Tag Archives: immunohistochemistry

Orthodontic force: Immunohistochemistry versus hematoxylin-eosin

J. Pharm. Pharmacogn. Res., vol. 10, no. 4, pp. 695-700, July-August 2022.

Short Communication

Histological analyses of orthodontic force in Cavia porcellus: Comparison between immunohistochemistry and hematoxylin-eosin

[Análisis histológicos de la fuerza ortodóncica en Cavia porcellus: Comparación entre inmunohistoquímica y hematoxilina-eosina]

Erliera Sufarnap1,3*, Syafruddin Ilyas2, Nazruddin Nazruddin3, Deddi P. Putra4, Aditya Rachmawati3

1Doctorate Program, Faculty of Dentistry, Universitas Sumatera Utara, Medan, Indonesia.

2Department of Biology, Faculty of Mathematics and Natural Science, Universitas Sumatera Utara, Medan, Indonesia.

3Department of Orthodontic, Faculty of Dentistry, Universitas Sumatera Utara, Medan, Indonesia.

4Faculty of Pharmacy, Universitas Andalas, Padang, Indonesia.

*E-mail: erliera@usu.ac.id

Abstract

Context: Histological quantification of osteoclasts and osteoblasts can evaluate biological responses to orthodontic tooth movement. Histological analysis of bone samples can be technically challenging.

Aims: To evaluate the differences between hematoxylin and eosin (HE) staining and immunohistochemistry (IHC) in quantifying osteoblast and osteoclast cells following the application of static orthodontic force.

Methods: Orthodontic force was applied using a rubber separator around the maxilla incisor of Cavia porcellus. Tooth samples were taken at 0, 4, 8, 14, 21, and 28 days after applying orthodontic force. HE and IHC staining quantify osteoblast and osteoclast cells in the alveolar bone. IHC staining, i.e., Tartrate-resistant acid phosphatase (TRAP) staining, was used to identify osteoclasts, and osteocalcin (OCN) staining was used to identify osteoblasts.

Results: Significantly higher numbers of osteoclasts and osteoblasts were observed with IHC compared to HE staining (p<0.05). Significant positive linear correlations in the numbers of osteoclasts (r = 0.757) and osteoblasts (r = 0.622) identified were observed between IHC and HE staining.

Conclusions: The results of this study indicate HE staining may represent an acceptable alternative method of quantifying osteoclasts and osteoblasts in the preliminary research of orthodontic tooth movement (OTM).

Keywords: hematoxylin; immunohistochemistry; orthodontic; osteocalcin; tartrate-resistant acid phosphatase.

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Resumen

Contexto: La cuantificación histológica de osteoclastos y osteoblastos puede evaluar las respuestas biológicas al movimiento dental ortodóncico. El análisis histológico de muestras de hueso puede ser técnicamente desafiante.

Objetivos: Evaluar las diferencias entre la tinción con hematoxilina y eosina (HE) y la inmunohistoquímica (IHC) en la cuantificación de células de osteoblastos y osteoclastos después de la aplicación de fuerza ortodóncica estática.

Métodos: Se aplicó fuerza de ortodoncia utilizando un separador de goma alrededor del incisivo maxilar de Cavia porcellus. Se tomaron muestras de dientes a los 0, 4, 8, 14, 21 y 28 días después de aplicar la fuerza de ortodoncia. La tinción con HE e IHC cuantifica las células de osteoblastos y osteoclastos en el hueso alveolar. Se usó tinción IHC, es decir, tinción con fosfatasa ácida resistente a tartrato (TRAP), para identificar osteoclastos, y tinción con osteocalcina (OCN) para identificar osteoblastos.

Resultados: Se observaron números significativamente más altos de osteoclastos y osteoblastos con IHC en comparación con la tinción con HE (valor de p<0,05). Se observaron correlaciones lineales positivas significativas en el número de osteoclastos (r = 0,757) y osteoblastos (r = 0,622) identificados entre la tinción IHC y HE.

Conclusiones: Los resultados de este estudio indican que la tinción HE puede representar un método alternativo aceptable para cuantificar osteoclastos y osteoblastos en la investigación preliminar del movimiento dental ortodóncico (OTM).

Palabras Clave: fosfatasa ácida tartrato resistente; hematoxilina; inmunohistoquímica; ortodoncia; osteocalcina.

