| Purpose To observe real-time changes of intracellular calcium arose from bilirubinand GUDCAin synaptosome from brainstem in the SD rat.Methods Fourty P7-14SD rats (male and female)were used in this study.They wererandomly assigned to two groups. A group was bilirubin group and B group was bulirubinplus GUDCA group. Twenty rats were in each group,five rats were in every differentconcentration. Rats were decapitated after anesthetized with intramuscular injection ofacepromazine(4.5mg/kg of body weight).Then, the brainstem were dissected.Thesynaptosomes were purified by sucrose density gradient centrifugation. After loadingOG-BAPTAin synaptosomes from brainstem（45min,room temperature）, two dimensionalimage of intracellular calcium and analysis of fluorescence intensity were achieved byConfocal laser scanning microscopy(CLSM)．Results Compared with blank group, an immediate peak of fluorescent intensityof [Ca2+]i was detected as soon as bilirubin was applied,and increased depended onthe concertration. Fluorescent intensity of[Ca2+]i was reduced obviously using GUDCA.Conclusions The results indicated that GUDCA can inhibit bilirubin-induced calciumoverload in synaptosomes. GUDCA might be a valuable preventive therapy in neonates atrisk of UCB encephalopathy. |
