The Mechanism Of Microcystin-Induced Oxidative Damage Of Rat Liver Cells And The Protective Effect Of Vitamin B Group & Vitamin C

Objectives:To study the mechanism of microcystin-induced oxidative damage in vitro with molecular biology technologies, and the effect of vitamin B group & antioxidant nutrients protecting liver cells from microcystin injury, which would help us find an effective way to protect liver and prevent liver cancer.Methods:1,Purchase BRL cells, initiate the testing after stable proliferation.2,MC-LR treated liver cells, MTT was detected in cell proliferation after 24 hours.3,Analyze toxin impact upon apoptosis by FCM.4,According to results of orthogonal design, add vitamins for 24 hours in advances, then expose BRL to join MC-LR, measuring cells for ultra-oxide dismutase (SOD) and glutathione peroxidase (GSH-PX).5,RT-PCR: With the combination of orthogonal experimental design, select the effective protection portfolio by PCR technology and measure the PCNA and UDG changes at mRNA level.Results:1,Compared with normal cells, MC-LR at the concentration of less than 1μg / ml accelerate the BRL proliferation, the cell growth reached its peak at the concentration of 0.5μg / ml; MC-LR at the concentration of higher than 1.25μg / ml inhibited the cell proliferation. 2,The apoptosis changes when MC-LR's takes effect on BRL, the apoptosis rate gradually increase in the 0.001 ~ 0.5μg / ml while at 1 ~ 2.5μg / ml the rate gradually reduced.3,Compared with normal cells, under different concentrations of MC-LR, PCNA expression and UDG expression had a significant difference. At the concentration of 0.01 ~ 1.0μg / ml, PCNA expression increased in accordance with the concentration increases of MC-LR. MC-LR in the concentration of 1.25μg / ml, PCNA expression began to decline. UDG follow with PCNA expression of change.4,Orthogonal analysis showed that: compared to the cells without the protection of the vitamin, after VitB6, VitB12, VC take effects, content of SOD and GSH-PX changed, and VitB12 synergies with VC.5,When without the protection of the vitamin, cells exposed in high concentrations of MC-LR intervention, PCNA expression increased and had a significant difference after vitamin B6, B12 +VC. After vitamin B6,B12,B12 +VC role, UDG expression increased and had a significant increase.Without the protection of the vitamin cells exposed in low concentration MC-LR, PCNA expression increased and had a significant difference after vitamin B6, B12's effect; after B6, VC, B12, B12+VC effect, UDG expression decreased and had a significant difference .Conclusion:1,The effect of MC-LR in different concentrations on liver cell proliferation and on apoptosis is different, in low concentration to promote the role of cell proliferation, apoptosis rate increased gradually; in the high concentration when expressed as inhibiting cell growth or the performance of cytotoxicity , the rate of apoptosis reduced gradually.2,MC-LR changed the expression of PCNAmRNA and UDGmRNA, initiated BER repair channels, that may be the one of the repair mechanism for BRL oxidative damage.3. VitB6, VitB12, VC play a protection role in the MC-LR of BRL injury, and vitamin first order interaction for the best combination of protection: VitB12 + VC...