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Citation Format: Sufarnap E, Ilyas S, Nazruddin N, Putra DP, Rachmawati A (2022) Histological analyses of orthodontic force in Cavia porcellus: Comparison between immunohistochemistry and hematoxylin-eosin. J Pharm Pharmacogn Res 10(4): 695–700.
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© 2022 Journal of Pharmacy & Pharmacognosy Research (JPPRes)

Sisyrinchium palmifolium effects on colitis-associated colon cancer

J. Pharm. Pharmacogn. Res., vol. 10, no. 4, pp. 595-604, July-August 2022.

Original Article

Effect of 1,4-naphthoquinone from Sisyrinchium palmifolium L. extract on in vivo Ki-67 expression and in silico CDK1, CDK2, CDK4 on colitis-associated colon cancer

[Efecto de la 1,4-naftoquinona del extracto de Sisyrinchium palmifolium L. sobre la expresión de Ki-67 in vivo y CDK1, CDK2, CDK4 in silico en el cáncer de colon asociado a colitis]

Roihatul Muti’ah1, Agustina T. Endharti2,3, Muhammad F. Wafi4*

1Departement of Pharmacy, Faculty of Medical and Health Sciences, Maulana Malik Ibrahim State Islamic University of Malang, Indonesia.

2Department of Parasitology, Faculty of Medicine, Brawijaya University, Malang, East Java, Indonesia.

3Biomedical Central Laboratory, Faculty of Medicine, Brawijaya University, Indonesia.

4Master Program in Biomedical Science, Faculty of Medicine, Brawijaya University, Malang, East Java, Indonesia.

*E-mail: farid_wafi@student.ub.ac.id, dr.faridwafi@gmail.com

Abstract

Context: Medicinal plants can be used as an option for the prevention and reduction of cancer cell resistance and its side effects. Sisyrinchium palmifolium L. is thought to have anti-cancer activity with a compound content of 1,4-naphthoquinone.

Aims: To determine the effect of S. palmifolium extract (SPE) with the main compound 1,4-naphthoquinone on Ki-67 expression by in vivo, and CDK1, CDK2, and CDK4 activity by in silico in colonic epithelial cells of BALB/ c mice induced by azoxymethane (AOM) dextran sodium sulfate (DSS).

Methods: Dayak onion (S. palmifolium) was extracted using 96% ethanol as a solvent. The S. palmifolium extract was then made into tablet form by the wet granulation method. Mice that had been induced with AOM-DSS were given S. palmifolium extract therapy. Twenty samples were used, which were divided into five groups. Mice colon tissue was assessed using Ki-67 immunohistochemistry. This study also used the in silico method to see the effect of 1,4-naphthoquinone compounds from S. palmifolium extract on the expression of CDK1, CDK2 and CDK4 with PDB codes 6GU6, 6GUC, and 1GIH.

Results: Ki-67 expression values were 26 ± 6.51 cells at low dosages, 15 ± 1.73 cells at moderate doses, and 11 ± 1.04 cells at high doses. Between the test groups, there was a statistical differences (p<0.05) with the Post Hoc Mann-Whitney test. At the 6GUC receptor, the mean rerank score of the 1,4-naphtoquinone molecule, which was closest to the native ligand, was -54.6572 ± 2.2722 and -90.5455 ± 1.6524kcal/mole. The steric bond on the amino acid lys 33 (A), which exclusively occurs at the 6GUC receptor, was the only commonality of contact.

Conclusions: 1,4-Naphthoquinone from Sisyrinchium palmifolium L. extract could decrease Ki-67 expression by in vivo, which cloud induce a decrease in epithelial cells proliferation in colon cancer, but has no potential as an inhibitor activity of CDK1, CDK2, and CDK4 by in silico.

Keywords: Dayak onion; immunohistochemistry; 1,4-naphthoquinone.

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Resumen

Contexto: Las plantas medicinales pueden usarse como una opción para la prevención y reducción de la resistencia de las células cancerosas y sus efectos secundarios. Se cree que Sisyrinchium palmifolium L. tiene actividad anticancerígena con un contenido compuesto de 1,4-naftoquinona.

Objetivos: Determinar el efecto del extracto de S. palmifolium (SPE) con el compuesto principal 1,4-naftoquinona sobre la expresión de Ki-67 in vivo y la actividad de CDK1, CDK2 y CDK4 in silico en células epiteliales colónicas de ratones BALB/c inducida por azoximetano (AOM) dextrano sulfato de sodio (DSS).

