Department of Anesthesiology, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, No. 107 Yanjiang West Road, Guangzhou, 510120, China.
Guangdong Province Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, 510120, China.
Neurochem Res. 2021 Mar;46(3):468-481. doi: 10.1007/s11064-020-03180-2. Epub 2020 Nov 25.
General anesthetics interfere with dendritic development and synaptogenesis, resulting in cognitive impairment in the developing animals. RhoA signal pathway plays important roles in dendritic development by regulating cytoskeleton protein such as tubulin and actin. However, it's not clear whether RhoA pathway is involved in inhaled general anesthetics sevoflurane-induced synaptic development abnormalities and long-term cognitive dysfunction. Rats at postnatal day 7 (PND7) were injected intraperitoneally with RhoA pathway inhibitor Y27632 or saline 20 min before exposed to 2.8% sevoflurane for 4 h. The apoptosis-related proteins and RhoA/CRMP2 pathway proteins in the hippocampus were measured 6 h after sevoflurane exposure. Cognitive functions were evaluated by the open field test on PND25 rats and contextual fear conditioning test on PND32-33 rats. The dendritic morphology and density of dendritic spines in the pyramidal neurons of hippocampus were determined by Golgi staining and the synaptic plasticity-related proteins were also measured on PND33 rats. Long term potentiation (LTP) from hippocampal slices was recorded on PND34-37 rats. Sevoflurane induced caspase-3 activation, decreased the ratio of Bcl-2/Bax and increased TUNEL-positive neurons in hippocampus of PND7 rats, which were attenuated by inhibition of RhoA. However, sevoflurane had no significant effects on activity of RhoA/CRMP2 pathway. Sevoflurane disturbed dendritic morphogenesis, reduced the number of dendritic spines, decreased proteins expression of PSD-95, drebrin and synaptophysin, inhibited LTP in hippocampal slices and impaired memory ability in the adolescent rats, while inhibition of RhoA activity did not rescue the changes above induced by sevoflurane. RhoA signal pathway did not participate in sevoflurane-induced dendritic and synaptic development abnormalities and cognitive dysfunction in developing rats.
全身麻醉会干扰树突的发育和突触形成,导致发育期动物的认知障碍。RhoA 信号通路通过调节微管蛋白和肌动蛋白等细胞骨架蛋白在树突发育中发挥重要作用。然而,RhoA 通路是否参与吸入性全身麻醉药七氟醚诱导的突触发育异常和长期认知功能障碍尚不清楚。在暴露于 2.8%七氟醚 4 小时之前,将出生后第 7 天(PND7)的大鼠腹膜内注射 RhoA 通路抑制剂 Y27632 或生理盐水 20 分钟。在七氟醚暴露后 6 小时测量海马中的凋亡相关蛋白和 RhoA/CRMP2 通路蛋白。在 PND25 大鼠上进行旷场试验和 PND32-33 大鼠上进行情境恐惧条件试验来评估认知功能。通过 Golgi 染色测定海马锥体神经元的树突形态和树突棘密度,并在 PND33 大鼠上测量突触可塑性相关蛋白。在 PND34-37 大鼠上记录海马切片的长时程增强(LTP)。七氟醚诱导 PND7 大鼠海马 caspase-3 激活,降低 Bcl-2/Bax 比值并增加 TUNEL 阳性神经元,而 RhoA 抑制可减轻这些变化。然而,七氟醚对 RhoA/CRMP2 通路的活性没有显著影响。七氟醚扰乱树突形态发生,减少树突棘数量,降低 PSD-95、drebrin 和突触素的蛋白表达,抑制海马切片中的 LTP,并损害青少年大鼠的记忆能力,而 RhoA 活性抑制不能挽救七氟醚诱导的上述变化。RhoA 信号通路不参与发育期大鼠七氟醚诱导的树突和突触发育异常及认知功能障碍。