Chaihu-Shugan-San Group Research Articles

Effect of Chaihu-Shugan-San on functional dyspepsia and gut microbiota: A randomized, double-blind, placebo-controlled trial

Ethnopharmacological relevanceChaihu-Shugan-San (CSS) is a classic traditional Chinese medicine (TCM) formula from the Ming Dynasty "Jingyue's Complete Works”. In China, it is prevalent for the treatment of a wide range of ailments, with a particular emphasis on functional gastrointestinal disorders (FGIDs). Clinical evidence suggests that CSS has been found to be a highly effective therapeutic approach for the treatment of Functional Dyspepsia (FD), however, there is a limited amount of high-quality clinical evidence, particularly randomized, double-blind, placebo-controlled trials to support this claim. Aim of the studyTo evaluate the therapeutic efficacy of Chaihu-Shugan-San (CSS) for treating functional dyspepsia (FD) by comparing it to placebos, as well as to investigate the impact of CSS on the gut microbiota in individuals diagnosed with FD. Materials and methodsThis was a randomized double-blind, placebo-controlled clinical trial implemented at Shuguang Hospital in Shanghai. Between May 2021 and December 2022, 94 participants satisfying the Rome IV diagnostic criteria for FD were enrolled. They were assigned randomly to either the CSS group or the placebo group, with an equal allocation ratio of 1:1. Patients in both groups received the intervention for four weeks. The primary outcome was the dyspepsia symptom scores evaluated by using single dyspepsia symptom scale (SDS) after four weeks of treatment. The secondary outcomes were the solid gastric empties rate measured by a barium strip method, Hamilton anxiety scale (HAMA), Hamilton depression scale (HAMD), and Functional dyspepsia Quality of life scale (FDDQL). In addition, after unblinding, 30 patients in the CSS group were randomly selected and divided into before and after treatment of the FD groups (FD1, FD2), and 30 healthy participants were selected as healthy control group (HC), and the gut microbiota was analyzed by 16S rRNA sequencing. ResultsAfter four weeks of treatment, the SDS score exhibited a significant improvement in the CSS group compared to the placebo group (t = 4.882; P ＜0.001). The difference in barium strip gastric emptying rate in the CSS group showed a significant ascent compared to the control group (P < 0.01). The HAMA, HAMD, and FDDQL scores in the CSS group showed a statistically significant increase compared to the control group (all P < 0.01). The results of 16S rRNA sequencing revealed that FD patients had less diverse and abundant microbiota than the healthy people. Additionally, the application of CSS resulted in the modulation of certain bacterial populations, leading to both up-regulation and down-regulation of their quantities. ConclusionsThese findings suggested that CSS is more effective compared to a placebo in treating FD, relieves anxiety and depression, increases gastric emptying rate in FD patients, and that CSS also affects the bacterial community structure in FD patients. Trial registrationChiCTR, ChiCTR2100045793. Registered 25 Mach 2021. URL: https://www.chictr.org.cn/showproj.html?proj=125049.

Antidepressant-Like Effects and Cognitive Enhancement of Coadministration of Chaihu Shugan San and Fluoxetine: Dependent on the BDNF-ERK-CREB Signaling Pathway in the Hippocampus and Frontal Cortex.

