Tang Ying-Juan, Guo Hai-Peng, Zou Man-Shu, Han Yuan-Shan, Liu Jun-Cheng, Wang Yu-Hong
Technology Innovation Center, Hunan University of Chinese Medicine Changsha 410208, China.
Technology Innovation Center, Hunan University of Chinese Medicine Changsha 410208, China Hunan Key Laboratory of Traditional Chinese Medicine Prevention and Treatment of Depressive Diseases Changsha 410208, China.
Zhongguo Zhong Yao Za Zhi. 2025 Jan;50(1):1-9. doi: 10.19540/j.cnki.cjcmm.20240712.705.
This study investigated the mechanism of Chaishao Kaiyu Decoction in improving hippocampal neuroinflammation in depressed rats based on complement component 3(C3)/C3 receptor(C3aR). A total of 60 SD rats were randomly divided into a blank group, a model group, high, medium, and low dose groups of Chaishao Kaiyu Decoction, and a positive drug group, with 10 rats in each group. Except for the blank group, chronic unpredictable mild stress(CUMS) was used to construct depression models in other groups. Sucrose preference, open-field experiment, forced swimming, and water maze were used to detect the changes in depression-like behavior in each group. Enzyme-linked immunosorbent assay(ELISA) was used to detect the serum inflammatory factor level in rats, and hematoxylin-eosin(HE) staining and Nissl staining were employed to observe the pathological damage of hippocampal neurons. Golgi-Cox staining was used to observe the dendritic spine damage of hippocampal neurons, and immunofluorescence and Western blot were utilized to detect the expression of microglial marker Iba-1 and C3/C3aR protein in the hippocampus of rats. The behavioral results showed that compared with the model group, Chaishao Kaiyu Decoction could significantly strengthen the sugar water preference, increase the distance and number of voluntary activities, shorten the immobility time in forced swimming and the successful incubation period of positioning navigation, and prolong the stay time of space exploration in the target quadrant. ELISA results showed that the content of inflammatory factors in the hippocampus of depressed rats was significantly higher than that of the blank group, and the content of inflammatory factors decreased significantly after the intervention of Chaishao Kaiyu Decoction. In addition, Chaishao Kaiyu Decoction could relieve pathological damage such as cell swelling and loose arrangement of hippocampus tissue. In the Western blot experiment, the expression levels of C3 and C3aR proteins in the model group were higher than those in the blank group, while the expression of C3 and C3aR in Chaishao Kaiyu Decoction could be down-regulated. Immunofluorescence results showed that compared with the model group, the fluorescence intensity of microglia marker Iba-1 decreased significantly after the intervention of Chaishao Kaiyu Decoction and positive drugs. The antidepressant effect of Chaishao Kaiyu Decoction may be related to the down-regulation of C3/C3aR signaling pathway-related proteins, thus alleviating hippocampal inflammation.
本研究基于补体成分3（C3）/C3受体（C3aR）探讨柴芍开郁汤改善抑郁大鼠海马神经炎症的机制。将60只SD大鼠随机分为空白组、模型组、柴芍开郁汤高、中、低剂量组及阳性药物组，每组10只。除空白组外，其他组采用慢性不可预测温和应激（CUMS）构建抑郁模型。采用蔗糖偏好试验、旷场实验、强迫游泳实验和水迷宫实验检测各组抑郁样行为的变化。采用酶联免疫吸附测定（ELISA）检测大鼠血清炎症因子水平，采用苏木精-伊红（HE）染色和尼氏染色观察海马神经元的病理损伤。采用高尔基-考克斯染色观察海马神经元树突棘损伤情况，采用免疫荧光和蛋白质印迹法检测大鼠海马中微胶质细胞标志物Iba-1及C3/C3aR蛋白的表达。行为学结果显示，与模型组相比，柴芍开郁汤可显著增强糖水偏好，增加自主活动距离和次数，缩短强迫游泳不动时间及定位航行成功潜伏期，延长目标象限空间探索停留时间。ELISA结果显示，抑郁大鼠海马中炎症因子含量显著高于空白组，柴芍开郁汤干预后炎症因子含量显著降低。此外，柴芍开郁汤可减轻海马组织细胞肿胀、排列疏松等病理损伤。蛋白质印迹实验中，模型组C3和C3aR蛋白表达水平高于空白组，而柴芍开郁汤可下调C3和C3aR的表达。免疫荧光结果显示，与模型组相比，柴芍开郁汤及阳性药物干预后微胶质细胞标志物Iba-1荧光强度显著降低。柴芍开郁汤的抗抑郁作用可能与下调C3/C3aR信号通路相关蛋白有关，从而减轻海马炎症。
