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 Approximately 4.5 ml (90%) aliquot was withdrawn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