The Effect Of Free Gossypol On Mus Uropoietic System

Four experiments were conducted to study the effect of free gossypol on Mus uropoietic system.Experiment 1.The effect of different concentration gradient free gossypol on Mus body weight and uropoietic system. select body weight 24.69±1.01g Kunming Mus musculus albus 90 and divide into 5 groups randomly. 0 mg/kgBW*d, 40 mg/kgBW*d, 80 mg/kgBW*d, 120 mg/kgBW*d,160 mg/kgBW*d. 18 Muses every group, and divide into 3 boxes, 6 Muses per group. Pretesting 7 days, trialing 6 weeks. Observe the Mus healthy state, to measure the body weight of every group in every weeks, collection the kidney and do HE microtome section which observed by microscope. The results indicate that, the effect of gossypol on Mus'growth is not manifest in the first 3 weeks, but it can inhibit the growth speed evidently in anaphase. Gossypol can induce Mus Renal interstitium hemorrhage, renal tubular epithelial cell Granular degeneration cellular, Infarction of kidney, Glomerular capillary hemorrhage, bloodvessel dilatationa and so on.Experiment 2. The effect of gossypol on Mus'K, Mg, Ca, P in blood serum. At 2 weeks and 6 weeks of experiment, collecte the blood of every group ,and test the dosage of K, Mg, Ca and P in blood serum. The results indicate that, feed mus with 40-160 mg/kgBW*d concentration free gossypol can induce hypopotassemia, but the Mg, Ca and P are not obviously.Experiment 3. Isolated and culture KM Mus musculus albus primary renal tubular epithelial cell, And through cell mediated immunity to identify the renal tubular epithelial cell .The results show that , The yield of primary tubular epithelial cells by adult group were better than suckling group;the two enzyme digestion methods were suitable in 10 min'digestion. The collagenase digestion method is better than trypsinase digestion method in cells adherence rate and quantity; identity result is positive.Experiment 4. The effect of gossypol on renal tubular epithelial cell of Mus. Add 31.25~1000μg/ml (±)gossypol and (-)gossypol to cells expectively, when the cells get in touching with gossypol 24h,48h,72h and 96h, use MTT test apoptosis, with kit respectively test HDL and ultramicro-ATPase including CMA and NKA. The results show that, the apoptosis gets higher with concentration of (±)gossypol adding more and the touching time longer. But the (-)gossypol except 24h action time incommon with (±)gossypol, in other action times low concentration (-)gossypol compare to the high concentration is not distinguished. During 24 h, the LDH divulge dosage of (±)gossypol is higer than (-)gossypol, and the LDH divulge dosage get higher according to the (±)gossypol concentration increace. But the NKA activity get stronger with the (-)gossypol concentration increace, and at the same action time , the effect of (-)gossypol on NKA activity is stronger than (±)gossypol. The effect of high dose gossypol is significant on CMA activity. At 24 h ,the CMA vigor get lower with the (-)gossypol concentration increace. So that the effet of (-)gossypol is stronger compare to (±)gossypol on renal tubular epithelial cells. But in 31.25-1000μg/ml concentration, there is no obvious relying relationship between gossypol concentration and enzyme activity.Conclusion:Mus musculus albus feeding with free gossypol, can inhibit the increasing of body weight, can leed to the damage of Mus kidney tissue, and result in hypopotassemia of Mus.When gossypol touching with gossypol, the (±)gossypol and (-)gossypol can leed to renal tubular epithelial cell apoptosis and damage respectively, induce the HDL enzymatic divulging, lead to the NKA and CMA enzyme activity depressing. And the (-)gossypol can effect more severe than (±)gossypol on NKA enzyme activity.