Inhibitory Effects of Isoliensinine on Bleomycin-Induced Pulmonary Fibrosis in Mice

Abstract

The effects of isoliensinine (IL), a bisbenzylisoquinoline alkaloid extracted from the Chinese traditional medicine seed embryo of Nelumbo nucifera Gaertn., on bleomycin (BLM)-induced pulmonary fibrosis in mice were investigated. Seventy-two male Kungming mice were divided randomly into eight groups as BLM-IL10, BLM-IL20, BLM-IL40, BLM-Sal, Sal-IL10, Sal-IL20, Sal-IL40 and Sal-Sal groups. BLM (0.1 mg in 0.05 ml saline per animal, once) or saline (0.05 ml per animal, once) was applied intratracheally, and IL (10, 20, 40 mg/kg) or saline was administered orally 3 times per day in the appropriate groups. Animals were sacrificed 14 days after intratracheal treatment. Lung tissue and serum superoxide dismutase (SOD) activity and malondialdehyde (MDA) levels, tumor necrosis factor α (TNF-α) and transforming growth factor beta 1 (TGF-β₁) were determined by biochemical measurements and immunohistochemistry. BLM treatment resulted in a significant increase of the hydroxyproline content and an obvious lung histological injury as compared to the Sal-Sal group. Administration of IL remarkably suppressed the increase in hydroxyproline content and abated the lung histological injury induced by BLM. There was a decrease in SOD activity and an increase in MDA level in lung tissue and serum in the BLM-Sal group (p < 0.01, p < 0.01, vs. Sal-Sal group, respectively). And IL could obviously enhance the SOD activity and decrease the MDA level in a concentration-dependent manner. Moreover, IL also significantly inhibited the overexpression of TNF-α and TGF-β₁ induced by BLM. These results indicated that IL possessed a significant inhibitory effect on BLM-induced pulmonary fibrosis, probably due to its antioxidant and/or anti-inflammatory activities and inhibitory overexpressing TNF-α and TGF-β₁ induced by BLM.

Abbreviations

bFGF:Basic fibroblast growth factor

BLM:Bleomycin

DAB:Diaminobenzidine tetrahydrochloride

IL:Isoliensinine

IT:Intratracheal

MDA:Malondialdehyde

NBT:Nitroblue tetrazolium

PBS:Phosphate buffer solution

PDGF:Platelet derived growth factor

PF:Pulmonary fibrosis

PO:per os, orally

ROS:Reactive oxygen species

SABC:Strept-avidin-biotin-peroxidase-complex

Sal:Saline

SOD:Superoxide dismutase

TGF-β:Transforming growth factor beta

TNF-α:Tumor necrosis factor α

Key words

Isoliensinine - bisbenzylisoquinoline - bleomycin - pulmonary fibrosis - SOD - MDA - TNF-α - TGF-β₁

References

• 1 Taooka Y, Maeda A, Hiyama K, Ishioka S, Yamakido M. Effects of neutrophils elastase inhibitor on bleomycin-induced pulmonary fibrosis in mice.  Am J Respir Crit Care Med. 1997;  156 260-5
• 2 Mason R J, Schwarz M I, Hunninghake G W, Musson A R. Pharmacological therapy for idiopathic pulmonary fibrosis: past, present, and future.  Am J Respir Crit Care Med. 1999;  160 1771-7
• 3 Wang H, Yamaya M, Okinaga S, Jia Y, Kamanaka M, Takahashi H. et al . Bilirubin ameliorates bleomycin-induced pulmonary fibrosis in rats.  Am J Respir Crit Care Med. 2002;  165 406-11
• 4 Feldman M, Bondeson J, Brennan F M, Foxwell B M, Maini R N. The rational for current boom in anti-TNF alpha treatment.  Is there an effective means to define therapeutic targets for drugs that provide all benefits on anti-TNF alpha and minimize hazards? Ann Rheum Dis. 1999;  58 (. 1) 127-31
• 5 Wang J L, Hu X M, Yin W H, Cai H. Alkaloids of Plumula nelumbinis .  [in chinese] Zhongguo Zhong Yao Zhi (Chin J Clin Mat Med). 1991;  16 573-5, 703
• 6 Zhang F, Wang J L, Qian J Q. Effects of isoliensinine on experimental arrhythmia and myocardium action potential of guinea pig.  Chin Tradit Herb Drugs. 2000;  31 53-5
• 7 Feng X L, Yu X, Xiao J H, Wang J L. Effect of isoliensinine on cardiovascular function and experimental left ventricular hypertrophy.  J Huazhong Univ Sci Tech [Health Sci]. 2002;  31 608-11
• 8 Qian J Q. Cardiovascular pharmacological effects of bisbenzylisoquinoline alkaloid derivatives.  Acta Pharmacol Sin. 2002;  23 1086-92
• 9 Selman M, Montano M, Ramos C, Barrios R, Perez-Tamayo R. Experimental pulmonary fibrosis induced by paraquat plus oxygen in rats: a morphologic and biochemical sequential study.  Exp Mol Pathol. 1989;  50 147-66
• 10 Adamson I Y, Bowden D H, Cote M G, Witschi H. Lung injury induced by butylated hydroxytoluene: cytodynamic and biochemical studies in mice.  Lab Invest. 1977;  36 26-32
• 11 Luna M A, Bedrossian C W, Lichtiger B, Salem P A. Interstitial pneumonitis associated with bleomycin therapy.  Am J Clin Pathol. 1972;  58 501-10
• 12 Jeremy T A, Monica A S. Growth factors in idiopathic pulmonary fibrosis: relative roles.  Respir Res. 2002;  13 1-9
• 13 Lossos I S, Izbicki G, Or R, Goldstein R H, Breuer R. The effect of suramin on bleomycin-induced lung injury.  Life Sciences. 2000;  67 2873-81
• 14 Woessner J F. The determination of hydroxyproline in tissue and protein samples containing small proportions of this amino acid.  Arch Biochem Biophys. 1961;  93 440-7
• 15 Sun Y, Oberley L W, Li Y. A simple method for clinical assay of superoxide dismutase.  Clin Chem. 1988;  34 497-500
• 16 Ohkawa H. Assay for lipid peroxides in animal tissues by thiobarbituic acid reaction.  Anal Biochem. 1979;  95 351
• 17 Jules-Elysee K, White A. Bleomycin-induced pulmonary toxicity.  Clin Chest Med. 1990;  11 1-20
• 18 Ozyurt H, Sogut S, Yldrm Z, Kart L, Iraz M, Armutcu F. et al . Inhibitory effects of caffeic acid phenethyl ester on bleomycine-induced lung fibrosis in rats.  Clinica Chimica Acta. 2004;  339 65-75
• 19 Giri S N, Hyde D N, Hollinger M A. Effect of antibody to transforming growth factor beta on bleomycin induced accumulation of lung collagen in mice.  Thorax. 1993;  48 959-66
• 20 Raghow B, Irish P, Kang A H. Coordinate regulation of transforming factor beta gene expression and cell proliferation in hamster lungs undergoing bleomycin-induced pulmonary fibrosis.  J Clin Invest. 1989;  84 1836-42