Journal of Traditional Chinese Medicine ›› 2024, Vol. 44 ›› Issue (4): 670-679.DOI: 10.19852/j.cnki.jtcm.20240626.002
• Original articles • Previous Articles Next Articles
LIU Tongtong1, ZHANG Xi2, YANG Hui3, LIN Xiaoyuan3, LIU Jian3, ZHANG Xiuli1, GUO Dongwei2, ZHAO Hongqing1, ZOU Manshu1, LEI Chang1, LONG Hongping3, LUO Yan1, XIANG Yun1, GE Jinwen4, WANG Yuhong1, MENG Pan1()
Received:
2023-01-12
Accepted:
2023-08-22
Online:
2024-08-15
Published:
2024-06-26
Contact:
MENG Pan, Institute of Innovation and Applied Research in Chinese Medicine, Hunan University of Chinese Medicine, Changsha 410208, China. Supported by:
LIU Tongtong, ZHANG Xi, YANG Hui, LIN Xiaoyuan, LIU Jian, ZHANG Xiuli, GUO Dongwei, ZHAO Hongqing, ZOU Manshu, LEI Chang, LONG Hongping, LUO Yan, XIANG Yun, GE Jinwen, WANG Yuhong, MENG Pan. Luteolin promotes neuronogenesis in hippocampus of chronic unpredictable mild stress rats and primary hippocampus of fetal rats[J]. Journal of Traditional Chinese Medicine, 2024, 44(4): 670-679.
Group | n | Open field | Latency to feed (s) | Sugar preference (%) | |
---|---|---|---|---|---|
Horizontal | Vertical | ||||
Control | 8 | 114.5±9.4 | 9.6±1.6 | 10.6±10.6 | 77.2±3.5 |
CUMS | 8 | 10.0±2.7a | 1.7±0.5a | 90.7±19.4a | 28.1±3.6a |
Luteolin-H | 8 | 97.6±19.1b | 7.9±1.7b | 7.2±4.0b | 70.9±3.0b |
Luteolin-L | 8 | 16.1±1.7 | 4.4±1.3 | 45.4±14.6 | 36.1±2.2 |
Fluoxetine | 8 | 77.4±7.0b | 6.9±0.9b | 10.9±7.4b | 68.7±3.2b |
Table 1 Behavior scores in each group
Group | n | Open field | Latency to feed (s) | Sugar preference (%) | |
---|---|---|---|---|---|
Horizontal | Vertical | ||||
Control | 8 | 114.5±9.4 | 9.6±1.6 | 10.6±10.6 | 77.2±3.5 |
CUMS | 8 | 10.0±2.7a | 1.7±0.5a | 90.7±19.4a | 28.1±3.6a |
Luteolin-H | 8 | 97.6±19.1b | 7.9±1.7b | 7.2±4.0b | 70.9±3.0b |
Luteolin-L | 8 | 16.1±1.7 | 4.4±1.3 | 45.4±14.6 | 36.1±2.2 |
Fluoxetine | 8 | 77.4±7.0b | 6.9±0.9b | 10.9±7.4b | 68.7±3.2b |
Group | n | 1 week | 2 weeks | 3 weeks | 4 weeks |
---|---|---|---|---|---|
Control | 8 | 214.1±2.5 | 274.2±3.3 | 333.0±5.1 | 404.5±5.4 |
CUMS | 8 | 217.5±2.5 | 240.1±4.6a | 272.6±4.7a | 301.5±3.8a |
Luteolin-H | 8 | 209.2±4.1 | 247.8±2.7 | 290.6±4.8b | 346.4±7.0c |
Luteolin-L | 8 | 216.9±3.4 | 241.6±4.8 | 280.6±6.2 | 310.1±10.5 |
Fluoxetine | 8 | 215.9±4.4 | 244.9±6.1 | 301.8±9.3c | 342.8±4.1c |
Table 2 Body weight in each group (g)
Group | n | 1 week | 2 weeks | 3 weeks | 4 weeks |
---|---|---|---|---|---|
Control | 8 | 214.1±2.5 | 274.2±3.3 | 333.0±5.1 | 404.5±5.4 |
CUMS | 8 | 217.5±2.5 | 240.1±4.6a | 272.6±4.7a | 301.5±3.8a |
Luteolin-H | 8 | 209.2±4.1 | 247.8±2.7 | 290.6±4.8b | 346.4±7.0c |
Luteolin-L | 8 | 216.9±3.4 | 241.6±4.8 | 280.6±6.2 | 310.1±10.5 |
