《Table 1 Details of the primers used for RT-PCR》

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《Inhibition of α5 GABAA receptors has preventive but not therapeutic effects on isoflurane-induced memory impairment in aged rats》

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RT-PCR:Reverse transcription-polymerase chain reaction；α5GABAAR:α5 subunit-containing gamma-amino butyric acid type Areceptors.

RT-PCR was performed to investigate the influence of isoflurane and L-655，708 on the mRNA expression ofα5GABAARs in the hippocampus.Following behavioral tests，rats were sacrificed，and hippocampal tissue was obtained and stored at–80°C immediately.Then the lysed tissue was homogenized by a homogenizer（Qiagen，Inc.，Valencia，CA，USA）and then centrifuged.Total RNA was acquired from the supernatant and its concentration was measured using a Nano Drop ND-1000 Spectrophotometer（Thermo Scientific，Wilmington，DE，USA）.α5 GABAAR andβ-actin primers were purchased from Tianyi Biotech，Beijing，China and sequence information was shown in Table 1.RT-PCRwas conducted using the real-time PCR system with SYBR?Premix（TaKaRa，Inc.，Dalian，China）.α5 GABAAR mRNAlevels were standardized toβ-actin levels as internal controls.PCR was conducted by 40 cycles at 96°C for 30 seconds，55°Cfor 90 seconds，and 72°C for 60 seconds in a BioRad CFX96?Real-Time System C1000TM Thermal Cycler（BioRad Laboratories，Shanghai，China）.Threshold cycle（Ct）values were recorded and tested.The Ct values were defined as the amplification cycles when the fluorescent signal in the reaction system significantly exceeded the background fluorescence（threshold）.The Ct values were negatively related to the tested mRNA level.Consequently，a lower Ct reflects a higher mRNA level（Dhar et al.，2002；Park et al.，2015）.