Background: Chai Hu has the hepato-protective, choleretic, anti-tussive, analgesic, anti-inflammatory, anti-viral, hypotensive, hypolipidemic, and anti-tumor pharmacological effects. In this study, the objective of this paper was to study the anti-depressant effect of total saikosaponins. 
Materials and Methods: Total saikosaponins were extracted by reflux method, and were identified by thin layer chromatography (TLC). The anti-depressant effect of total saikosaponins was investigated in vitro by tail suspension test, forced swimming test, and reserpine antagonism test in mice. 
Results: Two times of reflux extraction, temperature of 70℃, and extraction time of 4hrs, for each extraction could improve the yield of saikosaponins. Each treatment group (100, 200, and 300 mg/kg), could significantly shorten the immobility time of mice in the tail suspension test in a somewhat dose-dependent manner. The total saikosaponins antagonized the reserpine-induced akinesia, and ptosis in mice. 
Conclusion: Total saikosaponins have an anti-depressant effect

Cao, Chunmei

Ding, Haijun

Liu, Yu

African Journal of Traditional Complementary and Alternative Medicines

PubMed

Background: Chai Hu has the hepato-protective, choleretic, anti-tussive, analgesic, anti-inflammatory, anti-viral, hypotensive, hypolipidemic, and anti-tumor pharmacological effects. In this study, the objective of this paper was to study the anti-depressant effect of total saikosaponins.    Materials and Methods: Total saikosaponins were extracted by reflux method, and were identified by thin layer chromatography (TLC). The anti-depressant effect of total saikosaponins was investigated in vitro by tail suspension test, forced swimming test, and reserpine antagonism test in mice.    Results: Two times of reflux extraction, temperature of 70 degrees C, and extraction time of 4hrs, for each extraction could improve the yield of saikosaponins. Each treatment group (100, 200, and 300 mg/kg), could significantly shorten the immobility time of mice in the tail suspension test in a somewhat dose-dependent manner. The total saikosaponins antagonized the reserpine-induced akinesia, and ptosis in mice.    Conclusion: Total saikosaponins have an anti-depressant effect.Integrative &amp; Complementary MedicineSCI(E)02280-2841

Institutional Repository of Peking University

PHARMACOLOGICAL EXPERIMENTAL STUDY OF THE ANTI-DEPRESSANT EFFECT OF   TOTAL SAIKOSAPONINS

Background: Chai Hu has the hepato-protective, choleretic, anti-tussive, analgesic, anti-inflammatory, anti-viral, hypotensive, hypolipidemic, and anti-tumor pharmacological effects. In this study, the objective of this paper was to study the anti-depressant effect of total saikosaponins.Materials and Methods: Total saikosaponins were extracted by reflux method, and were identified by thin layer chromatography (TLC). The anti-depressant effect of total saikosaponins was investigated in vitro by tail suspension test, forced swimming test, and reserpine antagonism test in mice.Results: Two times of reflux extraction, temperature of 70℃, and extraction time of 4hrs, for each extraction could improve the yield of saikosaponins. Each treatment group (100, 200, and 300 mg/kg), could significantly shorten the immobility time of mice in the tail suspension test in a somewhat dose-dependent manner. The total saikosaponins antagonized the reserpine-induced akinesia, and ptosis in mice.Conclusion: Total saikosaponins have an anti-depressant effect.Key words: total saikosaponins; tail suspension test; forced swimming tes

