EFFECTS OF ERDOSTEINE ON INFLAMMATION AND FIBROSIS
IN RATS WITH PULMONARY FIBROSIS INDUCED BY
BLEOMYCIN
Ersin Şükrü Erden1, Gamze Kırkıl2, Figen Deveci2, Nevin İlhan3, Bengü Çobanoğlu4, Teyfik Turgut2,
Mehmet Hamdi Muz2
Erciş State Hospital, Van1
Firat University, Faculty of Medicine, Department of Chest Disease2, Elazig
Firat University, Faculty of Medicine, Department of Biochemistry3, Elazig
Firat University, Faculty of Medicine, Department of Pathology4 ,Elazig
Introduction and aim
Idiopathic pulmonary fibrosis (IPF) is an interstitial pulmonary disease that has a poor prognosis with current treatment modalities
There are 2 phases in the pathophysiology of experimental lung fibrosis induced by BLM, an antineoplastic agent,
Early inflammatory phase; infiltration of macrophage, neutrophil, and lymphocytes in interstitium and alveolus
Second phase; fibrosis
It is shown that erdostein, one of mucolytic agent,
Supressed the inactivation of 1-antitrypsin in human induced by smoking, and increased the airway secretions of mice and rabbits.
Aims1. To obtain the data that can contribute
to clarify the pathogenesis of IPF in the experimental lung fibrosis model induced by BLM
2. To determine the effect of erdosteine on acute inflammatory changes and fibrosis, contribution of antioxidant therapy to IPF treatment
Material and method
45 Wistar male rats classified into 3 groups
5 of them on 0th day 5 of them on 14th ay 5 of them on 29th day were sacrificed
-2th day 0th day 14th day 29th day
NaHCO3 (10 mg/kg/d, 1x1, po)
it 0.2 mg PBS
Group 1 (n=15)
NaHCO3 (10 mg/kg/d, 1x1, po)
it BLM 7.5u/kg
Group 2 (n=15)
Group 3 (n=15)
it BLM 7.5u/kg
Erdostein (10 mg/kg/gd1x1, po)
29th day
29th day
14th day
14th day
0th day
0th day
-2th day
-2th day
Performing BAL BAL was performed by flushing the
airways with 1 ml PBS through the tracheal cannula five times. Approximately 4.5 ml (90%) aliquot was withdrawn
MDA, MIP-1α, MIP-2 levels were measured in BALF
Cell classification of BALF
After cytospin procedure, the cell smear was stained with May-Grünwald-Giemsa
White cell count and NG were measured by counting 200 cells under light microscopy.
Histopathologic Analysis of the Lung
After performing BAL, left lung were fixed in 10% of formaldehyde, and the tissues were cut
Right lung was used to measure the hydroxyproline levels
Statistical Analysis
To compare the data of more than two groups,the non-parametric Kruskal-Wallis test was used.
Comparisons between groups were performed using the Mann-Whitney U test
RESULTS
lenfosit nötrofil
BA
L h
ücre
(%
)
8
7
6
5
4
3
2
Kontrol
Bleomisin
Erdostein
BA
L ce
ll (%
)
neutrophil lymphocyte
control
Bleomycin
Erdostein
BAL cell classification on day O
BAL cell classification on day 14th
lenfositnötrofil
BA
L h
ücre
(%
)
30
20
10
0
Kontrol
Bleomisin
Erdostein
Neutrophil
CxBLM p=0.008CxE p=0.007BLMxE p=0.008
Lymphocyte
CxBLM p=0.009
neutrophillymphocyte
BA
L ce
ll (%
)
control
Bleomycin
Erdostein
BAL cell classification on day 29th
lenfositnötrofil
BA
L h
ücre
(%
)
20
10
0
Kontrol
Bleomisin
Erdostein
Neutrophil
CxBLM p=0.009CxE p=0.012BLMxE p=0.009
Lymphocyte
CxBLM p=0.027BA
L ce
ll (%
)
neutrophil lymphocyte
control
Bleomycin
Erdostein
BAL MIP-1 levels of all groups on days 0, 14th and 29th
29. gün14. gün 0. gün
BA
L M
İP-1
alf
a (p
g/m
l)14
12
10
8
6
4
2
Kontrol
Bleomisin
Erdostein
BA
L M
IP-1
(p
g/m
l)
Oth day 14th day 29th day
control
Bleomycin
Erdostein
14th day
CxBLM p=0.009CxE p=0.009BLMxE p=0.016
29th day
CxBLM p=0.009BLMxE p=0.009
BAL MIP-2 levels of all groups at 0th,14 th,29 th days
29. gün14. gün0. gün
BA
L M
İP-2
(pg
/ml)
500
400
300
200
Kontrol
Bleomisin
Erdostein
14th dayOth day 29th day
control
Bleomycin
Erdostein
14th day
CxBLM p=0.009BLMxE p=0.009
29th day
CxBLM p=0.009BLMxE p=0.016
BAL MDA levels of all groups at 0th,14th,29th days
29. gün14. gün0. gün
BA
L M
DA
(nm
ol/m
l)
2,2
2,0
1,8
1,6
1,4
1,2
Kontrol
Bleomisin
Erdostein
14th dayCxBLM p=0.009CxE p=0.016BLMxE p=0.009
29th day
CxBLM p=0.012BLMxE p=0.009
Oth day 14th day 29th day
control
Bleomycin
Erdostein
Tissue levels of OH-P and fibrosis scores of all groups
Control group(n=15)
BLM group(n=15)
Erdostein group(n=15)
0th day
OH-P (mg/gr) 9.73±1.86 10.63±2.44 10.42±2.03
Fibrosis 0.40±0.54 0.40±0.54 0.40±0.54
14th day
OH-P (mg/gr dry tissue
10.52±1.65 15.90±1.99 11.06 ±1.94
Fibrosis 0.20±0.44 1.60±0.89 1.40±0.54
29th day
OH-P (mg/gr dry tissue)
10.12±2.40 19.33±1.15 11.45±0.94
Fibrosis 0.20±0.44 3.00±0.00 1.60±0.54
The histopathological views of lung tissues of control group (A), BLM group (B), and
erdostein group (C)
A) B)
(C)
Grade 3 fibrosis
Decrease in fibrosis
The inflammatory cell infiltration in BALF of control group (A), BLM group (B), and
Erdostein group (C)
A) B)
(C)
In conclusion
Erdostein may prevent the acute lung inflammation and fibrosis induced by BLM in rats
This protective effect of erdostein may be due to supression the accumulation of neutrophils, inhibition of lipid peroxydation, chemokine production, and release.
Our results may suggest that erdostein can be a new pharmacological agent for IPF therapy,
Erdostein can be used as a proflactic agent during the use of antineoplastic agents especially in early phases