高血压通过UCP2下调介导的线粒体功能障碍加重大鼠术后学习记忆损伤

doi:10.12122/j.issn.1673-4254.2025.04.07

摘要/Abstract

摘要：

目的探索高血压与术后认知功能障碍的关系及其可能的机制。方法采用七氟烷麻醉下颈动脉暴露手术构建术后学习记忆损伤动物模型，将12周龄自发性高血压大鼠（SHR）和正常血压对照Wistar-Kyoto 大鼠（WKY）根据是否手术分为4组，即 WKY+CON组、WKY+SUR组、SHR+CON组、SHR+SUR组，8只/组。行为学测试评估大鼠术后认知变化；生物学检测炎症反应和氧化应激水平。结果高血压大鼠术后发生了明显学习记忆损伤，表现为新物体识别指数明显降低（P<0.05）；场景相关和声音相关恐惧测试僵直时间均缩短（P<0.05）。与WKY大鼠相比，SHR大鼠术后海马线粒体UCP2表达和膜电位明显降低，而ATP水平显著升高（P<0.05）；血清活性氧、IL-6、IL-1β炎症因子升高（P<0.05）；此外，SHR大鼠星形胶质细胞过度激活，神经元Nissl染色阳性细胞个数明显减少，差异均有统计学意义（P<0.05）。结论高血压可加重大鼠术后学习记忆损伤，这可能与UCP2介导的线粒体功能障碍和氧化应激损伤，进一步导致星型胶质细胞过度活化和神经元损伤有关。

关键词: 术后认知功能障碍, 自发性高血压大鼠, 解偶联蛋白2, 颈动脉暴露, 高血压, 线粒体

Abstract:

Objective To explore the correlation of hypertension with postoperative cognitive dysfunction and its possible mechanism. Methods Twelve-week-old spontaneously hypertensive rats (SHRs) and Wistar-Kyoto (WKY) rats were both randomized into control group and surgical group (n=8). In the latter group, the rats received carotid artery exposure surgery under sevoflurane anesthesia to establish models of postoperative learning and memory impairment. Postoperative cognitive function changes of the rats were evaluated using behavioral tests. The hippocampus of the rats were collected for determining ATP level and mitochondrial membrane potential (MMP) and for detecting expressions of UCP2 and astrocyte markers (GFAP and NOX4) using Western blotting and immunofluorescence staining. Serum levels of ROS, IL-6, IL-1β and TNF‑α were detected using ELISA. Nissl staining was used to examine hippocampal neuronal loss in the CA1 region. Results The SHRs exhibited exacerbated learning and memory deficits following the surgery as shown by significantly reduced performance in novel object recognition tests and context-related and tone-related fear conditioning experiments. Compared with WKY rats, the SHRs had significantly decreased mitochondrial UCP2 expression and MMP in the hippocampus, increased hippocampal ATP level, and markedly increased serum levels of ROS and inflammatory factors, showing also increased activation of hippocampal astrocytes and microglia and reduced number of neurons positive for Nissl staining. Conclusion Hypertension can exacerbate major postoperative learning and memory impairment in rats possibly as a result of UCP2-mediated mitochondrial dysfunction and oxidative stress damage, which further leads to astrocyte overactivation and neuronal damage.

Key words: postoperative cognitive dysfunction, spontaneously hypertensive rats, uncoupling protein 2, carotid artery exposure surgery, hypertension, mitochondria

刘露玉, 公茂伟, 廖国松, 赵维星, 傅强. 高血压通过UCP2下调介导的线粒体功能障碍加重大鼠术后学习记忆损伤[J]. 南方医科大学学报, 2025, 45(4): 725-735.

Luyu LIU, Maowei GONG, Guosong LIAO, Weixing ZHAO, Qiang FU. Hypertension exacerbates postoperative learning and memory impairment in rats possibly due to UCP2 downregulation-mediated mitochondrial dysfunction[J]. Journal of Southern Medical University, 2025, 45(4): 725-735.

图/表 8

图1 SHR较WKY大鼠术前血压明显高

Fig. 1 Comparison of heart rate (A), systolic blood pressure (B), mean arterial pressure (C) and diastolic pressure (D) between SHRs and WKYs before surgery. ***P<0.001, ****P<0.0001.

图2 高血压诱导大鼠旷场测试中的多动行为

Fig.2 Assessment of hypertension-induced hyperactive behaviors of rats in open field test. A: Performance of line crossing. B: Total moving distance in the test. C: Time spent in center zone. D: Time spent in the periphery zone. E: Center zone moving distance. F: Periphery zone moving distance. G: Representative motion trajectory diagram of the rats in each group. H: Representative motion trajectory heat map of each group. **P<0.01, ***P<0.001, ****P<0.0001.

图3 高血压加重大鼠术后新物体识别测试中学习记忆功能损伤

Fig.3 Novel object recognition test for assessing the effect of hypertension on spatial memory impairment of the rats after surgery. A: Novel object recognition index. B: Total exploration times in the test. C: Representative motion trajectory diagrams, where the square represent an old object and the circle represent a novel object. D: Representative motion trajectory heat map. *P<0.05.

图4 高血压加重大鼠术后恐惧记忆损伤

Fig.4 Hypertension exacerbates fear memory impairment in SHRs after surgery. A: FC context-related freezing performance (green indicates freezing, yellow indicates immobility, and red indicates mobility). B: Statistical graph of context-related percent freezing time. C: Tone-related freezing performance. D: Statistical graph of tone-related percent freezing time. E: Representative motion trajectory corresponding to context-related FC test. F: Representative motion trajectory corresponding to tone-related freezing time. **P<0.01, ***P<0.001.

图5 术后SHR大鼠海马线粒体UCP2蛋白表达明显降低

Fig.5 Quantification of mitochondrial UCP2 protein expression in rat hippocampus. A: Western blotting bands of UCP2 protein and internal control COXIV in the mitochondria. B: Statistical graph of mitochondrial UCP2 protein expression in the hippocampus. *P<0.05.

图6 高血压加重大鼠术后海马线粒体功能障碍、氧化应激和炎症反应

Fig.6 Hypertension aggravates hippocampal mitochondrial dysfunction, oxidative stress, and inflammatory response after surgery. A: Hippocampal ATP level. B: Hippocampal mitochondria membrane potential (MMP). C: Reactive oxygen species level in serum. D: Serum IL-6 level. E: Serum IL-1β level. F: Serum TNF-α level. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

图7 高血压加重大鼠术后海马星型胶质细胞的过度激活

Fig. 7 Immunofluorescence staining of the hippocampus of SHR and WKY rats for detecting expressions of astrocyte markers in the CA1 region. A: Combo graph of immunofluorescence (Scale bar=20 μm; green represents GFAP, and red represents NOX4). B, C: Statistical graphs of NOX4 and GFAP. *P<0.05.

图8 高血压加重大鼠术后神经元损伤

Fig.8 Nissl staining images of different regions in rat hippocampus (A) and comparison of positive cell numbers in the CA1 region among the groups (B). **P<0.01, ***P<0.001

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