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Study on Cellular Damage of Human L-02 Hepatocyte Induced by Roundup (41% Glyphosate)

Author: WangFei
Tutor: ZhongCaiGao
School: Central South University
Course: Health Toxicology
Keywords: Roundup Liver cells Oxidative damage Mitochondrial damage Apoptosis
CLC: R363
Type: Master's thesis
Year: 2008
Downloads: 61
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Abstract

Objective: To investigate Roundup (41% glyphosate) human L-02 liver cells injury and its possible mechanism. Methods: In vitro tests L-02 liver cells were cells by MTT assay different concentrations (in vitro test) Roundup L-02 liver cell viability, select the cell survival rate was 20% -80% concentration for subsequent experiments. Set 5 Roundup treated with 60mg / L, 90mg / L, and 120mg / L, 150mg / L, 180 mg / L and a negative control group (PBS), Roundup treating cells for 24h. L-02 liver cells morphological changes Kyi the mother Sa staining method light microscope combined with transmission electron microscopy; Roundup of the L-02 liver cell membrane permeability affect and cell toxicity of aspartate aminotransferase (AST), alanine aminotransferase (ALT) activity levels combined with trypan blue exclusion staining method to evaluate; Roundup induced L-02 liver cells from oxidative damage degree of selection of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH ) content and other indicators evaluation; to examine mitochondrial damage mitochondrial transmembrane potential (△ Ψm); DNA bands (DNA-Ladder) to detect hepatocellular DNA damage; to AnnexinV-FITC/PI the complex staining was measured apoptosis necrosis; Western Blotting detection control group and 90mg / L cytochrome C (Cyt C) and apoptosis-inducing factor (AIF) expression levels. Results: 1. Within 60 ~ 180mg / L concentration range, significantly reduced (P <0.05) caused by L-02 liver cell survival, there is a negative correlation between the concentration and cell viability (r = -0.974 ). 2 climbing film Kyi mother Sa staining L-02 liver cells in the light microscope, observed Roundup treated L-02 liver cell shrinkage or swelling, enlarged morphology by tile-like contraction of the circular, cell gap cell rupture, chromatin condensation, density and reduce the number of adherent cells. Electron microscopy, the treated cells can be found in surface microvilli disappear, incomplete cell membrane structure, nuclear fragmentation or swelling, mitochondrial vacuolar degeneration and apoptosis or necrosis performance. Roundup handling L-02 liver cells, live cells by trypan blue blue dye ratio increased (P <0.05); cell culture supernatant of ALT activity increased (P <0.05); starting from 90mg / L group AST activity increase (P <0.05); Roundup treated cells increased the content of MDA, SOD activity decreased GSH content decreased (P <0.05); Roundup lead 120 ~~ 180mg / L of cell Na ~-K ~ ATP enzyme The activity is decreased (P <0.05). 4. Roundup treatment can induce DNA strand breaks, to deal with the concentration of 150mg / L, 180 mg / L, the degree of DNA damage serious; 90mg / L group Cyt C and AIF expression levels higher than that of the control group (P <0.05); 90mg L-02 cells / L group began treatment Roundup mitochondrial membrane potential was significantly lower (P <0.05); the Roundup treated significantly induced liver L-02 cells to apoptosis and necrosis (P <0.05), with the processing of agricultural higher degrees higher proportion of apoptosis and necrosis, but in the 180mg / L the proportion of apoptotic decline. Conclusion: Roundup in the range of 60mg / L ~ 180mg / L can cause a decline in L-02 liver cell viability, increased membrane permeability, inhibition of cell ion transport, induce DNA damage, mitochondrial membrane potential, Cyt C and AIF leaks and other apoptotic factors, cell apoptosis and necrosis of liver cells with visible damage; damage mechanism Roundup may lead to oxidative damage of the liver cells, mitochondrial collapse pathway.

Study on Cellular Damage of Human L-02 Hepatocyte Induced by Roundup (41% Glyphosate)

Abstract

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