article about cell swelling

1. Presented by: Santiago Del Castillo Alvarez
Universidad Pontificia Bolivariana
2020-1

2. Introduction
EGFR
ERK
PKC
● Membrane
receptor for
EGF.
● Tyrosine
Kinase
● Modulates
multiple
biological
processes
● Signaling
via that
stimulates
mitosis.
● Same as
MAPK
● Kinase family
involved in cell
growth and
differentiation,
hormone
secretion, and
apoptosis.
● Calcium
dependent

3. Introduction
EGFR
ERK
PKCδ

4. Overall Objective
The study aimed to clarify the involvement of PKC in ammonia-
induced EGFR transactivation and subsequent ERK activation
to help provide further insight into the pathogenesis of HE and
guide the development of new treatment strategies.

5. Materials and
Methods
I. Nerve cells were extracted from 1 day old
rats.
II. Harvested cells were cultured in eagle
medium with humidified atmosphere.
III. Nine days later, microglia and
oligodendrocyte progenitors were removed
by shaking 2 to 3 times.
IV. Obtention of preparations with 95% to 99%
astrocytes.
Primary astrocyte culture

6. Materials and
Methods
SDS-PAGE Western Blot
I. Addition of SDS to
proteins denatures
them and confers
negative charge.
II. Addition of proteins
to PAGE with electric
current.
III. migration of proteins
by molecular mass to
the anode.
I. Incubation with primary
antibodies against ERK
and P-ERK for 2 hours.
II. Incubation with
HP-secondary antibodies.
III. visualization of the
bands.
IV. Quantifying optical
density of the bands.

7. Materials and
Methods
I. Whole-cell lysates with proteins incubated 12 hours
with Anti- EGFR/ Src to form immune complex.
II. addition of agarose, incubation 2 more hours.
III. centrifugation and boiling.
IV. collect supernatant.
V. Western blot.

8. Results

9. Results

10. Results

11. Results

12. Results

13. Discussion
Saxena et al. 2017; T. Wang et al. 2018;
Wang et al. 2017; Yi et al. 2019
PKC activation leads to the phosphorylation
and activation of downstream signaling
proteins that control a variety of bio- logical
responses, including cell cycle regulation,
autophagy, apoptosis, and bipolar disorder
Hsieh et al. 2008; Huang et al. 2013; Yang
et al. 2009
PKC-dependent Src activation is well
established in many biological processes
Buzanska et al. 2000 In addition, it was previously documented
that a pathological concentration of
ammonia can trigger a transient increase in
PKC activity, contributing to delayed
apoptosis of C6 glioma cells.

14. Conclusions
● Ammonia increased blood levels causes cell swelling by
transactivation of EGFR which is normally activated by
EGF stimulating cell growth And mitosis.
● BIM and Rottlerin could be used as new
pharmacological treatment and/or prevention of Brain
edema.

15. Brain edema
Is a
Pathological increase in the
waterpolo content of the brain
Can be caused by
Cancer
Trauma
Hyperammonemia
Produces
Neuropsychiatric disorders and
brain herniation
Its related to hepatic
diseases