Background Fluoxetine (FLU) is the first-line and widely used medication for depression; however, FLU treatment is almost ineffective in 30%-40% of patients with depression. In addition, there are some problems in FLU treatment, such as delayed efficacy, large side effects, and poor tolerance. Chaihu Shugan San (CSS) is a classic and effective antidepressant Chinese herbal medicine that has been used in China for thousands of years. CSS or coadministration of CSS and FLU has become one of the most recommended methods in the treatment of depression in China. However, the specific pathways of CSS and coadministration of CSS and FLU for antidepressant are still unclear. Objective This study was designed to evaluate the antidepressant effects of CSS and coadministration of CSS and FLU. Methods The chronic unpredictable mild stress (CUMS) rat model was used to simulate depression. 120 healthy adult male Sprague-Dawley (SD) rats were randomly divided into seven groups: the control group, CUMS group, low-dose CSS group, high-dose CSS group, FLU group, coadministration of low-dose CSS and FLU group, and coadministration of high-dose CSS and FLU group. The rats in different groups were given different interventions. Then, the depression-like behavior and cognitive function were evaluated by the sucrose preference test (SPT), forced swimming test (FST), open field test (OFT), and Y-maze test. What is more, the antidepressant mechanism of CSS and coadministration of CSS and FLU were studied through BDNF mRNA, ERK mRNA, CREB mRNA, BDNF, p-ERK/ERK, and p-CREB/CREB levels in the hippocampus and frontal cortex by Western blot and RT-PCR. Results Compared with the CUMS group, CSS and coadministration of CSS and FLU could alleviate the depressive symptoms and improve cognitive function in CUMS rats (p < 0.05); CSS and coadministration of CSS and FLU could increase the expression of BDNF, p-CREB/CREB, p-ERK/ERK, and BDNF mRNA, CREB mRNA, and ERK mRNA in the hippocampus and frontal cortex (p < 0.05); CSS and coadministration of CSS and FLU could increase the expression of BDNF, p-CREB/CREB, p-ERK/ERK, and BDNF mRNA, CREB mRNA, and ERK mRNA in the hippocampus and frontal cortex (p < 0.05); CSS and coadministration of CSS and FLU could increase the expression of BDNF, p-CREB/CREB, p-ERK/ERK, and BDNF mRNA, CREB mRNA, and ERK mRNA in the hippocampus and frontal cortex (Discussion and Conclusion. Finally, we found that both CSS and coadministration of CSS and FLU play an antidepressant role, which may be due to the regulation of the BDNF/ERK/CREB signaling pathway in the hippocampus and frontal cortex. Among them, the coadministration of CSS and FLU can enhance the antidepressant effect of CSS or FLU alone, and the underlying mechanism needs further investigation.

Effect of chaihu-shugan-san on the behaviors and Notch1 signal pathway in depression model rats

Objective To investigate the effect of chaihu-shugan-san (CSS) on the behaviors and Notch1 signal pathway in depression model rats. Methods Thirty-two SD rats with similar behavioral scores were divided into control group (CON), model group (CUMS), positive control group (FLU) and intervention group (CSS). The depression model was established by stimulating with chronic unpredictable mild stress (CUMS), and the behaviors evaluation was assessed by sugar water consumption and forced depression.Immunofluorescence was used to detect the proliferation of hippocampus neurons in rats, at the same time, real-time PCR and Western blot were used to detect the mRNA and protein expression levels of each factor (Notch1, Hes1, Hes5 and Jagged1) of Notch1 signal pathway respectively. Results Compared with CON group, the percentage of sugar water preference and swimming length of rats decreased significantly in CUMS group (P<0.05 and P<0.01). Compared with CUMS group, the percentage of sugar water preference and swimming length of rats increased significantly in CSS group(P<0.05). Compared with CON group, there was a significant increase in the inactivity length of rats between CUMS group, FLU group and CSS group, and the difference was statistically significant (P<0.01). Compared with CUMS group, the swimming length of rats in CSS group was significantly reduced, and the difference was statistically significant (P<0.01). Compared with CON group((750.00±27.51)/mm2), the number of BrdU positive cells in the substratum or granulocyte layer of the hippocampus dentate gyrus of rats in CUMS group ((338.75±29.61)/mm2), FLU group ((545.00±17.73)/mm2) and CSS group ((529.38±13.74)/mm2) was significantly reduced(P<0.01). Compared with CUMS group ((338.75±29.61)/mm2), there was a significant increase in the number of BrdU positive cells in the substratum or granulocyte layer of the hippocampus dentate gyrus of rats in CSS group ((529.38±13.74)/mm2), and the difference was statistically significant (P<0.01). Compared with CON group, the mRNA and protein expression levels of Notch1, Hes1, Hes5, and Jagged1 in the hippocampus of rats in CUMS group were significantly reduced(P<0.05 or P<0.01). Compared with CUMS group, the mRNA and protein expression levels of Notch1, Hes1, Hes5 and Jagged1 in the hippocampus of rats in FLU group and CSS group were significantly increased, and the differences were statistically significant (P<0.05 or P<0.01). Conclusion Notch1 signal pathway may be related to the obstacle during the hippocampus nerve regenerating in the model rat under chronic unpredictable mild stress.CSS may play an anti-depressant role by regulating Notchl to improve hippocampus nerve regeneration. Key words: Chaihu-shugan-san(CSS); Chronic unpredictable mild stress; Depression; Notch1 signal pathway

