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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.
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ABSTRACT OBJECTIVE：To establish a method for simultaneous determination of 8 components in the lyophilized product of Chaihu shugan san decoction. METHODS：UPLC method was adopted to determine the contents of albiflorin，paeoniflorin，ferulic acid，naringin，hesperidin，benzoyl paeoniflorin，glycyrrhizic acid and α-cyperone in 6 batches of lyophilized product of Chaihu shugan san decoction. The determination was performed on Phenomenex column with mobile phase consisted of 0.1％ formic acid water-acetonitrile at the flow rate of 1 mL/min，the sample volume was 10 μL. The detection wavelength was set at 250 nm，and column temperature was 30 ℃. RESULTS：The linear range of albiflorin，paeoniflorin，ferulic acid，naringin，hesperidin，benzoyl paeoniflorin，glycyrrhizic acid and α-cyperone were 3.606-8.414，23.988-55.972，1.218-2.842，35.964-83.916，12.009-28.021， 1.194-2.786，3.609-8.421，5.294-12.352 μg/mL，respectively（r＝0.999 5-0.999 9）. The limits of quantitation were 0.206，0.178， 0.256，0.168，0.196，0.242，0.268，0.157 μg/mL，respectively. RSDs of precision，stability（12 h）and repeatability tests were all lower than 2％（n＝5 or 6）；the recoveries were 97.93％，98.18％，96.57％，97.61％，98.51％，97.45％，98.14％，96.91％（all RSD＜2％ ）. The average contents of albiflorin， paeoniflorin， ferulic acid， naringin， hesperidin， benzoyl guanosine， and glycyrrhizic acid in 6 batches of samples were 59.258，429.237，23.173，625.847，200.424，15.048，67.620 μg/g，respectively. α-cyperone was not detected because of its volatility. CONCLUSIONS：The method is simple，stable，reliable and accurate. It could provide reference for quality control of the lyophilized product of Chaihu shugan san decoction
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Objective:To observe effect of Xiaoai Jiedu formula combined with fluorouracil (5-FU)+epirubicin (EPI)+cyclophosphamide (CTX) (FEC) chemotherapy regimen on immune function, tumor index, traditional Chinese medicine (TCM) symptom scale score and adverse reactions of patients with breast cancer. Method:A total of 60 patients with breast cancer were randomly divided into study group and control group. FEC (CTX 0.6 g·m-2,d1,EPI 100 mg·m-2,d1,5-FU 0.5 g·m-2,d1) regimen was used in study group, and Xiaoai Jiedu formula recipe was used for two consecutive cycles. FEC regimen was used in control group only. After 2 cycles, immunological changes, tumor index, TCM symptom score and adverse reactions were analyzed and compared between two groups. Result:There was not significant difference in immune indexes between two groups before treatment, but statistically significant differences after treatment (PPPPConclusion:Xiaoai Jiedu formula combined with FEC chemotherapy can improve clinical efficacy and alleviate clinical symptoms of patients.
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<p><b>OBJECTIVE</b>To investigate the protective effects of Chinese medicine formulation Chaihu Shugan San (, CHSGS) on nonalcoholic fatty liver disease (NAFLD) in rats with insulin resistance (IR) and its molecular mechanisms.</p><p><b>METHODS</b>Male Sprague-Dawley rats were randomly divided into six groups: the control group, the model group, Dongbao Gantai group (, DBGT, 0.09 g methionine/kg), CHSGS high-dose group (CHSG-H, 12.6 g crude drug/kg), CHSGS medium-dose group (CHSG-M, 6.3 g crude drug/kg), and CHSGS low-dose group (CHSG-L, 3.15 g crude drug/kg). After establishing the NAFLD rat model and treatment for 8 weeks, total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), free fatty acid (FFA), fasting blood glucose (FBG), fasting insulin (FINS) contents in blood serum, and TC, TG contents in the hepatic homogenate were measured by an automatic biochemical analyzer, and a homeostasis model assessment was applied to assess the status of IR, insulin sensitivity index (ISI), and homeostasis model assessment for insulin secretion (HOMA-IS). The expression levels of adiponectin and leptin mRNA in liver tissue were analyzed by reverse transcription polymerase chain reaction. Pathological changes of livers were observed by hematoxylin-eosin staining of paraffin section.</p><p><b>RESULTS</b>Compared with the model group, the serum levels of TC, TG, FFA, FBG, FINS, IRI, ISI, and the liver levels of TC and TG in CHSG-H, CHSG-M, CHSG-L groups showed significant declines (P<0.01 or P<0.05); the serum levels of HDL-C, HOMA-IS were significantly increased (P<0.01 or P<0.05); the expression of leptin mRNA was dramatically decreased and the expression of adiponectin mRNA was increased in the hepatic tissue (P<0.01 or P<0.05). The fatty deposition of liver cells could also be alleviated.</p><p><b>CONCLUSION</b>CHSGS could up-regulate the expression of adiponectin mRNA and down-regulate the expression of leptin mRNA on the liver, suggesting the CHSGS had positive therapeutic effect on NAFLD in rats with IR.</p>
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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 interven-tion 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.Im-munofluorescence 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 lay-er 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 ex-pression 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). Conclu-sion 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.
