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Study on Mechanisms of Testis Oxidative Injury and Apoptosis in Freshwater Crab (Sinopotamon henanense) Induced by Cadmium

Author: XuTuan
Tutor: WangLan
School: Shanxi University
Course: Zoology
Keywords: Henanense Testis Oxidative damage Apoptosis Cadmium
CLC: X174
Type: Master's thesis
Year: 2011
Downloads: 18
Quote: 0
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Abstract


In this paper, the acute toxicity test methods, cadmium (Cd2) the testis the of henanense (Sinopotamon henanense) the impact of oxidative damage and apoptosis. The experiment set of five the CdCl2 treatment groups (7.25,14.50,29.00,58.00,116.00 mg / L) and a control group, time 1,3,5 and 7d. In order to study the Cd2 impact on testis oxidative stress, were detected by superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT) activity and malondialdehyde aldehyde (MDA) content. In order to explore Cd2 whether in testis oxidation damage after further cause spermatogenic cell apoptosis, respectively, using hematoxylin - eosin (HE) staining, microscopic and submicroscopic technology and agarose gel electrophoresis, from cell morphology spermatogenic cell apoptosis detection and biochemical level. Conducted a preliminary exploration of Cd2 induced the testes apoptosis mechanism. Different concentrations of Cd2 7d, respectively, determined by the content of hydrogen peroxide (H2O2), half caspases -3 (Caspase-3), half caspases -9 (Caspase-9), and cytochrome c oxidase ( CCO) activity. The results showed that: (1) in the testis SOD, GPx, CAT activity presented with the the Cd2 concentration increase and treatment time trend first and then decreased. Low concentrations of Cd2 significantly induced increase in antioxidant enzyme activity; When the Cd2 concentration increases processing time, the enzyme activity decreased below the level of the control group. MDA content is presented with the the Cd2 concentration increase and treatment time gradually increased. The suppression of the antioxidant enzyme activity and MDA content increased, indicating that testis serious oxidative damage. (2) HE staining of spermatogenic cells in the seminiferous epithelium of the control group arranged orderly form of structural integrity, no shedding of spermatogenic cells in the lumen. With the Cd2 concentration increased and the time of the extension of a disordered arrangement of spermatogenic cells, spermatogenic cells with reduced sperm count, germinal layer vacuoles histopathological changes. Fluorescence microscopy revealed apoptotic spermatogenic cells gradually increased. Transmission electron microscopy (TEM) observation that spermatogenic cells showed typical characteristics of apoptosis, specific performance for membrane rupture, mitochondrial vacuolization and nuclear membrane shrinkage. Agarose gel apoptotic cells by specific DNA ladder. (3) processing 7d With Cd2 concentration increase, H202 content, Caspase-3 and Caspase-9 vitality gradually increasing trend, a gradual downward trend CCO vitality. Show that the the cadmium induced spermatogenic cells produce a large number of reactive oxygen species, destruction of the mitochondrial electron transport chain, Caspase-9-dependent mitochondrial pathway of apoptosis of spermatogenic conclusion: (1) low concentrations the Cd2 able to activate Henan the testis tissue Sinopotamon the antioxidant enzyme activity, to clear Cd2 induced reactive oxygen radicals; high concentrations of Cd2 inhibition of antioxidant enzyme activity in the testis redox balance is broken, the organization increased active oxygen membrane lipid peroxidation, eventually leading to the testes in severe oxidative stress status. (2) acute Cd2 exposure can lead to apoptosis henanense reproductive toxicity and germ cells, and cadmium toxicity of a cumulative and incremental process. Cadmium-induced apoptosis of spermatogenic cells process, the structure and function of mitochondrial damage and apoptosis Caspase-9 and Caspase-3 is activated, cadmium induced testis apoptosis mediated Caspase-9 mediated mitochondrial apoptotic pathway induced. (3) MDA content can be used as the body subjected to of Cd2 oxidative stress biomarkers, can provide a scientific basis for water quality monitoring, aquatic health, breeding.

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