Effects of Chaihu-Shugan San on behavior and hippocampal neurogenesis in depression model mice / 中华行为医学与脑科学杂志

Objective:To explore the effects of Chaihu-Shugan San (CSS) on the behavior and neurogenesis function of depression model mice induced by chronic unpredictable mild stress (CUMS).Methods:Thirty clean grade healthy male C57BL/6 adult mice were randomly divided into control group (Con group), model group (CUMS group) and Chaihu-Shugan San treatment group (CSS group), with 10 mice in each group.The mice in CUMS group and CSS group were given CUMS intervention to establish depression model. At the same time of modeling, the mice in CUMS group and CSS group were given distilled water and CSS(2.7 g/kg) by gavage respectively.While the mice in Con group were only given equal volume distilled water by gavage without CUMS stimulation.After the intervention, the depressive-like behavior of mice was evaluated by increased body weight, sugar water preference test (SPT), forced swimming test (FST) and tail suspension test (TST). The number of newborn neurons was detected by immunofluorescence staining. The mRNA expression levels of brain-derived neurotrophic factor (BDNF), fibroblast growth factor 2 (FGF2) and spindle and kinetochore-associated protein 2(SKA2) in mice hippocampus were detected by qRT-PCR.Statistical analysis was performed by SPSS 22.0 software. One-way ANOVA was used for multi group comparison, and Tukey test was used for pairwise comparison.Results:(1) After modeling, there was significant difference in body weight increment among the three groups ( F=8.859, P <0.05). The body weight increment of CUMS group was lower than those of Con group and CSS group (both P< 0.05). There were significant differences in sugar water preference rate, tail suspension immobility time and swimming immobility time among the three groups ( F=10.544, 12.957, 8.095, all P<0.05). The sugar water preference rate in CUMS group was lower than that in Con group ((87.46±2.78)%, (93.90±3.31)%, P<0.05), and that in CSS group was higher than that in CUMS group ((91.65±2.61)%)( P<0.05). The tail suspension immobility time ((198.00±27.57) s) and swimming immobility time ((322.20±46.98) s) in CUMS group were higher than those in Con group ((138.80±38.50) s, (238.50±50.51) s, both P<0.05). The tail suspension immobility time ((139.00±21.29) s) and swimming immobility time ((265.20±44.90) s) in CSS group were lower than those in CUMS group (both P<0.05). (2) Immunofluorescence showed that there was significant difference in the number of newborn neurons labeled by BrdU and NeuN in the dentate gyrus of hippocampus among the three groups ( F=9.486, P<0.05). The number of double labeled cells (31.66±3.21) in CUMS group was lower than that in Con group(63.66±15.17) and CSS group (58.00±6.00) (both P<0.05). (3) RT-PCR results showed that the mRNA levels of BDNF, FGF2, SKA2 in hippocampal dentate gyrus of the three group were significantly different( F=14.522, 9.337, 8.701, all P<0.05). The levels of BDNF mRNA (0.79±0.06), FGF2 mRNA (0.74±0.18) and SKA2 mRNA (0.52±0.32) in the dentate gyrus of hippocampus in CUMS group were lower than those in Con group (BDNF mRNA (1.03±0.10), FGF2 mRNA (1.04±0.11), SKA2 mRNA (1.05±0.37), all P<0.05). Compared with CUMS group, the mRNA levels of BDNF (1.07±0.80), FGF2 (1.30±0.29) and SKA2 (1.40±0.55) in CSS group were higher (all P<0.05). Conclusion:CSS can alleviate the depressive like behavior of depression model mice, which may be related with increasing the mRNA expression levels of BDNF, FGF2, SKA2 and promoting the proliferation of neural stem cells in hippocampus.