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Objective To evaluate the intestinal absorption of main components (paeoniflorin,ferulic acid,narirutin,naringin,neohesperidin,and glycyrrhizic acid) in aqueous extract of Chaihu Shugan San (CSS).Methods UPLC method was established to simultaneously determine the concentrations of the six components in the test samples of everted gut sacs and in situ single pass intestinal perfusion (SPIP) models.Absorption kinetics parameters were calculated for describing their absorption characteristics.Results In SPIP model,the results indicated that ferulic acid was the well-absorbed ingredient in whole small intestine,while other ingredients presented moderate or poor absorption.In everted gut sacs model,paeoniflorin in jejunum,ferulic acid,naringin,and neohesperidin in duodenum,narirutin in duodenum and jejunum,had the best absorption,while there was no significant difference in absorption of glycyrrhizic acid in the intestine.Condusion In vivo model indicated that the main constituents in CSS could be absorbed in intestinal wall of rat,ferulic acid could be much more easily penetrated intestinal wall into the blood circulation than the other five components.Ex vivo model could further articulate that six index components could be absorbed selectively in different intestinal segments.
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Objective To observe the effects of stress treatment for different time on autonomic behaviors and spatial learning-memory abilities of rats and to investigate the therapeutic efficacy of Chaihu Shugan San(CSS). Methods Seventy-five 8-week-old Wistar rats were randomly divided into blank control group, model control group, and low-, middle-, and high- dose CSS groups, 15 rats in each group. Chronic multi-stress model was set up. The rat body weight in various groups was measured. The autonomic behaviors of rats were observed by open field test, and the abilities of spatial learning-memory of rats were examined by Morris water maze test. Results (1)On modeling day 20, 30, the model control group had lighter body weight than the blank control group (P 0.05). (2)Open field test results showed as follows:Compared with the blank control group, the total distance and central-area distance were shortened (P 0.05). Conclusion The chronic stress may decrease the excitability, reduce autonomic behaviors, and injure spatial learning and memory abilities of rats, the effect being positively correlated with the modeling time. And CSS has an effect on improving the above indexes, in particular the middle-and high-dose of CSS.
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Objective To evaluate the intestinal absorption of main components (paeoniflorin,ferulic acid,narirutin,naringin,neohesperidin,and glycyrrhizic acid) in aqueous extract of Chaihu Shugan San (CSS).Methods UPLC method was established to simultaneously determine the concentrations of the six components in the test samples of everted gut sacs and in situ single pass intestinal perfusion (SPIP) models.Absorption kinetics parameters were calculated for describing their absorption characteristics.Results In SPIP model,the results indicated that ferulic acid was the well-absorbed ingredient in whole small intestine,while other ingredients presented moderate or poor absorption.In everted gut sacs model,paeoniflorin in jejunum,ferulic acid,naringin,and neohesperidin in duodenum,narirutin in duodenum and jejunum,had the best absorption,while there was no significant difference in absorption of glycyrrhizic acid in the intestine.Condusion In vivo model indicated that the main constituents in CSS could be absorbed in intestinal wall of rat,ferulic acid could be much more easily penetrated intestinal wall into the blood circulation than the other five components.Ex vivo model could further articulate that six index components could be absorbed selectively in different intestinal segments.
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Objective To explore the antidepressant effect and mechanism of Chaihu Shugan San (CHSGS) composition, a compound traditional Chinese herb medicine and its components. Methods 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. Results 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). Conclusion 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.
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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.