| 江 山,徐成峰,李娅娜,谢红萍,袁 华,王会会,李 玲.次声刺激诱导原代培养大鼠星形胶质细胞释放谷氨酸的研究[J].中国康复医学杂志,2014,29(6):499~503 |
| 次声刺激诱导原代培养大鼠星形胶质细胞释放谷氨酸的研究 点此下载全文 |
| 江 山 徐成峰 李娅娜 谢红萍 袁 华 王会会 李 玲 |
| 解放军总医院第一附属医院，北京海淀区阜成路51号，100048 |
| 基金项目：国家自然科学基金资助项目（81071595, 81201514） |
| DOI： |
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| 全文下载次数: 2030 |
| 摘要： |
| 摘要
目的：观察16Hz, 130dB次声刺激对原代培养的大鼠星形胶质细胞谷氨酸释放的影响，并探讨其释放机制。
方法：原代培养大鼠海马区星形胶质细胞，将其分为次声刺激组（IE group，n=6），Gap26+次声刺激组（Gap26+IE group），对照组（Ctrl group，n=6）。次声刺激组细胞暴露于次声舱中，分别给予15min, 30min, 60min, 90min, 120min和240min的次声刺激；对于Gap26+IE组，则在次声刺激前，选用Cx43半通道阻断剂Gap26预处理星形胶质细胞；对照组也置于次声舱，但不给予次声刺激。次声刺激频率为16Hz，强度为130dB。为了排除Gap26对谷氨酸释放的影响，还选用乱序Gap26肽段(Scrb Gap26)来预处理星形胶质细胞（Scrb Gap26+IE group，n=6）。采用免疫荧光染色测定细胞Cx43的表达情况，采用高效液相色谱（HPLC）来测定细胞外液谷氨酸浓度。
结果：次声刺激后，IE组细胞外液谷氨酸的浓度显著升高，并在刺激90min后，细胞外液谷氨酸浓度达峰值，为（4.6±0.3）nmol/ml，显著高于对照组（2.3±0.2）nmol/ml（P<0.05），这说明次声刺激可以显著增高星形胶质细胞外液的谷氨酸含量。此外，次声刺激后，IE组细胞Cx43的表达也显著升高，刺激60mins时，Cx43平均荧光强度达到峰值，为（198±33）(AUC)，显著高于对照组（P<0.05）。而对于Gap26+IE组，次声刺激后细胞外液谷氨酸浓度的升高受到抑制，90min后，细胞外液谷氨酸浓度为（3.58±0.17）nmol/ml，显著低于次声刺激组（P<0.05）。
结论：次声刺激可以诱导星形胶质细胞释放谷氨酸；Cx43半通道阻断剂Gap26抑制了谷氨酸释放，次声刺激诱导的谷氨酸释放可能是通过Cx43半通道来完成的。 |
| 关键词：次声 星形胶质细胞 谷氨酸 Cx43半通道 |
| Infrasound induced glutamate release by cultured astrocytes of rats Download Fulltext |
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| First Affiliated Hospital, Chinese PLA General Hospital, Beijing,100048 |
| Fund Project: |
| Abstract: |
| Abstract
Objective:To detect the effect of infrasound on the glutamate release by cultured astrocytes of rats and the possible release routes.
Method:Cultured hippocampal astrocytes were divided into three groups: control group(Ctrl group), infrasound exposure group(IE group) and Gap26+ IE group. The astrocytes in the IE group were placed into the infrasonic chamber and exposed to infrasound(16Hz, 130dB) for different durations(15min, 30min, 60min, 90min, 120min, 240min), while Ctrl group was put into the same infrasonic chamber but without infrasonic exposure. Furthermore, in order to identify the glutamate release route under infrasound, astrocytes were pre-treated with Cx43 hemichannel blocker Gap26 before infrasound exposure. In addition, scrambled Gap26 peptide(Scrb Gap26) was used to rule out the effect of the Gap26 itself on the glutamate, which was regarded as the Scrb Gap26+IE group. After infrasound exposure, immunocytochemistry method was used to detect the expression of Cx43. At the same time, high pressure liquid chromatography(HPLC) was used to detect the extracellullar glutamate level.
Result：In IE group, the extracellular glutamate level increased after infrasound exposure. At 90min after IE, the extracellular glutamate level increased to the maximum of （4.6±0.3）nmol/ml（n=6）, which was significantly higher than that of Ctrl group(2.3±0.2）nmol/ml(P<0.05 vs Ctrl group). The infrasound exposure increased the expression of astroglial Cx43 significantly. At 60min after IE the mean fluorescence intensity(MFI) of Cx43 was about （198±33）AUC（area under the curve）, which was higher than that of ctrl group（P<0.05 vs Ctrl group）. The increase can be inhibited by the pretreatment of Gap26. In Gap26+IE group, the extracellular glutamate level was （3.58±0.17）nmol/ml at 90 min after IE, which was significantly lower than that of IE group（P<0.05 vs IE group）.
Conclusion: Glutamate release by Cx43 hemichannels is involved in the response of cultured astrocytes to infrasound. Cx43 hemichannel blocker Gap 26 can inhibit the infrasound induced glutamate release. |
| Keywords:infrasound astrocyte glutamate Cx43 hemichannel |
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