Studies On Apoptosis Of Sow Luteal Cells Induced By Gossypol In Primary Culture

China is a main country that produces a large quantity of cotton. There is a wide source of cottonseed cake with a high nutrition value and low price. Cottonseed cake has become an important source of plant protein in feedstuff gradually. Gossypol is a main poisonous ingredient existing in cottonseed cake, which has been testified by previous studies as a kind of anti-fertility substance for male animals, but there is a little study on reproductive toxicity of gossypol to the female animals with different conclusion. In order to utilize cottonseed cake as feedstuff rationally, we performed this research.Sow primary luteal cell is used as empirical model. Through MTT assay, agarose gel electrophoresis of DNA and TUNEL assay, we studied on apoptosis and toxicity on sow luteal cell treated with 0, 0.4, 2, 10, 50μg/mL gossypol. The luteal cells, separated by a synthetical application of a mechanical separating way and an assimilation of 0.2% Collagenase II, the survival rate of which is above 90% testified by 0.4% trypan blue dye test, can meet the empirical demand after incubating 24 h. Testify the restraining effect of gossypol to the reproduction of sow primary luteal cell by MTT assay, the results show: after 24 h and 48 h, the group treated by 5% alcohol shows no difference (P>0.05) contrasted with the group treated by Oug/mL gossypol, which indicated the effect of 5% alcohol to the reproduction of sow primary luteal cells can be ignored. After 24 h, the group treated with 0.4μg/mL gossypol has a remarkable difference (P<0.05) contrasted with the group of 5% alcohol, other groups of different concentration of gossypol have remarkable difference significantly contrasted with the control group (P<0.01). After 48 h, the all groups of different concentration of gossypol have remarkable difference significantly contrasted with the control group. Contrasted with the same concentration of gossypol in two periods, the groups of 10μg/mL and 50μg/mL have remarkable difference significantly. After 48 h, the restraining rates of the two kind of concentration of gossypol to sow primary luteal cells arrive 60% and 70% respectively. DNA Ladder can't be found in the agarose gel electrophoresis. The test of TUNEL assay show: there are apoptosis of sow luteal cells treated by various gossypol, the rates of apoptosis are 5%-23% and increase with the increase of concentration of gossypol. The all groups of different concentration of gossypol after 24 h and 48 h have remarkable difference significantly contrasted with the control group. The two groups of 0.4μg/mL and 2μg/mL in same...