Integrated analysis of brain transcriptome gene expression profile of patients with major depressive disorder / 中华行为医学与脑科学杂志

Objective:To investigate the differential expression of transcriptome genes in various brain regions of depression patients compared with normal people.Methods:Major depressive disorder (MDD) case-control design with transcriptomic studies in gene expression omnibus (GEO) were retrieved and selected in this study, by setting the species as human.To explore the expression distance of brain transcriptome between MDD patients and healthy control subjects, the study first carried out principal component analysis (PCA) based on the variables of the whole genes in each set using network analyst web-based software.Next, differentially expressed gene (DEG) analysis was performed using Limma algorithm and the cut-off values were set as 1.2, 1.5, 2.0-fold changes, along with P<0.05 was considered statistically significant.In order to obtain each brain robust DEGs, the study performed Venn analysis to reveal the common DEGs among independent datasets.Last, DAVID online software was used to perform functional enrichment analysis of stable and differentially expressed genes, and explore the changes of GO function and KEGG pathway in different brain regions of patients with depression. Results:Compared with the control group, the whole expression level of transcriptome in each brain region (anterior cingulate gyrus, prefrontal lobe, amygdala) of MDD patients was slightly changed, since PCA plots could not distinguish the health controls from MDD patients based on the whole gene expression level.There were almost no differentially expressed genes with multiple variation more than 2.0-fold changes.The results of Venn analysis indicated that there was no overlappped gene in each independent brain regions.Functional enrichment analysis to these DEGs with fold changes ≥ 1.2 showed that these genes were enriched in some GO terms and KEGG pathways which were not directly associated with MDD, further implying that gene expression changes were not potential forces driving the onset of depression.Conclusion:These results based on the integration of multiple datasets highlight that gene expression levels exhibit a small fluctuation between MDD and normal brains.Thus, it is still challenging to recognize the specific genes of depression susceptibility due to various limitations.

Immune inflammation and major depression disorder / 中华行为医学与脑科学杂志

In recent years,the research direction of the pathogenesis of depression has gradually ex-panded from classical neurotransmitter disorders and neuroendocrine disorders theory to the related fields of immuno-inflammatory response and researchers believe that depression is a psycho-neuro-immune disorder disease. Because many patients with chronic inflammation,cancer and autoimmune diseases are easy to suffer depression,it confirms that the occurrence of depression may be related to the adverse effects of immune in-flammation in the brain. A large number of clinical and experimental studies have shown that the role of im-mune inflammation in depression is mainly due to the release of inflammatory factors from immune cells through the blood-brain barrier,activation or intensification of brain immune cell response,or changes in the structure and function of neuroendocrine axis,neurotransmitters,emotional regulation-related brain regions, leading to the occurrence and development of depression. The purpose of this review is to summarize the pos-sible mechanisms and links of immune inflammation in depression in recent years,and to provide ideas and methods for the research and treatment of depression.

Regulation of Chaihu Shugan Powder on kynurenine in raphe nucleus of depressive rats induced by reserpine / 中华行为医学与脑科学杂志