Fluoxetine | 8 | 215.9±4.4 | 244.9±6.1 | 301.8±9.3c | 342.8±4.1c |
Figure 1 Escape latency and crossing distance of the platform position in each group A: the results of Morris Water maze test of each group; B: trace plot of escape latency of control (B1), CUMS (B2), luteolin-H (B3), luteolin-L (B4), fluoxetine (B5) groups.; C: trace plot of Crossing distance of the platform position of control (C1), CUMS (C2), luteolin-H (C3), luteolin-L (C4), fluoxetine (C5) groups. Control: unstressed and intragastrically given 2 mL distilled water daily. CUMS: were intragastrically with distilled water. Luteolin-H: were intragastrically with luteolin at 20 mg·kg-1·d-1. Luteolin-L: were intragastrically with luteolin at 10 mg·kg-1·d-1. Fluoxetine: were intragastrically with fluoxetine at 5.4 mg·kg-1·d-1. CUMS: chronic unpredictable mild stress. Statistical significance among groups was conducted using one-way analysis of variance. Data were presented as mean ± standard error of mean (n = 8). aP < 0.0,1 compared with the control group; bP < 0.05, cP < 0.01, compared with the CUMS group.
Group | n | BDNF | 5-HT | NE | DA |
---|---|---|---|---|---|
Control | 8 | 38.3±1.9 | 75.8±2.0 | 22.5±1.1 | 45.2±0.8 |
CUMS | 8 | 17.5±0.7a | 35.6±3.0a | 13.2±0.7a | 20.5±0.3a |
Luteolin-H | 8 | 32.6±1.9b | 63.5±4.9b | 22.9±1.2b | 45.3±0.6b |
Luteolin-L | 8 | 19.4±0.8 | 46.8±3.2c | 13.9±0.8 | 19.9±0.2 |
Fluoxetine | 8 | 33.9±2.1b | 65.2±4.3d | 24.4±1.0b | 45.8±0.7b |
Table 3 Content of BDNF, 5-HT, NE and DA in serum in each group (ng/mL)
Group | n | BDNF | 5-HT | NE | DA |
---|---|---|---|---|---|
Control | 8 | 38.3±1.9 | 75.8±2.0 | 22.5±1.1 | 45.2±0.8 |
CUMS | 8 | 17.5±0.7a | 35.6±3.0a | 13.2±0.7a | 20.5±0.3a |
Luteolin-H | 8 | 32.6±1.9b | 63.5±4.9b | 22.9±1.2b | 45.3±0.6b |
Luteolin-L | 8 | 19.4±0.8 | 46.8±3.2c | 13.9±0.8 | 19.9±0.2 |
Fluoxetine | 8 | 33.9±2.1b | 65.2±4.3d | 24.4±1.0b | 45.8±0.7b |
Figure 2 Expression of BDNF, NT-3, NGF in the dentate gyrus of hippocampus A: gray value of BDNF; B: gray value of NT-3; C: gray value of NGF. Control: unstressed and intragastrically given 2 mL distilled water daily. CUMS: were intragastrically with distilled water. Luteolin-H: were intragastrically with luteolin at 20 mg·kg-1·d-1. Luteolin-L: were intragastrically with luteolin at 10 mg·kg-1·d-1. Fluoxetine: were intragastrically with fluoxetine at 5.4 mg·kg-1·d-1. CUMS: chronic unpredictable mild stress; BDNF: brain derived neurotrophic factor; NT-3: neurotrophin-3; NGF: nerve growth factor. Statistical significance among groups was conducted using one-way analysis of variance. Data are expressed as mean ± standard error of mean (n = 3). aP < 0.01, compared with the control group; bP < 0.01, compared with the CUMS group.
