The Effect Of High Glucose Environment On Iccs' Morphology And Ultrastructural In Guinea Pig Bladder Cells

Purpose:To explore the damage caused by high glucose environment in the way of observing guinea pig bladder ICCs'morphologic and ultra structure changes in high glucose environment.Methods:With collagenase digestion in vitro, we get the primary culture ICCs body outside. The ultra structure of ICCs was observe by transmission electron microscope in different glucose concentrations (5,10,15mmol/L) and different times(24,72h), while take Fluo-4AM loaded ICCs, record their length and the variety of calcium ion by confocal laser scanning technology.Results:With increased glucose concentration and prolonged culture time, the amount of organelles in ICCs decreased, which embodied in changes of mitochondrion and endocytoplasmic reticulum under transmission electron microscope. The mitochondrion was different in size, hydropic changed, swelling, and even lysed. The endocytoplasmic reticulum was expanded, hydropic changed, the cytoplasm widely lysed and the enation disappeared. Bridge-like structure can also be observed in part of ICC. With high glucose concentration and long culture time, the ICCs can be divided into two types by staining. One type was hypochromatic and filarin was found in cells, the other type was trachychromatic and liquefactive necrosis was found in cells. Under confocal microscope, with increased glucose concentration and prolonged culture time, fluorescence strength of calcium ion in the primary cultured ICCs was as follows: calcium ion of 5mmol/L/24h group has no significant difference when compared to that of 5mmol/L/72h group; it was increased in 10mmol/L/24h group and decreased in 15mmol/L/24h group when compared to that of 5mmo1/L/24h group; calcium ion in both 10mmol/L72h group and 15mmol/L/72h group was increased, however,5mmol/L/72h group was lower than that of 10mmol/L/72h,which begin to show the trend of decrease. The length of ICCs in 5mmol/L/24h group and 5mmol/L/72h group have no significant difference; in 24h groups, the length of ICCs in 10mmol/L group and 15mmol/L group were shortened as glucose concentration; in 10mmol/L/72h groups the length of ICCs was increased while in 15mmol/L/72h groups was decreased.Conclusion:The high glucose environment, which can damage ICCs'morphologic and ultra structure changes, leads to changes of calcium ion inflow, which caused the pacing decrease or vanish. This changes of ICCs would be one of the important pathologic bases of DCP.