The study of Slc25a3 gene effect in apoptosis of cardiomyocytes in SD Rats by doxorubicin and the protective mechanism using by curcumin.- Dr Sudeep KC

1. PostGraduation Student: Name: Sudeep K.C.
Faculty (major): Internal Medicine (Cardiology)
Professior name ： Lu JunKun
Thesis defense date ： June 2015
The study of Slc25a3 gene effect in apoptosis of
cardiomyocytes in SD Rats by doxorubicin and the
protective mechanism using by curcumin.

2. The study of Slc25a3 gene effect in apoptosis of cardiomyocytes in
SD Rats by doxorubicin and the protective mechanism using by
curcumin.

3. 1. Preface
2. Aims & Objectives
3. Methods
4. Result
5. Discussion
6. Conclusion

4. PREFACE
Doxorubicin causes acute myocardial injury, leading to
myocardial interstitial edema, myocardial fibers derangement,
swelling and heart volume to be increased significantly .
Doxorubicin cardiomyopathy mechanism:
Existing relevant ultrastructural studies have found that anthracycline-
induced myocardial cell injury two main forms, namely myocardial
fiber necrosis and degeneration of the myocardium, these changes
can lead to progressive myocardial necrosis, electron microscopy
shows myocardial muscle fibers disorder, muscle dearray,
mitochondrial hyperplasia, swelling, deformity, and reduce cristae.
Studies have also indicated that doxorubicin may lead to non-species-
dependent myocardial injury . Although the exact mechanism of
doxorubicin-induced cardiotoxicity still not clear, but most studies
have shown that oxidative stress may play the most critical role, which
is due to the chemical structure of doxorubicin in the metabolic
process to produce reactive oxygen species

5.  The purpose of this experiment is to study the
Protective effect of curcumin against doxorubicin
induced Myocardial cell injury on SD rats and to
explore the expression of Pic Protein and Slc25a3 gene
inhibition on Doxorubicin induced Myocardial injury
under Curcumin intervention, depending upon the
different concentration and duaration of use of
curcumin.
 To study the protective effect of Curcumin against
Doxorubicin induced myocardial injury on SD rats by
inhibiting expression of Slc25a3 gene.
Aims&Objectives

6. Methods
 Normal healthy adult SD rats (male and female, 180-
220g) were used to establish doxorubicin-induced
myocardial injury animal models, randomly divided
into three groups: control group, doxorubicin group,
curcumin with doxorubicin group.

7. Control group : was injected 0.9% sodium
chloride injection (2.5ml/kg).
Doxorubicin group :was injected diluents of doxorubicin
i.e. 2.5mg/kg, by 0.9% sodium chloride injection .
 Curcumin with doxorubicin group was injected
diluents of doxorubicin i.e. 2.5mg/kg, by 0.9% sodium
chloride injection with curcumin (30mg/kg).
All groups were injected via rat tail vein once every
week following for 6 weeks .

8. After eleven weeks, all rats were sacrificed using spinal
cord transection method and heart tissues were taken
for the next experiment.
Flow cytometry is used to test the SD rat
cardiomyocytes transferred level; Application of Real-
time PCR technology is used to detect Slc25a3 gene
expression.

9. Using SPSS 16.0 software, the results were expressed
as mean ± standard deviation (Mean± SD). Data
between the two groups were compared using t test,
among groups were compared using ANOVA. With P
<0.05 was considered statistically significant

10. Result
Detection of cardiomyocytes apoptosis in SD rats by flow
cytometry
In this study, flow cytometry is used in animal model to detect
myocardial apoptosis.
The results showed that apoptotic rate was significantly higher
in doxorubicin injury group than curcumin intervention group
(Figure), the differences were statistically significant (p <0.05).
Figure - Animal models of myocardial apoptosis

11. Result
Level of Expression of PiC after curcumin on adriamycin-induced myocardial
cell injury on SD Rats:
The expression of Slc25a3 mRNA was significantly higher in
doxorubicin injury group than that in control group and curcumin
intervention group, while the expression of Slc25a3 mRNA is
significantly lower in curcumin intervention group than that in
control group (p <0.01, p <0.05).
Figure 16 expression of PiC after curcumin intervention in adriamycin-induced
myocardial cell injury in SD rats

12. Discussion
Doxorubicin is broadly used antitumor drug. The study
shows patients using long duration of doxorubicin cause
cardiac toxicity, till now the exact mechanism is not clear
but many research and experiments have done and shown
that oxidative stress plays a critical role due to chemical
structure of doxorubicin to produce reactive oxygen species
and cause myocardial injury and mitochondrial sub cellular
injury.

13. Curcumin a family of Zingiberacae, is a plant used for spice, coloring and
a major source of curry powder in country like India and has a long
traditional value in medicine in country like China.
Many studies shown curcumin to be inhibitor of tumor initiation in vivo.
It also showed cardioprotective effect by suppression of myocardial
protein expression of inducible nitric oxide synthase and the catalytic
subunit of nicotinamide adenine dinucleotide phosphate reduced
oxidate and significantly decrease myocardial endoplasmic reticulum
stress signaling proteins.
Discussion

14. Mitochondrial is a protein that in humans encoded by Slc25a3gene. The
protein encoded by this gene catalyzes the transport of phosphate into the
mitochondrial matrix, either by protein co-transport or in exchange for
hydroxyl ions.
The experiment here is to study the Protective effect of curcumin against
doxorubicin induced Myocardial cell injury on SD rats and to explore the
expression of Pic Protein and Slc25a3 gene inhibition on Doxorubicin
induced Myocardial injury under Curcumin intervention, depending upon
the different concentration and duaration of use of curcumin.
We studies the Slc25a3 gene effect in apoptosis of cardiac injury in SD rats
by establishing protective mechanism of curcumin against doxorubicin
injuced myocardial injury.
Discussion

15. 结论（ Conclusion ）
• 1. Doxorubicin intervention in SD rats myocardial cells can
induce myocardial cells apoptosis, curcumin can significantly
reduce the doxorubicin-induced myocardial cells apoptosis.
• 2. The expression of PiC and Slc25a3 gene on doxorubicin-
induced myocardial injury in rats have an important role. The
Slc25a3 gene up-regulation is closely related to doxorubicin
myocardial injury. Curcumin down regulates Slc25a3 gene
expression, thus reducing doxorubicin induced myocardial cell
damage in rats.

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