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Jazmin Gómez Olarte
Tercer semestre 2020-1
Facultad de medicina, Universidad Pontificia Bolivariana
INTRODUCTION
Phillygenin is a component extracted from Forsythiae Fructus.
Forsythia Suspensa Forsythiae Fructus
Phillygenin
* Detoxify
* Therapeutic effects
INTRODUCTION
TLR4/MYD88/NF-kB pathway
LPS: Lipopolysaccharide
TLR: Toll-like receptors
PHI could inhibit the proinflammatory
reaction induced by LPS, in addition it
could inhibit protein expression in the
TLR4 / MyD88 / NF-κB signal pathway.
OBJECTIVE
Investigate if Phillygenin
(PHI) has a therapeutic
effect on liver fibrosis
caused by inflammation.
MATERIALES Y MÉTODOS
Para qué
Para cultivo de las células
LX2 (línea de las células
estrelladas hepáticas del
humano).
CULTIVO CELULAR Fundamento
Proceso mediante el que
las células pueden
cultivarse en condiciones
controladas.
Fundamento Para qué
ELISA
Enzyme-Linked
ImmunoSorbent Assay
MATERIALES Y MÉTODOS
Detección de los
factores inflamatorios
IL-6, IL-1β y TNF-α.
*Prueba para determinar
[Antígeno] o
[Anticuerpo] con uno de
ellos en fase sólida y el
otro en solución.
* Conjugación enzima-Ac
que genera un producto
detectable.
MATERIALES Y MÉTODOS
Fundamento
* Retrotranscripción de una hebra
de ARN en ADN.
* Uso de transcriptasa inversa.
RT-qPCR
Reacción en cadena de la polimerasa con
transcriptasa inversa
Para qué
Determinación de la expresión de
colágeno I y α-SMA (secreción de
actina del músculo liso α) y para
verificación del mecanismo de PHI en
la ruta de señal TLR4 / MyD88 / NF-κB
MATERIALES Y MÉTODOS
Usado para la
detección de una
proteína específica en
algún tejido.
WESTERN BLOT
* Análisis de α-SMA y
COL1.
* Análisis de TLR4,
MyD88, TAK1, p65,
p-p65, IκBα, p-IκBα
and IKKβ.
Para qué Fundamento
RESULTS
Untreated cells were used as control.
RESULTS
RESULTS
RESULTS
LPS can cause the expression of IL-6,
IL-1β and TNF-α and the expression of
collagen I, indicating that LPS could
induce inflammation and fibrosis in
LX2 cells.
PHI could inhibit the activation of LX2
cells and the expression of fibrotic
cytokines by inhibiting the
proinflammatory reaction induced by
LPS in the TLR4 / MyD88 / NF-κB
signaling pathway.
DISCUSSION
Author Concept
Liu et al.; Paik et al.
LPS can not only stimulate LX2 cells to
secrete inflammatory cytokines, but
also activate LX2 cells to express
fibrotic cytokines of α-SMA and COL1.
Jaeyeon Jung
It has been reported that inhibition of
NF-κB signaling pathway played an
important role in reducing liver fibrosis
reaction.
Chen et al.; Liu et al. LPS induced LX2 cells inflammation
model was successfully established
with upregulation of IL-1β, IL-6 and
TNF-α levels.
CONCLUSIONS
Some substances of herbal origin such
as phillygenin can produce effects on
inflammatory processes by interfering in
a specific signaling pathway and in this
way cause a therapeutic response on
some tissue that could be compromised.
LPS are one of the responsible for the
production of proinflammatory
cytokines in this case by the activation of
the TLR4 / MyD88 / NF-κB pathway
where it eventually causes liver tissue
damage.
Seminario biologia molecular jazmin gomez

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Seminario biologia molecular jazmin gomez

  • 1. Jazmin Gómez Olarte Tercer semestre 2020-1 Facultad de medicina, Universidad Pontificia Bolivariana
  • 2. INTRODUCTION Phillygenin is a component extracted from Forsythiae Fructus. Forsythia Suspensa Forsythiae Fructus Phillygenin * Detoxify * Therapeutic effects
  • 3. INTRODUCTION TLR4/MYD88/NF-kB pathway LPS: Lipopolysaccharide TLR: Toll-like receptors PHI could inhibit the proinflammatory reaction induced by LPS, in addition it could inhibit protein expression in the TLR4 / MyD88 / NF-κB signal pathway.
  • 4. OBJECTIVE Investigate if Phillygenin (PHI) has a therapeutic effect on liver fibrosis caused by inflammation.
  • 5. MATERIALES Y MÉTODOS Para qué Para cultivo de las células LX2 (línea de las células estrelladas hepáticas del humano). CULTIVO CELULAR Fundamento Proceso mediante el que las células pueden cultivarse en condiciones controladas.
  • 6. Fundamento Para qué ELISA Enzyme-Linked ImmunoSorbent Assay MATERIALES Y MÉTODOS Detección de los factores inflamatorios IL-6, IL-1β y TNF-α. *Prueba para determinar [Antígeno] o [Anticuerpo] con uno de ellos en fase sólida y el otro en solución. * Conjugación enzima-Ac que genera un producto detectable.
  • 7. MATERIALES Y MÉTODOS Fundamento * Retrotranscripción de una hebra de ARN en ADN. * Uso de transcriptasa inversa. RT-qPCR Reacción en cadena de la polimerasa con transcriptasa inversa Para qué Determinación de la expresión de colágeno I y α-SMA (secreción de actina del músculo liso α) y para verificación del mecanismo de PHI en la ruta de señal TLR4 / MyD88 / NF-κB
  • 8. MATERIALES Y MÉTODOS Usado para la detección de una proteína específica en algún tejido. WESTERN BLOT * Análisis de α-SMA y COL1. * Análisis de TLR4, MyD88, TAK1, p65, p-p65, IκBα, p-IκBα and IKKβ. Para qué Fundamento
  • 9. RESULTS Untreated cells were used as control.
  • 12. RESULTS LPS can cause the expression of IL-6, IL-1β and TNF-α and the expression of collagen I, indicating that LPS could induce inflammation and fibrosis in LX2 cells. PHI could inhibit the activation of LX2 cells and the expression of fibrotic cytokines by inhibiting the proinflammatory reaction induced by LPS in the TLR4 / MyD88 / NF-κB signaling pathway.
  • 13. DISCUSSION Author Concept Liu et al.; Paik et al. LPS can not only stimulate LX2 cells to secrete inflammatory cytokines, but also activate LX2 cells to express fibrotic cytokines of α-SMA and COL1. Jaeyeon Jung It has been reported that inhibition of NF-κB signaling pathway played an important role in reducing liver fibrosis reaction. Chen et al.; Liu et al. LPS induced LX2 cells inflammation model was successfully established with upregulation of IL-1β, IL-6 and TNF-α levels.
  • 14. CONCLUSIONS Some substances of herbal origin such as phillygenin can produce effects on inflammatory processes by interfering in a specific signaling pathway and in this way cause a therapeutic response on some tissue that could be compromised. LPS are one of the responsible for the production of proinflammatory cytokines in this case by the activation of the TLR4 / MyD88 / NF-κB pathway where it eventually causes liver tissue damage.