Zhang Xin-Rong, Wang Tian-Lang, Zhang Jia-Hao, Jin Lu, Wang Jian-Bo, Xue Ya-Nan, Qu Yi
Liaoning University of Traditional Chinese Medicine Shenyang 110847, China.
Liaoning University of Traditional Chinese Medicine Shenyang 110847, China Key Laboratory of Ministry of Education for Traditional Chinese Medicine Viscera-State Theory and Applications,Liaoning University of Traditional Chinese Medicine Shenyang 110847, China National and Local Joint Engineering Laboratory for Integrated Chinese and Western Medicine Prevention and Treatment Technology on Cardio-Brain Diseases, Liaoning University of Traditional Chinese Medicine Shenyang 110847, China.
Zhongguo Zhong Yao Za Zhi. 2025 Jul;50(14):3857-3867. doi: 10.19540/j.cnki.cjcmm.20250416.501.
This study aimed to investigate the effect and mechanism of Buyang Huanwu Decoction in regulating endoplasmic reticulum stress via the inositol-requiring enzyme 1α(IRE1α)/apoptosis signal-regulating kinase 1(ASK1)/c-Jun N-terminal kinase(JNK) pathway to improve neurological function in rats with cerebral ischemia/reperfusion injury(CIRI). SPF-grade male sprague-dawley(SD) rats were randomly divided into Sham group, model group, Buyang Huanwu Decoction group, and edaravone group. Except for the Sham group, the other groups were subjected to the modified suture method to establish a middle cerebral artery occlusion/reperfusion(MCAO/R) model. After treatment, neurological function was assessed using the Zea Longa scoring system. Gait analysis was used to detect the motor function. Detection of relative infarct area in brain tissue using 2,3,5-triphenyltetrazolium chloride(TTC) staining. Nissl staining was used to observe the structure of neuronal cells. Western blot and real-time fluorescence quantitative PCR(RT-qPCR) were used to detect IRE1α, ASK1, JNK, B cell lymphoma-2(Bcl-2), Bcl-2 related X protein(Bax), and Caspase-3 in the brain tissue. Immunohistochemistry was used to detect the positive expression of IRE1α, ASK1, and JNK. Immunofluorescence was used to detect the fluorescence expression levels of Bax, Bcl-2, and Caspase-3. The results showed that compared with the Sham group, the model group exhibited increased neurological scores(P<0.01), increased ratio of ground contact area and strength in both forelimbs(P<0.01), enlarged relative infarct area of brain tissue(P<0.05), and a reduced number of Nissl staining-positive cells(P<0.01). The protein and mRNA expression levels of IRE1α, ASK1, JNK, Bax, and Caspase-3 in brain tissue were significantly elevated, while those of Bcl-2 were decreased(P<0.05). Compared with the model group, both the Buyang Huanwu Decoction group and edaravone group showed reduced neurological scores(P<0.05), decreased ratio of ground contact area and strength in both forelimbs(P<0.05), smaller relative infarct area(P<0.05), alleviated neuronal damage, and increased number of Nissl staining-positive cells(P<0.05). The expression levels of IRE1α, ASK1, JNK, Bax, and Caspase-3 protein and mRNA in brain tissue were significantly reduced, while those of Bcl-2 were significantly increased(P<0.05). The results indicated that Buyang Huanwu Decoction can effectively improve brain injury in CIRI rats, and its mechanism of action may be related to regulating the endoplasmic reticulum stress IRE1α/ASK1/JNK signaling pathway.
本研究旨在探讨补阳还五汤通过肌醇需求酶1α（IRE1α）/凋亡信号调节激酶1（ASK1）/c-Jun氨基末端激酶（JNK）通路调控内质网应激以改善脑缺血/再灌注损伤（CIRI）大鼠神经功能的作用及机制。将SPF级雄性Sprague-Dawley（SD）大鼠随机分为假手术组、模型组、补阳还五汤组和依达拉奉组。除假手术组外，其他各组采用改良缝线法建立大脑中动脉闭塞/再灌注（MCAO/R）模型。治疗后，采用Zea Longa评分系统评估神经功能。步态分析用于检测运动功能。采用2,3,5-三苯基氯化四氮唑（TTC）染色检测脑组织相对梗死面积。尼氏染色用于观察神经元细胞结构。采用蛋白质免疫印迹法和实时荧光定量PCR（RT-qPCR）检测脑组织中IRE1α、ASK1、JNK、B细胞淋巴瘤-2（Bcl-2）、Bcl-2相关X蛋白（Bax）和半胱天冬酶-3（Caspase-3）的表达。免疫组织化学法检测IRE1α、ASK1和JNK的阳性表达。免疫荧光法检测Bax、Bcl-2和Caspase-3的荧光表达水平。结果显示，与假手术组相比，模型组神经功能评分升高（P<0.01），前肢着地面积和力量比值增加（P<0.01），脑组织相对梗死面积增大（P<0.05），尼氏染色阳性细胞数量减少（P<0.01）。脑组织中IRE1α、ASK1、JNK、Bax和Caspase-3的蛋白和mRNA表达水平显著升高，而Bcl-2的表达水平降低（P<0.05）。与模型组相比，补阳还五汤组和依达拉奉组神经功能评分均降低（P<0.05），前肢着地面积和力量比值降低（P<0.05），相对梗死面积减小（P<0.05），神经元损伤减轻，尼氏染色阳性细胞数量增加（P<重要的是，在与模型组比较时，两个治疗组在这些指标上均有显著改善，表明补阳还五汤和依达拉奉对改善CIRI大鼠的神经功能和脑损伤具有相似的效果。然而，补阳还五汤在调节内质网应激相关信号通路方面可能具有独特的作用机制，值得进一步深入研究。05）。脑组织中IRE1α、ASK1、JNK、Bax和Caspase-3蛋白及mRNA表达水平显著降低，而Bcl-2表达水平显著升高（P<0.05）。结果表明，补阳还五汤可有效改善CIRI大鼠脑损伤，其作用机制可能与调节内质网应激IRE1α/ASK1/JNK信号通路有关。
