The Effects Of Keto-valine-calcium, Keto-isoleucine-calcium And Hydroxylmethionine-calcium On Human Mesangial Cell (HMC)

Object: To study the effects of keto-valine-calcium, keto-isoleucine-calcium and hydroxylmethionine- calcium on human mesangial cell(HMC)Method: HMCs were stimulated respectively by keto- valine-calcium, keto- isoleucine- calcium and hydroxylmethionine- calcium,MTT method was used to measure of HMCs' proliferation, the expressionof LDH was detected by biochemistry, the AT1R was accessed byimmunocytochemistry, TGF-β1 was observed by ELISA, and the effecton cell cycle and apoptosis was studied by flow cytomentry.Results: After stimulating by 10mM keto-valine-calcium, keto-isoleucine-calcium and hydroxylmethionine-calcium, the cells' prolix-feration were inhibited, and the expression level of LDH on the cellcultural supematant didn't increase. There were no statistically significantdifferences between DMSO control group and other study groups (p＞0.05) except for the groups of.keto-isoleucine-calcium and co-intervention (p＜0.05).After the medicine intervention, the expressionlevel of AT1R increased compared to negative and DMSO control group,however, the level significantly increased after the co-intervention anddid not increase after amphi-combination intervention compared to single medicine intervention. The expression of TGF-β1 was higher in everygroup than in DMSO control group (p＜0.01), especially in theamphi-combination groups and significally in the co-intervention group.The result of cell flow cytomentry shown that different medicineintervention groups can induce cell cycle G1 arrest and cell apoptosis. Toour surprise, there was no statistically different in the single medicineintervention, while the apoptosis ratio was increased in the amphi-combination groups especially in the co-intervention group (18.76±1.02)compared to DMSO control group (1.09±0.33),visibly.Conclusion: 10mM keto-valine-calcium, keto-isoleucine- calciumand hydroxylmethionine-calcium have multiple effects on humanmesangial cell (HMC). They not only increase the expression of AT1R,but also add the expression of TGF-β1. Furthermore, they can induce cellcycle G1 arrest and cell apoptosis.