Figure 3 Neuronal differentiation and migration along the septo-temporal axis of hippocampus in each group Magnification of pictures is 100 ×. A: Representative pictures of BrdU+/NeuN+ cells. A1: expression of BrdU in the S1; A2: expression of NeuN in the S1; A3: expression of DAPI in the S1; A4: merged expression of BrdU, NeuN, and DAPI in the S1; A5: expression of BrdU in the S2; A6: expression of NeuN in the S2; A7: expression of DAPI in the S2; A8: merged expression of BrdU, NeuN, and DAPI in the S2; A9: expression of BrdU in the T3; A10: expression of NeuN in the T3; A11: expression of DAPI in the T3; A12: merged expression of BrdU, NeuN, and DAPI in the T3; A13: expression of BrdU in the T4; A14: expression of NeuN in the T4; A15: expression of DAPI in the T4; A16: merged expression of BrdU, NeuN, and DAPI in the T4; B: BrdU+/ NeuN+ cells per mm3. C: representative pictures of BrdU+/ DCX+ cells. C1: expression of BrdU in the S1; C2: expression of DCX in the S1; C3: expression of DAPI in the S1; C4: merged expression of BrdU, DCX, and DAPI in the S1; C5: expression of BrdU in the S2; C6: expression of DCX in the S2; C7: expression of DAPI in the S2; C8: merged expression of BrdU, DCX, and DAPI in the S2; C9: expression of BrdU in the T3; C10: expression of DCX in the T3; C11: expression of DAPI in the T3; C12: merged expression of BrdU, DCX, and DAPI in the T3; C13: expression of BrdU in the T4; C14: expression of DCX in the T4; C15: expression of DAPI in the T4; C16: merged expression of BrdU, DCX, and DAPI in the T4; D: BrdU+/ DCX+ cells per mm3. Control: unstressed and intragastrically given 2 mL distilled water daily. CUMS: were intragastrically with distilled water. Luteolin-H: were intragastrically with luteolin at 20 mg·kg-1·d-1. Luteolin-L: were intragastrically with luteolin at 10 mg·kg-1·d-1. Fluoxetine: were intragastrically with fluoxetine at 5.4 mg·kg-1·d-1. CUMS: chronic unpredictable mild stress; BrdU: 5-bromo-2-deoxyuridine; DAPI: 4’,6-Diamidino-2’-phenylindole; NeuN: Neuron specific nuclear protein antigen; DCX: doublecortin; S1: septal; S2: mid-septal; T3: mid-temporal; T4: temporal. Statistical significance among groups was conducted using one-way analysis of variance. Data were shown as mean ± standard error of mean (n = 3). aP < 0.01, dP < 0.05, compared with the control group; bP < 0.05, cP < 0.01, compared with the CUMS group.
Group | n | Average fluorescence intensity of BDNF | |||
---|---|---|---|---|---|
S1 | S2 | T3 | T4 | ||
Control | 8 | 0.403±0.023 | 0.600±0.023 | 0.577±0.023 | 0.233±0.022 |
CUMS | 8 | 0.273±0.023a | 0.297±0.020c | 0.213±0.032c | 0.117±0.018a |
Luteolin-H | 8 | 0.383±0.020b | 0.507±0.018d | 0.440±0.021d | 0.190±0.012 |
Luteolin-L | 8 | 0.217±0.015 | 0.337±0.026 | 0.277±0.027 | 0.153±0.015 |
Fluoxetine | 8 | 0.403±0.030b | 0.540±0.044d | 0.477±0.023d | 0.183±0.019 |
Table 4 Expression of BDNF along the septo-temporal axis of hippocampus in the indicated groups
Group | n | Average fluorescence intensity of BDNF | |||
---|---|---|---|---|---|
S1 | S2 | T3 | T4 | ||
Control | 8 | 0.403±0.023 | 0.600±0.023 | 0.577±0.023 | 0.233±0.022 |
CUMS | 8 | 0.273±0.023a | 0.297±0.020c | 0.213±0.032c | 0.117±0.018a |
Luteolin-H | 8 | 0.383±0.020b | 0.507±0.018d | 0.440±0.021d | 0.190±0.012 |
