The regulatory effect of Xiaoyaosan on the polarization of hippocampal microglia in depressive model mice
10.3760/cma.j.cn371468-20240815-00369
- VernacularTitle:逍遥散对抑郁模型小鼠海马小胶质细胞极化的调节作用
- Author:
Zilong LUO
1
;
Luoyi KUANG
;
Tiezhu YANG
;
Gaoyuan LIU
;
Qing JIA
;
Minzhi WANG
;
Yan LIU
Author Information
1. 河南中医药大学中医学院（仲景学院），郑州 450046
- Publication Type:Journal Article
- Keywords:
Depression;
Xiaoyaosan;
Hippocampus;
Neuroinflammation;
Microglia polarization;
Mouse
- From:
Chinese Journal of Behavioral Medicine and Brain Science
2025;34(5):388-396
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the relationship between the polarization state of hippocampal microglia and depression-like behavior in mice and the regulatory effect of Xiaoyaosan.Methods:Sixty female BALB/C mice were randomly divided into control group, model group, Xiaoyaosan group and fluoxetine group according to the random number method with 15 in each group. Except for control group, the mice in the other 3 groups received chronic restraint stress for 21 days to establish the depressive model. The mice in the Xiaoyaosan group and fluoxetine group were gavaged with Xiaoyaosan(28.06 g/kg) and fluoxetine(3.03 mg/kg) respectively, while the mice in control and model groups received the same volume of 0.9% NaCl solution. Mouse behaviors were evaluated by sucrose preference test and elevated plus maze test.ELISA was used to detect the contents of 5-hydroxytryptamine(5-HT), dopamine(DA) in serum and transforming growth factor-β1(TGF-β1), interleukin-10(IL-10), tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) in the hippocampus. Immunohistochemistry was used to detect inducible nitric oxide synthase(iNOS) and arginase-1(Arg-1) expression.Western blot was used to detect the protein levels of iNOS, Arg-1, CD86 and CD206. The mRNA expression of iNOS and Arg-1 in hippocampus was detected by qRT-PCR. The pathological changes of hippocampus were observed by HE staining.SPSS 27.0 software was used to analyze the data. One-way ANOVA was used for comparison among multiple groups, and LSD test was used for pairwise comparison.Results:(1) There were significant differences in sucrose preference among the 4 groups ( F=46.62, P<0.05).The sucrose preference of model group was lower than that of the control group ( P<0.05), while the sucrose preference of Xiaoyaosan group was higher than that of the model group ( P<0.05).There were significant differences in the number of open arm entries and residence time in the elevated plus maze test among the 4 groups ( F=24.63, 26.94, both P<0.05). The number of open arm entries and residence time in model group ((1.80±1.48)times, (6.19±1.67)s) were lower than those of the control group ((9.80±1.64)times, (56.81±2.75)s)) (both P<0.05). The number of open arm entries and residence time in Xiaoyaosan group ((6.80±0.84)times, (29.59±7.72)s) were significantly higher than model group(both P<0.05).(2) There were significant differences in serum 5-HT and DA levels among the 4 groups ( F=33.27, 76.03, both P<0.05). The serum 5-HT and DA levels in the model group were lower than those of the control group (both P<0.05).The serum 5-HT and DA levels in the Xiaoyaosan group were higher than those of model group (both P<0.05).(3)There were significant differences in the contents of TGF-β1, IL-10, TNF-α and IL-6 in the hippocampus of 4 groups ( F=31.93, 64.01, 25.74, 28.14, all P<0.05). The contents of TGF-β1 and IL-10 in the model group were lower than those of the control group, while the contents of TNF-α and IL-6 were higher (all P<0.05). Compared with the model group, the contents of TGF-β1 and IL-10 in Xiaoyaosan group ( TGF-β1: (30.40±1.56)pg/mL vs (23.77±2.24) pg/mL; IL-10: ((233.94±11.38)pg/mL) vs (130.46±15.34) pg/mL) were higher, and the contents of TNF-α ((73.35±1.51)ng/mL vs (85.89±4.52)pg/mL) and IL-6 (66.15±2.96)pg/mL vs (76.01±1.59)pg/mL) )were lower (all P<0.05).(4)The results of qRT-PCR, immunohistochemistry and Western blot all showed that there were significant differences in mRNA and protein levels of iNOS ( F=41.92, 20.78, 9.27, all P<0.05) and Arg-1 ( F=27.24, 24.23, 6.49, all P<0.05) in the hippocampus among the 4 groups of mice. The mRNA and protein levels of iNOS of the model group were higher than those in the control group (both P<0.05), while the mRNA and protein levels of Arg-1 were lower than those in the control group(both P<0.05).The mRNA and protein levels of iNOS in the Xiaoyaosan group were lower than those in the model group (both P<0.05), while the mRNA and protein levels of Arg-1 were higher than model group(both P<0.05).(5) The expressions of CD206 and CD86 in hippocampus of the 4 groups were significantly different ( F=86.14, 24.02, both P<0.05). Compared with the control group, the model group had a higher expression of CD86 in the hippocampus and a lower expression of CD206 (both P<0.05).Compared with the model group, the CD86 of Xiaoyaosan group was lower, while CD206 was higher (both P<0.05). (6) The HE staining results showed that the cells in the hippocampal CA1 region of the model group mice exhibited disordered arrangement, fewer cells, larger intercellular space, unclear boundary and other changes.The morphology of the cells in the Xiaoyaosan group was improved compared to the model group. Conclusion:Xiaoyaosan can inhibit M1 activation of microglia and neuronal damage in the hippocampus of mice caused by chronic restraint stress, exerting neuroprotective effects and improving depressive behavior in mice.
