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目的 基于PI3K-Akt通路探讨补阳还五汤（BHD）对急性高原缺氧性脑损伤模型小鼠的保护作用及抗凋亡机制。方法 将90只健康雄性C57BL/6J小鼠随机分为空白对照组（NC组）、低氧模型组（M组）、补阳还五汤低、中、高剂量组（BHD-L/M/H组）及地塞米松组（Dex组），除NC组外，其余各组分别给予相应药物灌胃。灌胃第5天，除NC组外，将其余小鼠置于模拟6 000 m海拔的低压氧舱中持续72 h以建立急性高原缺氧性脑损伤模型。通过IntelliCage全自动智能监测系统评估小鼠认知功能；采用HE染色和透射电子显微镜观察小鼠脑组织形态学与线粒体超微结构；通过尼氏染色观察海马神经元数目，通过TUNEL染色检测小鼠神经元凋亡情况；采用ELISA检测相关血清标志物（S100B蛋白、神经胶质细胞原纤维酸性蛋白）水平；通过RT-qPCR检测磷脂酰肌醇3激酶（PI3K）、蛋白激酶B(Akt)、Bcl-2、Bax及胱天蛋白酶3的基因转录水平；通过Western blot检测磷酸化PI3K(p-PI3K)、磷酸化Akt(p-Akt)、Bcl-2、Bax及活化型胱天蛋白酶3的蛋白水平。结果 在行为学测试鼻触学习阶段、空间逆转阶段及新奇事物阶段，与NC组相比，M组小鼠鼻触次数与访问次数均减少（均P<0.01），出现认知功能下降；形态学结果显示海马神经元结构被破坏、线粒体嵴断裂、细胞凋亡率显著升高（P<0.001），同时伴随血清S100B蛋白与神经胶质细胞原纤维酸性蛋白水平升高（均P<0.01）；在分子层面，M组p-PI3K、p-Akt及Bcl-2的蛋白及mRNA水平下调，而Bax与活化型胱天蛋白酶3的表达上调（均P<0.05）。与M组相比，BHD-H组和Dex组访问与鼻触次数增加（均P<0.05）。BHD各剂量组的指标均得到恢复，其中BHD-H组的效果最为显著（P<0.05），且与Dex组效果相当（P>0.05）。结论 补阳还五汤可改善急性高原缺氧性脑损伤所致的认知功能障碍，其机制可能是通过激活PI3K-Akt通路，上调Bcl-2、下调Bax与活化型胱天蛋白酶3的表达，进而抑制神经元凋亡。
Abstract：Objective To investigate the protective effect and anti-apoptotic mechanism of Buyang Huanwu Decoction(BHD) against acute high-altitude hypoxic brain injury in a mouse model via the PI3K/Akt signaling pathway. Methods 90 male C57 BL/6 mice were randomly assigned to the normal control(NC) group, hypoxic model(M) group, low-/medium-/high-dose BHD(BHD-L, BHD-M and BHD-H) groups, and dexamethasone(Dex) group. Except for the NC group, all mice received oral gavage with respective treatments for four days. On day 5, mice in all groups except NC group were placed in a hypobaric chamber simulating 6 000 m altitude for 72 h to establish the acute high-altitude hypoxic brain injury model. Cognitive function was assessed using the Intelli Cage automated monitoring system. Brain tissue morphology and mitochondrial ultrastructure were examined by HE staining and transmission electron microscopy. Hippocampal neuronal counts were evaluated via Nissl staining, and neuronal apoptosis was detected using TUNEL assay. Serum levels of S100B protein and glial fibrillary acidic protein(GFAP) were measured by ELISA. The protein levels of p-PI3K, p-Akt, Bcl-2, Bax, and cleaved caspase-3 were determined by Western blot. The mRNA levels of PI3K, Akt, Bcl-2, Bax, and Caspase-3 were measured by RT-qPCR. Results In the behavioral tests(nose-poke learning phase, spatial reversal phase, and novel object phase), the M group showed decreased nose-poke times and visits compared to the NC group(P<0.01), indicating impaired cognitive function. Morphological results revealed structural damage to hippocampal neurons, fragmented mitochondrial cristae, and a significantly increased apoptosis rate(P<0.001), accompanied with elevated serum levels of S100B protein and GFAP(P<0.01). Molecularly, the M group showed downregulated protein and mRNA levels of p-PI3K, p-Akt and Bcl-2, while the expression of Bax and cleaved caspase-3 was upregulated(P<0.05). Behavioral tests showed that the BHD-H group and the Dex group had increased visits and nose-poke times compared to the M group(P<0.05). Indicators were restored across all BHD dose groups, with the BHD-H group showing the most significant improvement(P<0.05), and comparable efficacy to Dex(P>0.05). Conclusion Buyang Huanwu Decoction can effectively mitigate cognitive impairment and neuronal damage associated with acute high-altitude hypoxic brain injury in mice. Its neuroprotective effects may be attributed to activation of the PI3K-Akt signaling pathway, leading to enhanced Bcl-2 expression and suppression of Bax and cleaved caspase-3, thereby inhibiting neuronal apoptosis.
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基本信息:
DOI：10.16289/j.cnki.1002-0837.2026.03001
中图分类号:R285.5
引用信息:
[1]朱智芳,郭雪靖,黄珊珊,等.基于PI3K-Akt通路调控海马神经元细胞凋亡探讨补阳还五汤对急性高原缺氧性脑损伤的保护机制[J].航天医学与医学工程().DOI:10.16289/j.cnki.1002-0837.2026.03001.
基金信息:
青海省“昆仑英才·高层次教育教学人才”项目; 青海省中藏医药科研课题项目（J2023003）
2026-03-27
2026-03-27
2026-03-27