Métodos: La cebolla de Dayak (S. palmifolium) se extrajo usando etanol al 96% como solvente. Luego, el extracto de S. palmifolium se transformó en forma de tableta mediante el método de granulación en húmedo. Los ratones que habían sido inducidos con AOM DSS recibieron terapia con extracto de S. palmifolium. Se utilizaron veinte muestras, las cuales se dividieron en cinco grupos. El tejido de colon de ratones se evaluó usando inmunohistoquímica Ki-67. Este estudio también usó el método in silico para ver el efecto de los compuestos de 1,4-naftoquinona del extracto de S. palmifolium sobre la expresión de CDK1, CDK2 y CDK4 con los códigos PDB 6GU6, 6GUC y 1GIH.

Resultados: Los valores de expresión de Ki-67 fueron 26 ± 6,51 células a dosis bajas, 15 ± 1,73 células a dosis moderadas y 11 ± 1,04 células a dosis altas. Entre los grupos de prueba, hubo diferencias estadísticas (p<0,05) con la prueba Post Hoc Mann-Whitney. En el receptor 6GUC, la puntuación de reclasificación media de la molécula de 1,4-naftoquinona, que era la más cercana al ligando nativo, fue -54,6572 ± 2,2722 y -90,5455 ± 1,6524 kcal/mol. El enlace estérico en el aminoácido lys 33 (A), que ocurre exclusivamente en el receptor 6GUC, fue el único elemento común del contacto.

Conclusiones: La 1,4-naftoquinona del extracto de Sisyrinchium palmifolium L. podría disminuir la expresión de Ki-67 in vivo, lo que induce una disminución en la proliferación de células epiteliales en el cáncer de colon, pero no tiene potencial como inhibidor de la actividad de CDK1, CDK2 y CDK4 in silico.

Palabras Clave: cebolla Dayak; inmunohistoquímica; 1,4-naftoquinona.

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Citation Format: Muti’ah R, Endharti AT, Wafi MF (2022) Inhibition of Sisyrinchium palmifolium L. ethanolic extract on CDK1, CDK2, CDK4 and Ki-67 expression on colitis-associated colon cancer. J Pharm Pharmacogn Res 10(4): 595–604.
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© 2022 Journal of Pharmacy & Pharmacognosy Research (JPPRes)

Recovery of gastrocnemius muscle cells by Vitis gracilis



J Pharm Pharmacogn Res 10(2): 303-309, 2022.

Original Article

Effect of Vitis gracilis Wall (gagatan harimau) in the recovery of gastrocnemius muscle cells and cytochrome c expression of Mus musculus

[Efecto de Vitis gracilis Wall (gagatan harimau) en la recuperación de las células del músculo gastrocnemio y la expresión del citocromo c de Mus musculus]

Nila Zusmita Wasnis1, Syafruddin Ilyas1*, Salomo Hutahaean1, Ramlan Silaban2, Putri C. Situmorang1

1Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara, Medan, Indonesia.

2Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Negeri Medan, Medan, Indonesia.

*E-mail: syafruddin6@usu.ac.id

Abstract

Context: Physical exercise with maximum intensity to fatigue can cause impaired immunity. Gastrocnemius muscle damage is found in eccentric activities in sports. Vitis gracilis has been proven as a healing agent for degenerative diseases.

Aims: To determine whether the V. gracilis Wall extract can recovery of gastrocnemius muscle cells of Mus musculus through reduced inflammatory cells and cytochrome c expression.

Methods: The studies turned into performed in 5 groups of remedies withinside the study. The remedy groups consisted of T1: Negative control (no treatment), T2: Positive control (Swim Until the First Sink/SUFS) + Vit. C 0.2 mg/kg BW, T3: SUFS + 50 g/kg BW of V. gracilis, T4: SUFS + 75 g/kg BW of V. gracilis and T5: SUFS + 100 g/kg BW of V. gracilis. Mice were dissected and then the leg were taken for analysis of gastrocnemius muscle cells in immunohistochemistry.