Objective To investigate the antidepressant effects of Chaihu Shugan Powder(CSP) on depressive rats induced by reserpine and its influences on the kynurenine (KYN),indoleamine 2,3-dioxyge-nase(IDO),interleukin-6(IL-6) and tumor necrosis factor-α( TNF-α). Methods Forty rats with similar behavior results were divide into 4 groups randomly,including Control group(Con),Model group(Res),Flu- oxetine group(Res+Flu) and Chaihu Shugan Powder group(CSP). The depressive rat model was established by intraperitoneal injection reserpine. The rats in Res+Flu group were administered with fluoxetine by intrap-eritoneal and rats in Res+CSP group were administered with CSP by intraperitoneal. After 14 days,the be-havior of rats was measured and then the rats were executed and sampled. The content of tryptophan and kynurenine in raphe nuclei tissue were detected. The mRNA expression level of IDO,IL-6,TNF-α in raphe nuclei tissue were detected. Results ( 1) Compared with Con group (( 81. 81 ± 36. 13) s, ( 83. 51 ± 5. 34)%), the swimming immobility time((150. 50±31. 45)s) in Res group increased(t=68. 7, P<0. 05) and the sucrose perference (59. 73±11. 30)%) in Res group decreased(t=23. 8,P<0. 05). Compared with Res group, the swimming immobility time in Res+Flu group((114. 90± 14. 29) s) and Res+CSP group ((111. 7±11. 34)s) decreased(t=35. 6,35. 8,both P<0. 05). Compared with Res group, the sucrose pref-erence in Res+Flu group((78. 21±10. 07)%) increased(t=18. 3, P<0. 05). (2)Compared with Con group (KYN/TRP:(0. 023±0. 016),IDO mRNA:(1. 00±0. 05),IL-6 mRNA:(1. 00±0. 58),TNF-α mRNA:(1. 00±0. 32)), the activity of IDO(KYN/TRP(0. 039±0. 003)) and the mRNA levels of IDO mRNA(3. 63± 0. 31),IL-6 mRNA(2. 36±0. 23),TNF-α mRNA( 3. 56± 0. 14) of Raphe Nuclei tissue in Res group were significantly increased (t=21. 2,12. 9,38. 3,19. 7,all P<0. 05). Compared with Res group, the activity of IDO(KYN/TRP(0. 030±0. 013)),the mRNA expression levels of IDO mRNA( 1. 56±0. 36),IL-6 mRNA (1. 62±0. 16),TNF-α mRNA(2. 64±0. 20)of Raphe Nuclei tissue in Res+Flu group were significantly de-creased(t=38. 8,15. 8,12. 8,26. 4,all P<0. 05). And compared with Res group,the activity of IDO( KYN/TRP(0. 028±0. 021)) ,the mRNA expression level of IDO mRNA( 1. 33± 0. 29),IL-6 mRNA(1. 36± 0. 34),TNF-α mRNA(1. 93±0. 21)of raphe nuclei tissue in Res+CSP group were also significantly decreased (t=23. 21,17. 3,19. 8,29. 8,all P<0. 05). Compared with Res+Flu group,the level of IDO mRNA and in-flammatory factors' mRNA in Res+CSP group were significantly decreased(t=18. 3,20. 8,31. 5,all P<0. 05). Conclusion Chaihu Shugan Powder has antidepressant effect,and the mechanism is related with de-creasing the inflammatory factors,inhibiting IDO activation and decreasing the IDO mRNA.

Changes of behavioral and serum inflammatory factors in rats with acute reserpine-induced comor-bidity of pain and depression / 中华行为医学与脑科学杂志

Objective To investigate the effects of acute reserpine-induced pain and depression co-morbidity model on behavior and related inflammatory cytokines in rats. Methods Twelve male 8-week-old SD rats were randomly divided into control group and model group,6 in each group. Rats in the model group were intraperitoneally injected with reserpine(1 mg/kg/d)for 3 days to establish the model. Rats in the con-trol group were injected with the same amount of distilled water. The pain threshold of rats was measured by Von Frey Hairs and Hot Plate Analgesia Test. The depression behavior of rats was evaluated by open-field test,Forced Swimming Test and Sucrose Consumption Test. Serum IL-1β, IL-6, IL-18 and TNF-α content were detected by ELISA. Results Compared with the control group(1d (15. 00±0. 00) g;3d ( 13. 20±4. 03)g),the PWTs of the model group (1d (6. 20±0. 45) g;3d (4. 20±1. 64) g) decreased significantly(t=44. 00,4. 63,both P<0. 05). Compared with the control group (( 8. 82 ± 1. 08) s),the PWTL (( 3. 16 ± 0. 24)s) was significantly reduced on the first day in model group,and the difference was statistically signifi-cant (t=11. 48,P<0. 05). Compared with the control group (( 1 815. 18± 541. 40) cm,(98. 20± 26. 25) s, (87. 78±9. 38)g),the total distance of the open-field test ((948. 91±494. 35)cm)significantly shortened (t=2. 64,P<0. 05),swimming time ((143. 60±21. 45)s) was significantly prolonged (t=-2. 65,P<0. 05), and the sucrose consumption (( 22. 23 ± 6. 97) g) significantly decreased (t=12. 55,P<0. 05) in model group. Compared with the control group (( 285. 80 ± 11. 93) ng/ml, ( 233. 07 ± 8. 47) ng/ml,( 280. 41 ± 14. 31) ng/ml,( 213. 10 ± 33. 87) ng/ml), serum levels of IL-1β, IL-6, IL-18, TNF-α in model group ((471. 23±24. 15) ng/ml,(364. 82±17. 16) ng/ml,(471. 81± 28. 98) ng/ml,(821. 19±93. 16) ng/ml) increased significantly(F=-15. 39,-15. 39,-13. 24,-13. 72,all P<0. 05). Conclusion A large number of intraperitoneal injections of acute reserpine can cause hyperalgesia,depressive behavior and serum inflamma-tory factors in rats,effectively simulating the symptoms of pain and depression,and can be used as a model of pain and depression comorbidity for biological mechanisms and treatment research.

