Teratogenic study of phenobarbital and levamisole on mouse fetus liver tissue using biospectroscopy.
Biospectroscopic investigations have attracted attention of both the clinicians and basic sciences researchers in recent years. Scientists are discovering new areas for FTIR biospectroscopy applications in medicine. The aim of this study was to measure the possibility of FTIR-MSP application for the recognition and detection of fetus abnormalities after exposure of pregnant mouse to phenobarbital (PB) and levamisole (LEV) alone or in combination. PB is one of the most widely used antiepileptic drugs (AEDs), with sedative and hypnotic effects. When used by pregnant women, it is known to be a teratogenic agent. LEV is an antihelminthic drug with some applications in immune-deficiency as well as colon cancer therapy. Four groups of ten pregnant mice were selected for the experiments as follows: one control group received only standard diet, one group was injected with 120mg/kg of BP, one group was injected with 10mg/kg of LEV, and the last group was treated simultaneously with both BP and LEV at the above mentioned doses. Drugs administration was performed on gestation day 9 and fetuses were dissected on pregnancy day 15. Each dissected fetus was fixed, dehydrated and embedded in paraffin. Sections of liver (10μm) were prepared from control and treated groups by microtome and deparaffinized with xylene. The spectra were taken by FTIR-MSP in the region of 4000-400cm(-1). All the spectra were normalized based on amide II band (1545cm(-1)) after baseline correction of the entire spectrum, followed by classification using PCA, ANN and SVM. Both morphological and spectral changes were shown in the treated fetuses as compared to the fetuses in the control group. While cleft palate and C-R elongation were seen in PB injected fetuses, developmental retardation was mostly seen in the LEV injected group. Biospectroscopy revealed that both drugs mainly affected the cellular lipids and proteins, with LEV causing more changes in amide I and lipid regions than PB. Application of PCA, ANN and SVM methods were able to successfully classify these FTIR spectroscopic data and discriminate between control and treated groups of fetuses, making it a new potential tool for drugs teratogenic investigations.
Occupational Health Research Center, School of Public Health, Iran University of Medical Sciences, Tehran, Iran; Department of Occupational Health, School of Public Health, Iran University of Medical Sciences, Tehran, Iran.
Azadeh
A
Ashtarinezhad
Young Researchers and Elite club, Central Tehran Branch, Islamic Azad University, Tehran, Iran.
Ataollah
A
Panahyab
Data Mining Department, Tosan Intelligent Data Miners Co., Tehran, Iran.
Shahrzad
S
Shaterzadeh-Oskouei
Data Mining Department, Tosan Intelligent Data Miners Co., Tehran, Iran.
Hessam
H
Khoshniat
Department of Toxicology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Baharak
B
Mohamadzadehasl
Department of Toxicology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Pharmaceutical Sciences Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Electronic address: f.shirazi@sbmu.ac.ir.
Farshad H
FH
Shirazi
D000014:Abnormalities, Drug-Induced / Q000473:pathology; D000818:Animals; D000871:Anthelmintics / Q000633:toxicity*; D000927:Anticonvulsants / Q000633:toxicity*; D004347:Drug Interactions; D005260:Female; D005333:Fetus / Q000473:pathology; D007978:Levamisole / Q000633:toxicity*; D008099:Liver / Q000737:chemistry / Q000196:embryology / Q000473:pathology*; D051379:Mice; D009152:Mutagenicity Tests; D016571:Neural Networks, Computer; D010634:Phenobarbital / Q000633:toxicity*; D011247:Pregnancy; D025341:Principal Component Analysis; D017550:Spectroscopy, Fourier Transform Infrared; D060388:Support Vector Machine; D013723:Teratogens / Q000633:toxicity*
D000871:Anthelmintics; D000927:Anticonvulsants; D013723:Teratogens; D007978:Levamisole; D010634:Phenobarbital
Biospectroscopy; Fetus abnormalities; Levamisole; Phenobarbital; Teratogenic agent