Results: There was a significant difference (p<0.01) using the ANOVA one-way test on inflammatory cells in gastronomeus muscle tissue of Mus musculus and cytochrome c expression. V. gracilis 100 mg/kg BW can repair and reduce inflammatory cells in the gastronomic muscle tissue. The expression of cytochrome c protein becomes weaker when it is increased.

Conclusions: V. gracilis can recover gastrocnemius muscle cells of Mus musculus through reduced inflammatory cells and cytochrome c expression because of swim exercise.

Keywords: cytochrome c; gastrocnemius; muscle cells; immunohistochemistry; plant extracts.

Resumen

Contexto: El ejercicio físico con máxima intensidad hasta la fatiga puede provocar una alteración de la inmunidad. El daño del músculo gastrocnemio se encuentra en actividades excéntricas en los deportes. Se ha demostrado que la Vitis gracilis es un agente curativo de enfermedades degenerativas.

Objetivos: Determinar si el extracto de pared de V. gracilis puede recuperar las células del músculo gastrocnemio de Mus musculus mediante la reducción de las células inflamatorias y la expresión del citocromo c.

Métodos: Los estudios se realizaron en 5 grupos de remedios dentro del estudio. Los grupos de remedios consistieron en T1: Control negativo (sin tratamiento), T2: Control positivo (Nadar hasta el primer lavabo/SUFS) + Vit. C 0,2 mg/kg BW, T3: SUFS + 50 g/kg BW de V. gracilis, T4: SUFS + 75 g/kg BW de V. gracilis y T5: SUFS + 100 g/kg BW de V. gracilis. Se diseccionaron los ratones y luego se tomaron las patas para analizar las células del músculo gastrocnemio en inmunohistoquímica.

Resultados: Hubo una diferencia significativa (p<0.01) usando la prueba ANOVA unidireccional en células inflamatorias en tejido muscular gastronomeo de Mus musculus y expresión del citocromo c. V. gracilis 100 mg/kg de peso corporal puede reparar y reducir las células inflamatorias del tejido muscular gastronómico. La expresión de la proteína del citocromo c se debilita cuando esta aumenta.

Conclusiones: V. gracilis puede recuperar las células del músculo gastrocnemio de Mus musculus mediante la reducción de las células inflamatorias y la expresión del citocromo c debido al ejercicio de natación.

Palabras Clave: células musculares; citocromo c; extractos de plantas; gastrocnemio; inmunohistoquímica.

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Citation Format: Wasnis NZ, Ilyas S, Hutahaean R, Silaban R, Situmorang PC (2022) Effect of Vitis gracilis Wall (gagatan harimau) in the recovery of gastrocnemius muscle cells and cytochrome c expression of Mus musculus. J Pharm Pharmacogn Res 10(2): 303–309.

© 2022 Journal of Pharmacy & Pharmacognosy Research (JPPRes)

Effect of sikkam leaves on hyperglycemic sperm



J Pharm Pharmacogn Res 10(2): 270-278, 2022.

Original Article

Effect of sikkam (Bischofia javanica Blume) ethanolic extract on the quality and quantity of hyperglycemic rat sperm

[Efecto del extracto etanólico de sikkam (Bischofia javanica Blume) sobre la calidad y cantidad de esperma de rata hiperglucémico]

Syafruddin Ilyas*, Salomo Hutahaean, Rahmat S.H. Sinaga, Putri C. Situmorang

Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara, Medan, Indonesia.

*E-mail: syafruddin6@usu.ac.id

Abstract

Context: Hyperglycemia causes diabetes mellitus (DM), abnormal metabolism, oxidative stress, and chronic complications such as impotence. Hyperglycemia causes testicular atrophy and stromal cell, seminiferous tubular damage, and spermatogenic cells. Bischofia javanica Blume is a plant that is used for the treatment of various chronic conditions and has traditionally by the people of Indonesia as a diabetes medicine.

Aims: To determine the effect of B. javanica extract on the increase in the quality and quantity of sperm of hyperglycemic rats.

Methods: The treatment groups consisted of; G0: negative control (-), G1: positive control (DM: alloxan induction + standard feed), G2-G4: DM + 300, 600 and 900 mg/kg BW of B. javanica leaves ethanol extract, respectively, and G5: DM + glibenclamide 0.5 mg/kg BW. Rats were dissected, and then the testes were taken to analyze sperm quantity and quality and immunohistochemistry.

