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Presented by:
Maria Camila Vélez Osorio
Valentina Vela Arbeláez
Universidad Pontificia Bolivariana
Tercer semestre medicina
Apoptosis
Apoptosis is a type of cell death
that multicellular organisms use
to kill damaged or unnecessary
cells in a perfectly controlled way
that minimizes damage to
neighboring cells. The resulting
cellular debris, which are always
surrounded by a plasma
membrane, are eliminated
phagocytosis, avoiding
inflammation in that area.
Ganoderma
Lucidum
Anti-inflamatory
Antioxidant
- Stimulate inmune system
- Improves symptoms of
diabetic patients
- Promotes regeneration of
liver cells
Decreases insulin resistence
and regulates metabolism
Objetivo general
In this study, we investigated the effects
of FYGL o STZ-induced apoptosis of INS-1
cells and its underlying mechanisms.
Ensayo de
viabilidad celular
Métodos
Ensayo cell
counting kit 8
(CCK8)
Células INS-1: 96 pocillos a
una densidad de 1 x 104
células
→ Se incubaron a [FYGL]
- 0,50,100,200,500 y 1000
microgramos/ml por 24h
STZ (0,3mmol/L) y FYGL por
24h
10microlitros
Lector
microplacas
Ensayo de
apoptosis celular
Células INS-1 → Placas de 6 pocillos
[FYGL] 4h
STZ (0,3mM) y [FYGL] 24h adicionales
Tripsina al 0,05%
PBS preenfriado
Suspensión celular con anexina y
PI → 15 min incubados a baja Tº
Citometría de flujo
ELISA
Técnica serológica en la que
se identifica ya sea un
antígeno o anticuerpo al
utilizar un antígeno o
anticuerpo que fue absorbido
a una fase sólida o una
superficie
Enzima que interactúa con
sustrato para colorear la solución
Color Ag o Ac
Este análisis fue usado
en el estudio para
determinar la insulina
recolectada del
sobrenadante celular
Western Blot
Este análisis fue usado en el
estudio para determinar los
niveles de Western blot p38,
p-p38, ERK, p-ERK, JNK,y
p-JNK
Detecta proteínas específicas
con ayuda de electroforesis y
reconocimiento de anticuerpos
Inducción de la
apoptosis
Figura 2a
Resultados
Cell apoptótica:
Condensación de la
cromatina
Fluorocromos:
Visualizan cromatina
Figura 6a
Figura 7a
Figura 8b
FYGL podría mejorar la secreción de insulina aumentando la
expresión de PDX-1 en las células INS-1 dañadas por la STZ
The discusion
Author Approach
Matches
the
study?
14,15
Shen GX. Zheng S, et
al
During diabetes mellitus, the preservation of β
cells is very important. The growing evidences
have suggested that oxidative stress can be
induced by hyperglycemia and some other risk
factors for diabetes
5 Gerber PA, Rutter GA
Oxidative stress is considered to be one of the
critical pathological events that induce β-cell
damage, and impair the ability of insulin
secretion
19
Saini KS, Thompson
C, Winterford CM, et
al
STZ at low concentrations (10–20 mM) can
cause β- cells apoptosis, while at high doses (30
mM), induce β cells necrosis
Conclusions
● FYGL can improve β cells viability and protect β cells
against STZ-induced cell apoptosis and dysfunction.
● FYGL can improve the insulin secretion by decreasing
the activation of JNK, and then increasing the level of
Pdx-1.
● These findings help ones understanding the
mechanisms of FYGL against pancreatic beta cell
damage in diabetes, and suggest that FYGL could be
used as a promising agent for management of diabetes
mellitus.
Valentina Vela A.
María Camila Vélez Osorio

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A proteoglycan extract from Ganoderma Lucidum protects pancreatic beta cells against STZ inducen apoptosis

  • 1. Presented by: Maria Camila Vélez Osorio Valentina Vela Arbeláez Universidad Pontificia Bolivariana Tercer semestre medicina
  • 2. Apoptosis Apoptosis is a type of cell death that multicellular organisms use to kill damaged or unnecessary cells in a perfectly controlled way that minimizes damage to neighboring cells. The resulting cellular debris, which are always surrounded by a plasma membrane, are eliminated phagocytosis, avoiding inflammation in that area.
  • 3. Ganoderma Lucidum Anti-inflamatory Antioxidant - Stimulate inmune system - Improves symptoms of diabetic patients - Promotes regeneration of liver cells Decreases insulin resistence and regulates metabolism
  • 4. Objetivo general In this study, we investigated the effects of FYGL o STZ-induced apoptosis of INS-1 cells and its underlying mechanisms.
  • 5. Ensayo de viabilidad celular Métodos Ensayo cell counting kit 8 (CCK8) Células INS-1: 96 pocillos a una densidad de 1 x 104 células → Se incubaron a [FYGL] - 0,50,100,200,500 y 1000 microgramos/ml por 24h STZ (0,3mmol/L) y FYGL por 24h 10microlitros Lector microplacas
  • 6. Ensayo de apoptosis celular Células INS-1 → Placas de 6 pocillos [FYGL] 4h STZ (0,3mM) y [FYGL] 24h adicionales Tripsina al 0,05% PBS preenfriado Suspensión celular con anexina y PI → 15 min incubados a baja Tº Citometría de flujo
  • 7. ELISA Técnica serológica en la que se identifica ya sea un antígeno o anticuerpo al utilizar un antígeno o anticuerpo que fue absorbido a una fase sólida o una superficie Enzima que interactúa con sustrato para colorear la solución Color Ag o Ac Este análisis fue usado en el estudio para determinar la insulina recolectada del sobrenadante celular
  • 8. Western Blot Este análisis fue usado en el estudio para determinar los niveles de Western blot p38, p-p38, ERK, p-ERK, JNK,y p-JNK Detecta proteínas específicas con ayuda de electroforesis y reconocimiento de anticuerpos
  • 9. Inducción de la apoptosis Figura 2a Resultados Cell apoptótica: Condensación de la cromatina Fluorocromos: Visualizan cromatina
  • 12. Figura 8b FYGL podría mejorar la secreción de insulina aumentando la expresión de PDX-1 en las células INS-1 dañadas por la STZ
  • 13. The discusion Author Approach Matches the study? 14,15 Shen GX. Zheng S, et al During diabetes mellitus, the preservation of β cells is very important. The growing evidences have suggested that oxidative stress can be induced by hyperglycemia and some other risk factors for diabetes 5 Gerber PA, Rutter GA Oxidative stress is considered to be one of the critical pathological events that induce β-cell damage, and impair the ability of insulin secretion 19 Saini KS, Thompson C, Winterford CM, et al STZ at low concentrations (10–20 mM) can cause β- cells apoptosis, while at high doses (30 mM), induce β cells necrosis
  • 14. Conclusions ● FYGL can improve β cells viability and protect β cells against STZ-induced cell apoptosis and dysfunction. ● FYGL can improve the insulin secretion by decreasing the activation of JNK, and then increasing the level of Pdx-1. ● These findings help ones understanding the mechanisms of FYGL against pancreatic beta cell damage in diabetes, and suggest that FYGL could be used as a promising agent for management of diabetes mellitus.