Effect of Chaihushugan San on expression of the Raf/mitogen-activated protein kinase/extracellular signal-regulated kinase pathway in the hippocampi of perimenopausal rats induced by immobilization stress

ObjectiveWe wished to study the impact of Chaihushugan San (CSS) on the behavior of perimenopausal rats with liver-Qi stagnation (LQS) and to investigate the effect of CSS on signal transduction of the Raf/mitogen-activated protein kinase (MEK)/extracellular signal-regulated kinase (ERK) cascade in the hippocampi of rats induced by immobilization. MethodsTwenty 52-week-old female rats were divided into two groups by the random number table method: model control group (MCG) and CSS group (CSSG), with 10 rats in each group. Ten-week-old female rats were used as the normal control group (NCG). CSS effects were assessed using rats exposed to immobilization stress by measuring body weight and sucrose consumption, serum hormone levels, and observing performance in the open field test (OFT). Molecular mechanisms were examined by measuring the effect of CSS on expression of Raf1, MEK1/2 and ERK1/2 mRNA in hippocampi using quantitative real-time polymerase chain reaction and by measuring levels of these proteins and related phospho-proteins using Western blotting. ResultsPerimenopausal rats with LQS had decreased locomotor activity; reduced sucrose consumption; and increased serum levels of corticotropin releasing hormone (CRH) and corticosterone (CORT). Activation of hippocampal Raf/MEK/ERK cascade was suppressed significantly in the MCG, and activation was increased after 21 days of CSS treatment. ConclusionCSS has significant effects upon relief of the symptoms of LQS in immobilization-induced rats. The mechanism underlying this action might (at least in part) be mediated by reversal of disruption of the Raf/MEK/ERK pathway.

Changes in regional cerebral blood flow with Chaihu-Shugan-San in the treatment of major depression.

Background:Chaihu-Shugan-San (CHSGS) is a well-known Chinese traditional prescription used for depression.Objective:To observe the regional cerebral blood flow (rCBF) changes in patients with major depression and to investigate rCBF and clinical response to CHSGS.Materials and Methods:A total of 33 unmedicated patients with major depression and 12 healthy comparison subjects underwent single photon emission computed tomography (SPECT) imaging. A total of 33 unmedicated patients with major depression all met the diagnostic criteria of stagnation of liver qi of traditional Chinese medicine and were divided into two groups: CHSGS group (n = 20) and fluoxetine group (n = 13). SPECT imaging was restudied in posttreatment.Results:SPECT detected abnormalities in all (100.0%) patients both in CHSGS group and fluoxetine group. All healthy subjects were normal results. The depressed patients showed rCBF decreased in the multiple regions. The semiquantitative values of bilateral frontal and left temporal lobes both in CHSGS group and fluoxetine group were lower than that in healthy group (P < 0.05). Reexamined SPECT after 8 weeks treatment with CHSGS showed the consistency between the increase in perfusion defects and the improvement of clinical cerebral symptoms. The semiquantitative values increased in posttreatment, when compared with pretreatment (P < 0.05).Conclusion:SPECT represents a sensitive tool to detect the major depressive disorder, which show the rCBF decreased. rCBF perfusion defects can be reversed and clinical symptoms can be improved by CHSGS treatment. CHSGS treatment is effective, well-tolerated, and safe for depression. By semiquantitative analysis, SPECT can objectively detect rCBF changes that is useful for guiding treatment.

Antidepressant-like activity of Chaihu-Shugan-San aqueous extract in rats and its possible mechanism.

