Effects Of Different Concentrations Of Allicin On Proliferation And Migration Of Astrocytes

Objective To investigate the effects of allicin at different concentrations on the proliferation and migration of astrocytes.Method 1.Comparison of three primary culture methods of astrocytes in rat cerebral cortex.Thirty newborn Wistar rats within 24 hours were randomly divided into three groups with 10 rats in each group.The cortical weight used in each group was almost the same.Group A:trypsinase digestion+vortex oscillation,differential centrifugation+oscillation method;Group B:trypsinase digestion,differential adherence+isothermal shaking table oscillation method;Group C:mechanical blow,differential adherence+isothermal shaking table oscillation method;Observe the cell adherence,growth status,growth rate under periodic microscope.Determination of cell purity.Counte the cells in each group after purification.The experiment was repeated three times.The experiment was repeated three times,and the culture method of group A was evaluated based on the above results.2.Effects of different concentrations of allicin on proliferation and migration of astrocytes.Astrocytes were cultured by the method of group A in part ?.Different concentrations of allicin(Control group 0 ug/ml,group A 50 ug/ml,group B 100 ug/ml,group C 200 ug/ml)were used to intervene astrocyte for 24 hours and the effects on astrocyte proliferation and migration were observed by CCK8 and scratch test.Result 1.1 The volume of cell suspensions which were extracted at the same cell density in each group:group A41.8 ± 0.15ml,group B 41.3 ± 0.1ml,group C 41.5 ± 0.1ml?Analysis of the volume of cell suspensions among the three groups:F=11.3,P=0.009 with Statistical difference.Comparisons between two groups:PA/B=0.034,PA/C=0.01 5,PB/C=0.074.The volume of cell suspensions of group A was the highest and there was significant statistical difference between group A and groupB?C.There was no statistical difference between group B and group C.1.2 observed under Microscopic:After 24 hours,group A and group B basically adhered to the bottom of culture bottles.About 30%to 40%of the cells in group C were free and a small amount of them were not adhered until the first fluid exchange.On the third day,small processes appeared in the main cells of each group,and the cell volume and processes increased with time.The typical morphological characteristics of astrocytes were observed on the seventh day.During the whole observation period,the cell morphology of group A was relatively uniform,while there were a small number of hybrid cells in group B and group C.1.3 The number of cells harvested in each group:group A(14.21 ±0.05)x107 group B(13.96±0.03)x107 group C(13.43±0.03)x107.Analyse the number of harvested cells among the three groups:F=294.118,P<0.001 with Statistical difference.Comparisons between two groups:PA/B=0.013,PA/C<0.001,PB/C=0.001.there was significant statistical difference between each two groups in the number of harvested cells.1.4 Cell purity:group A 97.87%±0.49%group B 95.90%±0.45%group C 96.38%10.59%?Analysis of purity among the three groups:F=59.258,P<0.001 with Statistical difference.Comparisons between two groups:PA/B<0.001,PA/C<0.001,PB/C=0.053?The purity of group A was the highest and there was significant statistical difference between groupA andgroupB?C.There was no statistical difference between group B and group C.2.1 The OD450 of CCK8:control group 1.12 ±0.03,groupA 0.85±0.03,groupB 0.26±0.03 group C 0.21 ±0.02.Analysis of OD450 among the four groups:F=918.4,P<0.001 with Statistical difference.Comparisons between two groups:P control/allicin all<0.001,PA/B=0.000064,Pa/c=0.000044,PB/C=0.091056.there was a statistical difference between group A and group BC,but there was no statistical difference between group B and group C.2.2 Scratch test results showed that the healing rate of 6 h,12 h and 24 h scratches was significantly different under the intervention of different concentrations of allicin,with statistical significance(P<0.001).With the increase of concentration and time,the inhibitory effect of allicin on astrocyte migration was significantly enhanced.In group D,some astrocytes were found to float and die,and gradually increased with time.Conclusion 1.Group A used Trypsinase digestion+vortex oscillation and differential centrifugation+oscillation for primary astrocyte culture.The operation was simple,the number of cells extracted was the largest,the purity was the highest,and the number of cells harvested was the largest.The method is worth promotion.2.Allicin can inhibit the proliferation and migration of astrocytes,and is positively correlated with the concentration in a certain range of concentration.200 ug/ml of allicin has shown certain cytotoxicity.