Different Concentrations Of Uric Acid Salt Of 3 T3 L1 And Ins - 1 Cell Inflammation And Oxidative Stress

Objective:To observe the role of different concentration of urate in the expression of adipose cell injury associated factors; To identify The mechanism of adipose cell damage by urate.Method:The international method was used on3T3-L1adipocytes differentiating to mature adipocytes and the oil red O staining was used to identify the cell differentiation. After cell count, the cells were individed into ten groups as normal group, low concentration groups(5mg/dL、7mg/dL、9mg/dL),medium concentration groups(12mg/dL、15mg/dL、18mg/dL) and high concentration groups(21mg/dL24mg/dL,27mg/dL). All of the groups stimulated adipocytes for48hours.levels of IL-1、 TNF-a、 AGEs in the supernatants of cell cultures were measured by enzyme linked immunosorbent assay(ELISA) and residual cells did MTT assay.Results①When the urate concentration were in5mg/dL and7mg/dL, the level of SOD was markly increased than of control group.When15mg/dL and more, SOD were significantly decreased, however MDA increased(P<0.05).②When the urate concentration were in9mg/dL to18mg/dL, the level of IL-1βwere markly decreased than of the control group(P<0.05). The affection of urate on adipocyte TNF-a was not obvious, only when the urate concentration were21mg/dL resulted in adipocyte viability decreased, the TNF-alevel decreased significantly(P<0.05).③When the urate concentration were21mg/dL, the absorbance of cells significantly decreased,and27mg/dL, the absorbance were the lowest(P<0.05).Conclusions:In vitro, when the urate concentration were in5mg/dL and7mg/dL, urate has a certain role by increasing level of SOD. However, when urate concentration gets up to15mg/dL, adipocytes injuried by SOD decreased and MDA increased. Then the process is exactly along with the inflammatory response, further research is needed to be confirmed. Objective:To observe the role of different concentration of urate in the expression of islet cells injury associated factors; To identify The mechanism of islet cells damage by urate.Method:After culturing cell, the cells were individed into six groups as normal group, urate low concentration groups(5mg/dL、7mg/dL),medium concentration groups(9mg/dL) and high concentration groups(12mg/dL、15mg/dL). All of the groups stimulated urate for24hours.After getting supeunatant, cells were incubated with KRHB buffer with different concentrations of glucose, levels of IL-1β、 TNF-α、SOD、 MDA and insulin in the supernatants of cell cultures were measured by enzyme linked immunosorbent assay(ELISA) and residual cells did MTT assay.Results①When the urate concentration were5mg/dL, SOD was significantly increased than of the control group (P<0.05);When the urate concentration were9mg/dL and12mg/dL, MDA were significantly increased than of the control group (P<0.05), however SOD no significantly decreased, When the urate concentration were9mg/dL to15mg/dL(P<0.05);②When the urate concentration were9mg/dL and12mg/dL, IL-1βand TNF-a were markly increased than of the control group (P<0.05);③When the urate concentration were12mg/dL, the absorbance of cells significantly decreased (P<0.05).④When the urate concentration were12mg/dL,the basic secretory function of islet cels was decreased than the control group;When7mg/dL, the secretory function after stimulating by the high glucose was significantly decreased.Conclusions:In vitro, when the urate concentration were in5mg/dL, urate has a certain role of resisting oxidative stress by increasing level of SOD. However, when urate concentration gets up to9mg/dL, islet cells injuried by inflammation and oxidative stress, which reduce the sensitivity of the islet cells to glucose.