Background Isoflurane, a commonly used inhaled anesthetic, induces apoptosis in primary rat cortical neurons of rat in a concentration- and time-dependent manner by an unknown mechanism. We hypothesized that isofluran...Background Isoflurane, a commonly used inhaled anesthetic, induces apoptosis in primary rat cortical neurons of rat in a concentration- and time-dependent manner by an unknown mechanism. We hypothesized that isoflurane induced apoptosis by causing abnormal calcium release from the endoplasmic reticulum （ER） via activation of inositol 1,4,5-trisphosphate （IP3） receptors. Sevoflurane has a reduced ability to disrupt intracellular calcium homeostasis and is a less potent cytotoxic agent. This study examined and compared the cytotoxic effects of isoflurane and sevoflurane on rat primary cortical neurons and their relationship with disruption of intracellular calcium homeostasis and production of reactive oxygen species （ROS）.Methods Primary rat cortical neurons were treated with the equivalent of 1 minimal alveolar concentration （MAC） of isoflurane and sevoflurane for 12 hours. MTT reduction and LDH release assays were performed to evaluate cell viability. Changes of calcium concentration in the cytosolic space, [Ca^2＋]c, and production of ROS were determined after exposing primary rat cortical neurons to isoflurane and sevoflurane. We also determined the effects of IP3 receptor antagonist xestospongin C on isoflurane-induced cytotoxicity and calcium release from the ER in primary rat cortical neurons.Results -Isoflurane at 1 MAC for 12 hours induced cytotoxicity in primary rat corticai neurons, which was also associated with a high and fast elevation of peak [Ca^2＋]c. Xestospongin C significantly ameliorated isoflurane cytotoxicity in primary cortical neurons, as well as inhibited the calcium release from the ER in primary cortical neurons. Isoflurane did not induce significant changes of ROS production in primary rat cortical neurons. Sevoflurane, at equivalent exposure to isoflurane, did not induce similar cytotoxicity or elevation of peak [Ca^2＋]c in primary rat cortical neurons.Conclusion These results suggested that isoflurane induced elevation in [Ca^2＋]c, partially via elevated activit展开更多
目的建立人网膜前脂肪细胞原代培养的方法,研究内脏脂肪增生肥大和内分泌功能的生物学特性。方法选取人网膜脂肪组织,采用酶消化细胞原代培养出梭形细胞,对培养细胞进行形态学观察、MTT 法测定生长曲线、油红 O 脂肪染色法进行脂肪细胞...目的建立人网膜前脂肪细胞原代培养的方法,研究内脏脂肪增生肥大和内分泌功能的生物学特性。方法选取人网膜脂肪组织,采用酶消化细胞原代培养出梭形细胞,对培养细胞进行形态学观察、MTT 法测定生长曲线、油红 O 脂肪染色法进行脂肪细胞鉴定、酶联免疫法测定脂肪细胞因子瘦素和脂联素水平。结果培养出的梭形细胞成分均一。第3天开始增殖,4～9 d 为指数增长期,倍增时间约为60 h。经分化诱导21d 约有90%的细胞分化为成熟脂肪细胞。前脂肪细胞可检出低水平瘦素分泌,经诱导分化分泌量持续增多,第17天达峰值(1.6 ng·ml^(-1)·24 h^(-1)),之后保持高分泌状态至诱导结束;而脂联素直到诱导分化第7天始可检测到低水平分泌,此时光镜下已可见胞质含脂滴的细胞;7～17d 分泌逐渐增多,第17天分泌达峰值(99 ng·ml～·24 h^(-1)),之后有明显下降趋势。经油红 O 脂肪染色和瘦素、脂联素分泌检测证实分离培养的细胞为功能活跃的前脂肪细胞。结论成功建立了人网膜前脂肪细胞模型,观察到瘦素和脂联素不同分泌模式。脂联素可作为鉴定前脂肪细胞分化成熟的内分泌功能特异标志物。展开更多
文摘Background Isoflurane, a commonly used inhaled anesthetic, induces apoptosis in primary rat cortical neurons of rat in a concentration- and time-dependent manner by an unknown mechanism. We hypothesized that isoflurane induced apoptosis by causing abnormal calcium release from the endoplasmic reticulum （ER） via activation of inositol 1,4,5-trisphosphate （IP3） receptors. Sevoflurane has a reduced ability to disrupt intracellular calcium homeostasis and is a less potent cytotoxic agent. This study examined and compared the cytotoxic effects of isoflurane and sevoflurane on rat primary cortical neurons and their relationship with disruption of intracellular calcium homeostasis and production of reactive oxygen species （ROS）.Methods Primary rat cortical neurons were treated with the equivalent of 1 minimal alveolar concentration （MAC） of isoflurane and sevoflurane for 12 hours. MTT reduction and LDH release assays were performed to evaluate cell viability. Changes of calcium concentration in the cytosolic space, [Ca^2＋]c, and production of ROS were determined after exposing primary rat cortical neurons to isoflurane and sevoflurane. We also determined the effects of IP3 receptor antagonist xestospongin C on isoflurane-induced cytotoxicity and calcium release from the ER in primary rat cortical neurons.Results -Isoflurane at 1 MAC for 12 hours induced cytotoxicity in primary rat corticai neurons, which was also associated with a high and fast elevation of peak [Ca^2＋]c. Xestospongin C significantly ameliorated isoflurane cytotoxicity in primary cortical neurons, as well as inhibited the calcium release from the ER in primary cortical neurons. Isoflurane did not induce significant changes of ROS production in primary rat cortical neurons. Sevoflurane, at equivalent exposure to isoflurane, did not induce similar cytotoxicity or elevation of peak [Ca^2＋]c in primary rat cortical neurons.Conclusion These results suggested that isoflurane induced elevation in [Ca^2＋]c, partially via elevated activit
文摘目的建立人网膜前脂肪细胞原代培养的方法,研究内脏脂肪增生肥大和内分泌功能的生物学特性。方法选取人网膜脂肪组织,采用酶消化细胞原代培养出梭形细胞,对培养细胞进行形态学观察、MTT 法测定生长曲线、油红 O 脂肪染色法进行脂肪细胞鉴定、酶联免疫法测定脂肪细胞因子瘦素和脂联素水平。结果培养出的梭形细胞成分均一。第3天开始增殖,4～9 d 为指数增长期,倍增时间约为60 h。经分化诱导21d 约有90%的细胞分化为成熟脂肪细胞。前脂肪细胞可检出低水平瘦素分泌,经诱导分化分泌量持续增多,第17天达峰值(1.6 ng·ml^(-1)·24 h^(-1)),之后保持高分泌状态至诱导结束;而脂联素直到诱导分化第7天始可检测到低水平分泌,此时光镜下已可见胞质含脂滴的细胞;7～17d 分泌逐渐增多,第17天分泌达峰值(99 ng·ml～·24 h^(-1)),之后有明显下降趋势。经油红 O 脂肪染色和瘦素、脂联素分泌检测证实分离培养的细胞为功能活跃的前脂肪细胞。结论成功建立了人网膜前脂肪细胞模型,观察到瘦素和脂联素不同分泌模式。脂联素可作为鉴定前脂肪细胞分化成熟的内分泌功能特异标志物。