Results: There was a significant difference (p<0.05) in testes volume, sperm concentration and sperm motility in hyperglycemic rats. The decrease in caspase 3 expression and apoptosis was accompanied by an increase in the dose of the highest B. javanica ethanol extract, and it was seen that testicular histology in groups G4 (900 mg/kg) and G5 (glibenclamide) could improve testicular histology like in the control group (G0).

Conclusions: B. javanica can improve the quality and quantity of hyperglycemic rats’ sperm and also reduce apoptosis via caspase 3 in the histology of testis.

Keywords: apoptosis; caspase 3; hyperglycemia; immunohistochemistry; plant extracts; sperm.

Resumen

Contexto: La hiperglucemia causa diabetes mellitus (DM), metabolismo anormal, estrés oxidativo y complicaciones crónicas como impotencia. La hiperglucemia causa atrofia testicular y células estromales, daño tubular seminífero y células espermatogénicas. Bischofia javanica Blume es una planta que se utiliza para el tratamiento de diversas afecciones crónicas y tradicionalmente la gente de Indonesia la ha utilizado como medicamento para la diabetes.

Objetivos: Determinar el efecto del extracto de B. javanica sobre el aumento de la calidad y cantidad de espermatozoides de ratas hiperglucémicas.

Métodos: Los grupos de tratamiento consistieron en; G0: control negativo (-), G1: control positivo (DM: inducción de aloxano + alimentación estándar), G2-G4: DM + 300, 600 y 900 mg/kg de peso corporal de extracto etanólico de hojas de B. javanica, respectivamente, y G5: DM + glibenclamida 0,5 mg/kg de peso corporal. Se diseccionaron ratas y luego se tomaron los testículos para analizar la cantidad y calidad de los espermatozoides y la inmunohistoquímica.

Resultados: Hubo una diferencia significativa (p<0,05) en el volumen de los testículos, la concentración de espermatozoides y la motilidad de los espermatozoides en ratas hiperglucémicas. La disminución de la expresión de caspasa 3 y apoptosis fue acompañada por un aumento en la mayor dosis del extracto, y se observó mediante histología una mejoría testicular en los grupos G4 (900 mg/kg) y G5 (glibenclamida) similar al grupo control (G0).

Conclusiones: B. javanica puede mejorar la calidad y cantidad de esperma de ratas hiperglucémicas y también reducir la apoptosis vía caspasa 3 en la histología de testículos.

Palabras Clave: apoptosis; caspasa 3; esperma; extractos de plantas; hiperglucemia; inmunohistoquímica.

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Citation Format: Ilyas S, Hutahaean S, Sinaga RSH, Situmorang PC (2022) Effect of sikkam (Bischofia javanica Blume) ethanolic extract on the quality and quantity of hyperglycemic rat sperm. J Pharm Pharmacogn Res 10(2): 270–278.

© 2022 Journal of Pharmacy & Pharmacognosy Research (JPPRes)

Insulin expression and insulitis degree of diabetic rats


J Pharm Pharmacogn Res 9(5): 598-608, 2021.

Original article

Insulin expression and insulitis degree of diabetic rats after giving sikkam leaves (Bischofia javanica Blume)

[Expresión de insulina y grado de insulitis de ratas diabéticas después de administrar hojas de sikkam (Bischofia javanica Blume)]

Cheryl GP. Rumahorbo, Salomo Hutahaean*, Syafruddin Ilyas

Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara, Medan, Indonesia.

*E-mail: salomo@usu.ac.id

Abstract

Context: Gallic acid and quercetin in sikkam leaves (Bischofia javanica) have scientific potential as antidiabetic agents. Quercetin suppresses hyperglycemia by inhibiting active glucose transport. Meanwhile, gallic acid acts as an antidiabetic is closely related to its antioxidant properties. Antioxidant compounds neutralize cells that experience oxidative stress by donating their hydrogen atoms.

Aims: To analyze the degree of insulitis and insulin expression in the pancreatic cells of rats induced diabetes mellitus (DM) after giving B. javanica leaves extract.

Methods: The treatment groups consisted of G0: negative control (-); G1: positive control (alloxan induction + standard feed); G2: alloxan induction + 300 mg/kg BW of B. javanica leaves ethanol extract; G3: alloxan induction + 600 mg/kg BW of B. javanica leaves ethanol extract; G4: alloxan induction + ethanol extract of B. javanica leaves 900 mg/kg BW and G5: alloxan induction + glibenclamide 4.5 mg/kg BW. At day 28, the rats were sacrificed, and the pancreatic tissue dissected. This was analyzed for degree of insulitis and insulin expression by anti-insulin antibodies using immunohistochemistry and hematoxylin-eosin.