Research Progress in Pharmacological Effects and Mechanism of Chaihu Shugan Powder / 中国中医药信息杂志

Chaihu Shugan Powder has the function of soothing liver and promoting qi flow, activating blood and relieving pain. In clinic, it is mainly used for the syndrome of liver qi stagnation, with wide application. This article reviewed the literature about effects of Chaihu Shugan Powder on neurobiochemistry, endocrine regulation, immunity and antioxidant pharmacological effects and mechanism, with a purpose to provide references for clinical medication and new medicine research.

Effect of Chaihu Shugan Powder on the content of 5-hydroxytryptamine and the expression of tryptophan hydroxylase 2 in depression rats / 中华行为医学与脑科学杂志

Objective To explore the effect of Chaihu Shugan Powder (CSP) on the content of 5-hydroxytryptamine (5-HT) in the hippocampus and the expression of tryptophan hydroxylase 2(TPH2) in median raphe nuclei in depression rats.Methods SD rats were randomly divided into normal group(n=12),model group (n=12),fluoxetine group (n=12),low-dose and high-dose CSP group (n =12,respectively).Depression model was made by reserpine intraperitoneal injection.During the experiment,the weight and the open-field scores were calculated;the content of 5-HT was detected by ELISA.The expression of TPH2 in median raphe nuclei was detected by Western blot.Results Compared with the weight ((225.02±5.23) g),the open-field scores ((12.6± 5.1)score) and content of 5-HT ((1.09±0.27) ng/ml) in the model group,high-dose CSP showed significantly improve the depressive rats in weight,open field score and content of 5-HT ((238.78±5.16) g,(15.6±7.8) score and (1.80±0.58) ng/ml,respectively;P＜0.05 or P＜0.01).The expression of TPH2 (0.66±0.21) in median raphe nuclei in the high-dose CSP group was apparently increased compared with that in the model group(0.16±0.04) (P ＜0.01).Conclusion CSP have the effects of anti-depression,which could be related with the increase of the 5-HT content in the hippocampus and the expression of TPH2 in median raphe nuclei.

