belakang. Histologi bagian dorsal horn dari spinal cord diamati menggunakan pewarnaan haematoxyllin-eosin. Pemberian gabapentin dan baclofen meningkatkan waktu ketahanan terhadap stimulus panas secara signifikan dibandingkan kontrol. Secara histologi, pemberian gabapentin dan baclofen menurunkan sel inflamatori, menurunkan vasodilatasi dan meningkatkan bentukan neuron pada dorsal horn dari spinal cord dibandingkan dengan kontrol. Kesimpulan dari penelitian ini adalah pemberian gabapentin dan baclofen meningkatkan waktu ketahanan terhadap stimulus panas serta memperbaiki histologi dorsal horn dari spinal cord mencit dengan nyeri inflamasi setelah induksi CFA.Kata kunci : Nyeri inflamasi, CFA, Gabapentin, Baclofen, Dorsal hor