Results: There was a significant difference (p = 0.000) in insulin expression and insulitis degree. By the increase of B. javanica leaves dose, the insulin expression value also increased, and the degree of insulitis in Langerhans’ islets of DM rats was decreased. Islets of Langerhans in insulin production returned to normal after being given B. javanica ethanol extract 900 mg/kg BW like glibenclamide.

Conclusions: Bischofia javanica ethanol extract increased insulin production and reduced the degree of insulitis in the islets of Langerhans histology. Further research is needed to determine the extent to which this extract protects from the sequelae of diabetes.

Keywords: hypoglycemic agent; hyperglycemia; immunohistochemistry; insulin expression; insulitis; plant extract.

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Resumen

Contexto: El ácido gálico y la quercetina en las hojas de sikkam (Bischofia javanica) tienen potencial científico como agentes antidiabéticos. La quercetina suprime la hiperglucemia al inhibir el transporte activo de glucosa. Mientras tanto, el ácido gálico actúa como antidiabético y está muy relacionado con sus propiedades antioxidantes. Los compuestos antioxidantes neutralizan las células que experimentan estrés oxidativo al donar sus átomos de hidrógeno.

Objetivos: Analizar el grado de insulitis y expresión de insulina en las células pancreáticas de ratas inducidas por diabetes mellitus (DM) tras la administración de extracto de hojas de B. javanica.

Métodos: Los grupos de tratamiento estuvieron constituidos por G0: control negativo (-); G1: control positivo (inducción de aloxano + alimentación estándar); G2: inducción de aloxano + 300 mg/kg de peso corporal de extracto de etanol de hojas de B. javanica; G3: inducción de aloxano + 600 mg/kg de peso corporal de extracto de etanol de hojas de B. javanica; G4: inducción de aloxano + extracto etanólico de B. javanica hojas 900 mg/kg de peso corporal y G5: inducción de aloxano + glibenclamida 4,5 mg/kg de peso corporal. En el día 28, las ratas fueron sacrificadas y el tejido pancreático disecado. Este se analizó para determinar el grado de insulitis y expresión de insulina mediante anticuerpos antiinsulina usando inmunohistoquímica y hematoxilina-eosina.

Resultados: Hubo una diferencia significativa (p = 0,000) en la expresión de insulina y el grado de insulitis. Al aumentar la dosis de hojas de B. javanica, aumentó el valor de expresión de insulina y disminuyó el grado de insulitis en el islotes de Langerhans de ratas con DM. Los islotes de Langerhans en la producción de insulina volvieron a la normalidad después de recibir el extracto etanólico de B. javanica 900 mg/kg de peso corporal al igual que glibenclamida.

Conclusiones: El extracto de etanol de Bischofia javanica aumentó la producción de insulina y redujo el grado de insulitis en la histología de los islotes de Langerhans. Se necesitan más investigaciones para determinar hasta qué punto este extracto protege de las secuelas de la diabetes.

Palabras Clave: agente hipoglucemiante; expresión de insulina; extracto de planta; hiperglucemia; inmunohistoquímica; insulitis.

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Citation Format: Hutahaean S, Ilyas S, Rumahorbo CGP (2021) Insulin expression and insulitis degree of diabetic rats after giving sikkam leaves (Bischofia javanica Blume). J Pharm Pharmacogn Res 9(5): 598–608.

© 2021 Journal of Pharmacy & Pharmacognosy Research (JPPRes)

Sikkim and apoptosis via cytochrome c in aortic



J Pharm Pharmacogn Res 9(3): 313-323, 2021.

Original Article

Apoptosis via cytochrome c in aortic tissue of diabetes mellitus after giving sikkam leaves (Bischofia javanica Blume)

[Apoptosis vía citocromo c en tejido aórtico de diabetes mellitus después de dar hojas de sikkim (Bischofia javanica Blume)]

Syafruddin Ilyas*, Salomo Hutahaean, Rahmat SH. Sinaga, Putri C. Situmorang

Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara, Medan, Indonesia.