Background:Chaihu-Shugan-San (CHSGS), a traditional Chinese medicinal formula, is commonly used for the treatment of depression in China. However, the molecular mechanism underlying its antidepressant action is unknown.Objective:The objective of this study is to evaluate the antidepressant-like effects of CHSGS and further explore the possible molecular mechanism implicated in its actions.Materials and Methods:The rats were randomly divided into four groups: The normal control group, the model control group, the CHSGS group and the fluoxetine control group. The antidepressant-like effects of CHSGS aqueous extract were assessed in rats exposed to chronic mild stress (CMS) using the open-field test and sucrose water consumption test, its underlying mechanism of anti-depression was explored by determining the effect of CHSGS on the extracellular signal-regulated kinase (ERK) and phospho-ERK (P-ERK) in the hippocampus using western blot. The aqueous extract of CHSGS at a dose of standard (5.9 g/kg·d) was administered intragastrically for 14 days during the CMS model while the fluoxetine control group was given at the same time using fluoxetine hydrochloride (1.8 mg/kg·d).Results:The stressed rats demonstrated decreased locomotor activity in open field test and reduction in sucrose consumption and decreased levels of P-ERK1/2 and the ratio of P-ERK1/2 to total ERK1/2 in the hippocampus. CHSGS alleviated the depressive-like behaviors and increased levels of P-ERK1/2 and the ratio of P-ERK1/2 to total ERK1/2 in stressed rats as well as fluoxetine.Conclusion:In summary, these results suggest that CHSGS aqueous extract possesses an antidepressant-like activity in CMS induced depression model rats, which might be mediated, at least in part, by reversing the stress-induced disruption of ERK activity.

P38 Mapk signal pathway involved in anti-inflammatory effect of chaihu-shugan-san and shen-ling-bai-zhu-san on hepatocyte in non-alcoholic steatohepatitis rats

Traditional Chinese Medicine (TCM), has over thousands-of-years history of use. Chaihu-Shugan-San (CSS), and Shen-ling-bai-zhu-San (SLBZS), are famous traditional Chinese herbal medicine formulas, which have been used in China, for the treatment of many chronic diseases. This study investigated the anti-inflammatory effects of CSS and SLBZS on signaling molecules involved in p38 mitogen-activated protein kinase (p38 MAPK), pathway on hepatocytes of non-alcoholic steatohepatitis (NASH), rats induced by high fat diet. SD male rats were randomly divided into 8 groups: negative control group, model control group, high (9.6g/kg/day)/low (3.2g/kg/day)-dose CSS group, high (30g/kg/day)/low (10g/kg/day)-dose SLBZS group, high (39.6g/kg/day)/low (13.2g/kg/day)-dose integrated group. The rats of NASH model were induced by feeding a high-fat diet. After 16, wks, Hepatocytes were isolated from 6, rats in each group by collagenase perfusion. The liver histopathological changes and serum inflammatory cytokines TNF-α, IL-6 were determined. The proteins of TLR4, phosphor-p38 MAPK and p38 MAPK involved in p38 MAPK signal pathway were assayed. The statistical data indicated the NASH model rats reproduced typical histopathological features of NASH in human. CSS and SLBZS ameliorated lipid metabolic disturbance, attenuated NASH progression, decreased the levels of TNF-α and IL-6 in serum, as well as inhibited TLR4 protein expression, p38 MAPK phosphorylation, and activation of p38 MAPK. In conclusion, CSS and SLBZS might work as a significant anti-inflammatory effect on hepatocyte of NASH by inhibiting the activation of TLR4, p-p38 MAPK and p38 MAPK involved in p38 MAPK signal pathway. To some extent, CSS and SLBZS may be a potential alternative and complementary medicine to protect against liver injury, alleviate the inflammation reaction, moderate NASH progression.

Screening of specific microRNA in hippocampus of depression model rats and intervention effect of Chaihu Shugan San