Effect of Fuzhenghuayu compound on hepatocyte expression of Nrf2 in a mouse model of hepatic fibrosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of Fuzhenghuayu compound (FZHc) on expression of nuclear factor E2-related factor 2 (Nrf2) in hepatocytes under conditions of hepatic fibrosis using a mouse model.</p><p><b>METHODS</b>Mice were randomly assigned to a control group and a hepatic fibrosis model group. The control group was further divided into three subgroups for use as normal controls (A1), mineral oil-treated controls (A2), and FZHc-treated controls (A3); the hepatic fibrosis model group was administered carbon tetrachloride (CC14 dissolved in mineral oil and injected intraperitoneally) and further divided into four subgroups for use as 6-weeks models (B1), 10-weeks models (B2), low-dose (L)-FZHc models (C1), and high-dose (H)-FZHc models (C2). The FZHc (capsule powder diluted with double-distilled water to 0.1 g/mL) was administered via gastric perfusion to groups A3, C1, and C2 starting at week 7 of the experiment. At the end of week 6 and 10, hepatic specimens were collected and evaluated for degree of hepatic fibrosis and inflammation using routine haematoxylin-eosin staining and Masson staining. Immunohistochemical analysis was performed to measure the hepatocyte expression of Nrf2, NAD(P)H quinine oxidoreductase 1 (Nqol), a-smooth muscle actin (a-SMA) and fibronectin (FN). Real-time fluorescence quantitative PCR was used to measure Nrf2 mRNA expression. Western blotting was used to detect Nrf2 and Nqol total protein expression and Nrf2 nuclear translocation. F test, LSD test and ridit test were used for statistical analyses.</p><p><b>RESULTS</b>Compared with the B2 group (ridit value: 0.09), the model groups treated with FZHc showed significantly lower degrees of hepatic inflammation and fibrosis for both the low (C1 group, ridit value: 0.32) and high doses (C2 group, ridit value: 0.40) (F =82.927, P less than 0.05). In addition, compared with the B2 group, the model groups treated with FZHc showed significantly decreased expression of a-SMA and FN proteins, with a dose-dependent trend (by immunohistochemistry: C 1 group at the end of 10 weeks, F =77.421, 118.262, P less than 0.05; C2 group, P =0.002, 0.013) and significantly increased expression of Nrf2 and Nqol proteins (by immunohistochemistry:C1 and C2 groups at the end of 10 weeks, F =182.537, 75.615, P less than 0.05 and by westen blotting: F =45.664, 127.673, P less than 0.05), which also showed a dose-dependent trend (C2 group, P =0.000, 0.014; 0.005, 0.014). Western blotting also indicated that the amount of nuclear transported Nrf2 was higher in the C1 and C2 groups at the end of 10 weeks (vs. B2 group, F =94.787, P less than 0.05), and the amount of nuclear transported Nrf2 was significantly higher in the C2 group (vs. C1 group, P =0.044). Nrf2 mRNA expression was significantly higher in the C1 group than in the B2 group (F =3230.105, P less than 0.05), and the C2 group had more substantially increased expression (P =0.001); there was no statistical difference found between groups B1 and B2 (P =0.094).</p><p><b>CONCLUSION</b>Fuzhenghuayu compound increased the expression of Nrf2 mRNA and protein under conditions of hepatic fibrosis in mice and stimulated Nrf2 nuclear transport, as well as increased expression of the Nrf2 target gene Nqol that is known to suppress activation of hepatic stellate cells and decrease the deposition of FN. Therefore, Fuzhenghuayu compound may ameliorate hepatocyte injury in hepatic fibrosis in mice by exerting an antihepatic fibrosis effect.</p>

Electrophysiological study of BDNF gene-modified mesenchymal stem cell transplantation to repair transversely hemisectioned spinal cord injury in rats / 中国实验动物学报

Objective To study the effects of bone marrow mesenchymal stem cells modified by brain-derived neuro-trophic factor ( BDNF) gene on the repair of spinal cord injury by electrophysiological assay .Methods Thirty healthy Spra-gue-Dawley rats (male and female) were randomly divided into 3 groups:Blank group, 10 rats (removal of the lamina only and exposed spinal dura mater );spinal cord injury (SCI) group,10 rats;and cell transplantation after SCI group , 10 rats. Eight rats of them were selected randomly and detected their SEP and MEP , and evaluated the degree of recovery of motor scores in the rats at 1 d, 7 d, 14 d, 21 d, 30 d, and 60 d.Result Since 4 days after cell transplantation , the process of hind limbs changes was as follows:at the 1-4 days after injury , the injury side hind limb had flaccid paralysis , mopping the floor walk, the movement of contralateral hind limb was gradually recovered from the initial injury , the injury side hind limb had spastic paralysis in 5-9 days after SCI;during 10-14 days, the injury side had a few activities;the contralateral side re-covered to a less normal state;At 15-21 days, activities of the injury side improved obviously , until the 30th day.The activ-ity and muscle tension degree of the injury side recovered most obviously .After 30 days no more obvious improvement was ob-served.Immunohistochemistry showed that the transplanted mesenchymal stem cells , which were induced and labeled firstly , survived at the damage spinal cord , and behavioral observation found that the cell transplantation improved exercise capacity of the rats injured before .Conclusion Bone marrow mesenchymal stem cells modified by BDNF gene can partially promote the recovery of nerve transmission function and nerve regeneration .

