The Effects Of Constant High-glucose、Fluctuating Glucose And Exenatide On The Apoptosis Of Human Renal Mesangial Cells Mediated By JNK Signaling Pathway

Objective:1、To culture human renal mesangial cells in vitro and to observe theirs growth status2、To observe the effects of normal glucose,persistent high glucose and fluctuating glucose on human renal mesangial cells growth.To compare the differences in the apoptosis rate and the proliferation rate.3、To explore the mechanism of JNK signaling pathway in the apoptosis of human renal mesangial cells and the anti-apoptotic effect of the glp-1 receptor agonist.4、To explore the protective effect of the glp-1 receptor agonist on the human renal mesangial cells in vitro,and explore its mechanism to provide theoretical basis for the clinical treatment of diabetic nephropathy.Methods:1、we cultured human renal mesangial cells in vitro and observed cell morphology by inverted microscope.2、Human renal mesangial cells can be randomly divided into 6 groups.group 1: with normal glucose(5.6mmol/ L)medium culture,group 2: with persistent high glucose(25mmol/L)medium culture,group3: with fluctuating glucose(5.6 mmol/L,25mmol/L)medium culture(first culture the cells with high glucose medium culture for 3 h,then with normal glucose medium culture for 2 h,repeat 3 times in a day,and at last overnight in normal glucose culture medium),group4: with normal glucose(5.6mmol/ L)medium culture based on the final concentration of 100 nmol/L of exenatide,group5: with persistent high glucose(25mmol/L)medium culture based on the final concentration of 100 nmol/L of exenatide,group6:with fluctuating glucose(5.6 mmol/L,25mmol/L)medium culture based on the final concentration of100 nmol/L of exenatide.The six groups were all intervened for 24 hours respectively.The morphological changes were observed by the inverted microscope,and the photos were recorded.cck8 method was used to determine the rates of the proliferation.Rt-pcr method was used to determine the relative expression of c-junmRNA in each groups.Western blot method was used to determine the relative expression of CJUN protein in each groups.SPSS software was used for statistical.Results:1.Human renal mesangial cells cultured in vitro were in good condition.2.Observation under inverted microscope: No obvious difference was observed under low power microscope.The cell morphology was slightly irregular and the cell membrane was slightly blurred in group5 and group6,also the intracellular granules were increased.3 cck8: The proliferation rate of group1 is greater than group3 and group5,and the differences were statistically significant.The proliferation rate of group3 is greater than group5,but there was no statistically significant difference.There were no statistically significant differences between drug additive groups and non-additive groups.4,RT-PCR::There were no statistically significant differences between the three groups.Also we didn’t get statistically significant differences between drug additive groups and non-additive groups5,Western blot:There were no statistically significant differences between the three groups.Also we didn’t get statistically significant differences between drug additive groups and non-additive groups6.Conclusion: Under the stimulation of persistent high glucose and fluctuating glucose,we are not sure wheather the endoplasmic reticulum stress associated apoptotic pathway in human renal mesangial cells was activated in different degrees.