Luteolin-L | 8 | 0.217±0.015 | 0.337±0.026 | 0.277±0.027 | 0.153±0.015 |
Fluoxetine | 8 | 0.403±0.030b | 0.540±0.044d | 0.477±0.023d | 0.183±0.019 |
Group | n | S1 | S2 | T3 | T4 |
---|---|---|---|---|---|
Control | 8 | 0.450±0.012 | 0.593±0.026 | 0.553±0.020 | 0.280±0.017 |
CUMS | 8 | 0.183±0.020a | 0.293±0.015a | 0.207±0.029a | 0.197±0.018d |
Luteolin-H | 8 | 0.277±0.020b | 0.493±0.015c | 0.493±0.026c | 0.213±0.015 |
Luteolin-L | 8 | 0.227±0.015 | 0.313±0.020 | 0.327±0.035 | 0.203±0.015 |
Fluoxetine | 8 | 0.290±0.012b | 0.507±0.035c | 0.463±0.026c | 0.273±0.026 |
Table 5 Expression of NT-3 along the septo-temporal axis of hippocampus in the indicated groups
Group | n | S1 | S2 | T3 | T4 |
---|---|---|---|---|---|
Control | 8 | 0.450±0.012 | 0.593±0.026 | 0.553±0.020 | 0.280±0.017 |
CUMS | 8 | 0.183±0.020a | 0.293±0.015a | 0.207±0.029a | 0.197±0.018d |
Luteolin-H | 8 | 0.277±0.020b | 0.493±0.015c | 0.493±0.026c | 0.213±0.015 |
Luteolin-L | 8 | 0.227±0.015 | 0.313±0.020 | 0.327±0.035 | 0.203±0.015 |
Fluoxetine | 8 | 0.290±0.012b | 0.507±0.035c | 0.463±0.026c | 0.273±0.026 |
Group | n | Average fluorescence intensity of NGF | |||
---|---|---|---|---|---|
S1 | S2 | T3 | T4 | ||
Control | 8 | 0.447±0.020 | 0.550±0.023 | 0.523±0.015 | 0.297±0.020 |
CUMS | 8 | 0.237±0.018a | 0.277±0.020a | 0.293±0.032a | 0.197±0.018a |
Luteolin-H | 8 | 0.277±0.018 | 0.493±0.015b | 0.490±0.015b | 0.263±0.026 |
Luteolin-L | 8 | 0.263±0.009 | 0.343±0.020 | 0.327±0.035 | 0.223±0.015 |
Fluoxetine | 8 | 0.317±0.015 | 0.540±0.038b | 0.487±0.023b | 0.283±0.026 |
Table 6 Expression of NGF along the septo-temporal axis of hippocampus in the indicated groups
Group | n | Average fluorescence intensity of NGF | |||
---|---|---|---|---|---|
S1 | S2 | T3 | T4 | ||
Control | 8 | 0.447±0.020 | 0.550±0.023 | 0.523±0.015 | 0.297±0.020 |
CUMS | 8 | 0.237±0.018a | 0.277±0.020a | 0.293±0.032a | 0.197±0.018a |
Luteolin-H | 8 | 0.277±0.018 | 0.493±0.015b | 0.490±0.015b | 0.263±0.026 |
Luteolin-L | 8 | 0.263±0.009 | 0.343±0.020 | 0.327±0.035 | 0.223±0.015 |
Fluoxetine | 8 | 0.317±0.015 | 0.540±0.038b | 0.487±0.023b | 0.283±0.026 |
Figure 4 Expression of BDNF, NT-3, TrkB, and p-CREB in primary cells in the indicated groups Magnification of pictures is ×400. A: relative integrated density of BDNF; B: relative integrated density of NT-3; C: relative integrated density of TrkB; D: relative integrated density of p-CREB. Control: unstressed and intragastrically given 2 mL distilled water daily. CUMS: were intragastrically with distilled water. Luteolin-H: were intragastrically with luteolin at 20 mg·kg-1·d-1. Luteolin-L: were intragastrically with luteolin at 10 mg·kg-1·d-1. Fluoxetine: were intragastrically with fluoxetine at 5.4 mg·kg-1·d-1. CUMS: chronic unpredictable mild stress; BDNF: brain derived neurotrophic factor; NT-3: neurotrophin-3; TrkB: tropomyosin receptor kinase B; p-CREB: phosphorylated cAMP responsive element binding protein. Statistical significance among groups was conducted using one-way analysis of variance. Data were shown as mean ± standard error of mean (n = 3). aP < 0.01, dP < 0.05, compared with the control group; bP < 0.01, cP < 0.05, compared with the CUMS group.
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