To screen microRNAs with specific expression of in hippocampus of rats with chronic stress induced depression model, and observe the effect of traditional Chinese medicine Chaihu Shugan San on the expression of microRNA in hippocampus. SD rats were randomly divided into 3 groups: the normal control group, the model control group and the Chaihu Shugan San group. The depression model was replicated by unpredictable chronic mild stress combined with separation. Behavioral changes of the rats were observed by Open-field test and sucrose solution consumption test, and the expression of microRNAs in hippocampus was assayed by microRNA micro-array. Compared with the normal control group, there were 13 specific miRNAs in hippocampus in the model control group with the expression difference of more than 2 times. Among them, down-regulating miRNAs included miR298, miR-130b, miR-135a, miR-323, miR-503, miR-15b, miR-532, and miR-125a, and the up-regulation miRNAs included miR7a, miR-212, miR-124, miR-139, and miR-182. Among the 13 specific miRNAs, miR-125a and miR-182 recovered to normal after intervention with Chaihu Shugan San in the Chaihu Shugan San group. This study preliminarily found that 13 specific miRNAs in hippocampus are related to depression. Among them, miR-125a and miR-182 recover to normal after intervention with Chaihu Shugan San, which may be the target points of the antidepressant effect of Chaihu Shugan San. We shall further analyze the target genes and their mechanisms.

Effects of Chaihu Shugan San on behavior and CRHmRNA expression in the brain of mice with chronic mild unpredicted stress depression

Objective To investigate the effects of Chaihu Shugan San(CHSGS) ,a compound traditional Chinese herbal medicine,on the behavior and plasma levels of corticosterone level and corticotrophin releasing hormone ( CRH) mRNA expression in the prefrontal cortex and the hypothalamus of mice with chronic mild unpredicted stress depression. Methods 40 adult male Kunming mice were randomly divided into 4 groups: the normal control group,the model control group,the fluoxetine group and CHSGS group. Except the normal control group, mice were exposed to an unpredicted sequence of mild stressor for continuous 21 days to induce depression. Since the 8th day, mice were intragastrically administered with equal volume agents respectively for 14 days (normal saline for the normal control group and the model group,fluoxetine(2. 6 mg · kg-1 ) for fluoxetine group and CHSGS (8.45 g · kg-1) for CHSGS group). Body weight and behaviors were investigated,the plasma levels of CORT in different groups were detected by radioimmunoassay, and the mRNA expression in the prefrontal cortex and the hypothalamus were semiquantified by the RT-PCR method. Results In comparision with the model group, body weight changes ( (13.22 ±3.57)g) and consumption of sucrose solution((1. 90 ±0. 74) ml) of CHSGS groups were significantly increased, and the duration of immobility during the forced swimming ((62.20±13.73)s) and tail suspension((58. 10 ± 11.71 )s) ,the levels of plasma CORT((12.61 ±4.63)ng/L) and the CRHmRNA expression in the prefrontal cortex(0.34 ±0.10) and the hypothalamus(0.70 ±0.09) were significantly decreased. Conclusion CHSGS can reverse the depressive behavior, reduced the levels of plasma CORT and down-regulated CRHmRNA expression in the prefrontal cortex and the hypothalamus of the depressive mouse caused chronic stress. Key words: Chaihu Shugan San; Stress; Mice; Behavior; Corticotrophin releasing hormone

Effect of Chaihu Shugan San and its components on expression of ERK1/2 mRNA in the hippocampus of rats with chronic mild unpredicted stress depression.

To explore the antidepressant effect and mechanism of Chaihu Shugan San (CHSGS) composition, a compound traditional Chinese herb medicine and its components. Rats were randomly divided into 6 groups: normal control group, model control group, a CHSGS group, a component I group, a component II group and a fluoxetine control group. The depression model was replicated by chronic unpredictable mild stress and single house for 28 d. Behavioral scores of the rats were detected by Open-field test and sucrose solution consumption test, and ERK1/2 mRNA expression in the hippocampus tissue was assayed by real-time fluorescent quantitative PCR. ERK1/2 mRNA expression was down-regulated in the depression model group compared with the normal control group (P<0.01). Compared with the model group, ERK1/2 mRNA expression in the CHSGS group and fluoxetine group was both up-regulated (P<0.05 or P<0.01); and only ERK1 mRNA expression in the component I group was up-regulated (P<0.05). No significant difference existed between the component II group and the model group (P>0.05). Isolated-living condition and chronic mild unpredictable stress can down-regulate the expression of ERK1/2 mRNA in the hippocampus tissue. CHSGS may exert an antidepressant effect through increasing the expression of ERK1/2 mRNA in the hippocampus, component I may play an important role in its antidepression effect, while compatibility of the use of component II can enhance the antidepressant efficacy.