*E-mail: syafruddin6@usu.ac.id

Abstract

Context: Diabetes mellitus (DM) is a metabolic hyperglycemia disease caused by abnormalities in insulin secretion and the highest increase in diabetes sufferers that occurred in Southeast Asian countries, including Indonesia. The sikkam leaf (Bischofia javanica) ethanolic extract contains many secondary metabolites, and some of them are quercetin and gallic acid, which have the potential to be used as antidiabetic.

Aims: To analyze apoptosis via cytochrome c in aortic tissue of DM after giving B. javanica leaves.

Methods: The treatment group consisted of G0: Negative control, G1: Positive control (alloxan induction + standard feed), G2: Alloxan induction + 300 mg/kg BW of B. javanica leaves ethanol extract, G3: Alloxan induction + 600 mg/kg BW of B. javanica leaves ethanol extract, G4: Induction of alloxan + ethanol extract of B. javanica leaves 900 mg/kg BW and G5: Induction of alloxan + glibenclamide 0.5 mg/kg BW. Fourteen days later, the rats were dissected, and the aortic tissue was analyzed for apoptosis by tunnel assay and cytochrome c by immunohistochemistry.

Results: There was a statistically significant difference (p<0.01) in the apoptotic value and cytochrome c. Increasing the dose of B. javanica leaves, the cytochrome c expression, and apoptotic value decreases in the aorta of DM rats. The cell shape returned to normal after being given B. javanica ethanol extract 900 mg/kg BW than glibenclamide.

Conclusions: Bischofia javanica ethanolic extract reduced apoptosis via cytochrome c in aortic histology, and this plant can be developed into a diabetic drug candidate.

Keywords: apoptosis; cytochrome c; hyperglycemia; hypoglycemic agents; immunohistochemistry; plant extracts.

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Resumen

Contexto: La diabetes mellitus (DM) es una enfermedad de hiperglucemia metabólica causada por anomalías en la secreción de insulina y el mayor aumento de pacientes con diabetes que se produjo en los países del sudeste asiático, incluida Indonesia. El extracto etanólico de la hoja de Bischofia javanica contiene muchos metabolitos secundarios, y algunos de ellos son la quercetina y el ácido gálico, que tienen el potencial de usarse como antidiabéticos.

Objetivos: Analizar la apoptosis por citocromo c en tejido aórtico de DM tras la administración de hojas de B. javanica.

Métodos: El grupo de tratamiento consistió en G0: Control negativo, G1: Control positivo (inducción de aloxano + alimentación estándar), G2: inducción de aloxano + 300 mg/kg de peso corporal de extracto de etanol de hojas de B. javanica, G3: inducción de aloxano + 600 mg/kg de peso corporal de extracto etanólico de hojas de B. javanica, G4: Inducción de aloxano + extracto etanólico de hojas de B. javanica 900 mg/kg de peso corporal y G5: Inducción de aloxano + glibenclamida 0,5 mg/kg de peso corporal. Catorce días más tarde, las ratas fueron disecadas y el tejido aórtico fue analizado para apoptosis por ensayo de túnel y citocromo c por inmunohistoquímica.

Resultados: Hubo una diferencia estadísticamente significativa (p<0.01) en el valor apoptótico y el citocromo c. Al aumentar la dosis de hojas de B. javanica, la expresión del citocromo c y el valor apoptótico disminuyen en la aorta de ratas DM. La forma de la célula volvió a la normalidad después de recibir extracto de etanol de B. javanica 900 mg/kg de peso corporal que glibenclamida.

Conclusiones: El extracto etanólico de Bischofia javanica redujo la apoptosis a través del citocromo c en histología aórtica y esta planta puede convertirse en un candidato a fármaco diabético.

Palabras Clave: agentes hipoglucemiantes; apoptosis; citocromo c; extractos de plantas; hiperglucemia; inmunohistoquímica.

This image has an empty alt attribute; its file name is jppres_pdf_free.png
Citation Format: Ilyas S, Hutahaean S, Sinaga RSH, Situmorang PC (2021) Apoptosis via cytochrome c in aortic tissue of diabetes mellitus after giving sikkam leaves (Bischofia javanica Blume). J Pharm Pharmacogn Res 9(3): 313–323.

© 2021 Journal of Pharmacy & Pharmacognosy Research (JPPRes)