Relationship between Lp-PLA_2 and vulnerable atherosclerotic plaque in rabbits / 中国病理生理杂志

AIM: To explore the expressive role of lipoprotein-associated phospholipase A_2, high sensitive C-reactive protein and matrix metalloproteinase-9 in vulnerable atherosclerotic plaques in a rabbit model. METHODS: Forty eight New Zealand white male rabbits were randomly divided into 4 groups (12 rabbits each): control group, stable plaque group, p53 group, and p53+drug group. Rabbits in control group were fed with a regular diet and underwent sham operation. Rabbits in stable plaque group, p53 group and p53+drug group underwent balloon induced arterial wall injury and then were fed on a diet with 1% cholesterol. The animals were all fed for 3 months, then the rabbits in p53 group and p53+drug group underwent Ad5-CMV p53 transfection at 10th week. Before killed, the animals in p53+drug group underwent pharmacological triggering with Russell's viper venom (RVV) and histamine to induce the rupture of the atherosclerotic plaques. At the 1st day and before sacrifice, the serum was collected for measuring Lp-PLA_2, hs-CRP, MMP-9, HDL, LDL and VLDL. The expressions of Lp-PLA_2, hs-CRP and MMP-9 in tissues were determined by the methods of hybridization and immunohistochemistry. RESULTS: At the end of 12th week, the serum and tissue levels of Lp-PLA_2 and MMP-9 in stable plaque group, p53 group and p53+drug group were significant different from those in control group and in each group at the first day (P<0.05). The serum levels of Lp-PLA_2 and hs-CRP in p53 group and p53+drug group were significantly higher than those in control group and stable group (P<0.05). The serum levels of Lp-PLA_2, hs-CRP and MMP-9 were all significantly different between p53 group and p53+drug group (P<0.05). At the end of 12th week, pathological results showed that 4 groups were normal artery, stable plaque, vulnerable plaque and rupture plaque, respectively. The fabric cap was thicker in plaque groups than that in normal group (P<0.05). The rupture and formation of thrombus were more significant in p53+drug group than those in p53 group. The serum level of Lp-PLA_2 had negative interrelated relationship with fabric cap in plaque groups (r=-0.710, P<0.01), and hs-CRP, MMP-9 had no interrelated relationships with fabric cap in plaque groups. CONCLUSION: Base on the successful establishment of the atherosclerotic plaque animal model, serum Lp-PLA_2 shows better interrelated relationships to plaques stability. Combination with hs-CRP and MMP-9, we can exactly evaluate the nature of plaques.

Regulation of glucocorticoid receptor and its mRNA by Tsiao Shihutang / 中国病理生理杂志

AIM: In order to explore the mechanism of the GC - like effect of Tsiao Shaihutang (TSS), the down - regulation of glucocorticoid receptor and mRNA by TSS were studied. METHODS: GR sites were determined by receptor radio ligand binding assay method. At the same time, GR mRNA level was determined by quantitative reverse- transcriptive polymerase chain reaction (RT- PCR) method. RESULTS: (1) GR sites and GR mRNA level were down - regulated significantly after GC, TSS (P < 0.01 ) treatment . (2) GR sites and GR mRNA level in GC plus TSS group were obviously higher than those in GC group (P < 0.01 ). CONCLUSIONS: These data suggested that TSS can significantly down- regulate GR and GR mRNA level.