Liu, Y

Cao, C

Ding, H

AJOL - African Journals Online

Liu et al., Afr J Tradit Complement Altern Med. (2014) 11(2):280-284http://dx.doi.org/10.4314/ajtcam.v11i2.9280PHARMACOLOGICAL EXPERIMENTAL STUDY OF THE ANTI-DEPRESSANT EFFECT OF TOTALSAIKOSAPONINS.Yu Liu1, Chunmei Cao2*, Haijun Ding31Department of Psychology, Peking University, Beijing 100871, China. 2Department of Physical Education, TsinghuaUniversity, Beijing 100084, China. 3Fusong Hospital of Traditional Chinese Medicine, Fusong 134500, China*E-mail: pingkew32@126.comAbstractBackground: Chai Hu has the hepato-protective, choleretic, anti-tussive, analgesic, anti-inflammatory, anti-viral, hypotensive, hypolipidemic,and anti-tumor pharmacological effects. In this study, the objective of this paper was to study the anti-depressant effect of total saikosaponins.Materials and Methods: Total saikosaponins were extracted by reflux method, and were identified by thin layer chromatography (TLC). Theanti-depressant effect of total saikosaponins was investigated in vitro by tail suspension test, forced swimming test, and reserpine antagonism testin mice.Results: Two times of reflux extraction, temperature of 70℃, and extraction time of 4hrs, for each extraction could improve the yield ofsaikosaponins. Each treatment group (100, 200, and 300 mg/kg), could significantly shorten the immobility time of mice in the tail suspension testin a somewhat dose-dependent manner. The total saikosaponins antagonized the reserpine-induced akinesia, and ptosis in mice.Conclusion: Total saikosaponins have an anti-depressant effect.Key words: total saikosaponins; tail suspension test; forced swimming testIntroductionChai Hu, which was formerly known as Zi Hu, was originally recorded in the "Shen Nong's Herbal Classic" as a medium-grade drug. Itwas renamed in the Song Dynasty to Chai Hu in the "Maps of Materia Medica". Chai Hu comes from Bupleurum chinensis DC. or Bupleurumscorzonerifolium, of family Umbelliferae; the former is named "Bei Chai Hu", and the latter named "Nan Chai Hu" according to the medicationhabits. Chai Hu is bitter in taste, slightly cold, enters the liver and gallbladder meridians. It has the effects of releasing exterior and curing fever,soothing liver and relieving liver depression, and ascending yang qi; and used in the treatment of alternating episodes of chills and fever, chestfullness, rib pain, bitter taste in the mouth, deafness, headache, dizziness and other symptoms (Liu et al., 2006). Scholars at home and abroad haveconducted in-depth researches on the chemical constituents of Chai Hu, and have isolated saponins, flavonoids, lignans, coumarins,phenylpropanols, polyacetylenes, volatile oils, polysaccharides, polyols, fatty acids, sterols and amino acids from different parts of Chai Hu.Modern pharmacological studies have shown that Chai Hu has the hepatoprotective, choleretic, antitussive, analgesic, anti-inflammatory,anti-viral, hypotensive, hypolipidemic and anti-tumor pharmacological effects (Zhang., 2000; Liu et al., 2002; Shao.,2002; Wang et al., 2002; Zhuet al., 2005) . In this study, total saikosaponins were extracted and separated from Chai Hu and identified, and the anti-depressant effect andmechanism of saikosaponins investigated.Materials and MethodsReagents and InstrumentsProzac capsules (Eli Lilly Suzhou Pharmaceutical Co., Ltd.), reserpine (Guangdong Bangmin Pharmaceutical Co., Ltd.]; TC-15 thermostat(Wuxi Shengze Physical and Chemical Instruments Co., Ltd.); circulating water multipurpose vacuum pump (Tianjin Zhaoyuan VacuumEquipment Manufacturing Co., Ltd.); far-infrared drying oven (Shanghai Pudong Rongfeng Scientific Instrument Co., Ltd.); tail suspensiondevice, mouse rectal temperature probe.AnimalsKunming mice, male, 18-22g, were purchased from the China Medical University, and adaptively fed for four days. All experimentalprocedures were approved by the Animal Research Ethics Committee.Liu et al., Afr J Tradit Complement Altern Med. (2014) 11(2):280-284http://dx.doi.org/10.4314/ajtcam.v11i2.9281Preparation and identification of total saikosaponinsPreparation of total saikosaponins500g of Bei Chai Hu was weighed out, ground into coarse powder, added with methanol containing 5% pyridine, and soaked overnight,then extracted twice under reflux, reflux temperature was 70℃, and reflux time was 4hrs, for each extraction, followed by filtration. The solventwas then recovered to obtain a concentrated solution. The concentrated solution was diluted, and extracted by addition of saturated n-butanol for atotal of three times, the extracts were combined, recovered, and the resulting residue was the total extract containing saikosaponins andsapogenins. The total extract was then added with adequate amount of ether and extracted under reflux for 30min., the ether insoluble matter wascollected, and dissolved by adding appropriate amount of hot ethyl acetate, then ethyl acetate was evaporated, and the residue was dried, andcrushed to obtain total saikosaponins.TLC identification of saikosaponinsAppropriate amount of the above total saikosaponins was weighed out, and dissolved in methanol as the test solution. TLC test wasperformed, developing conditions were: ethyl acetate-ethanol-water (8:2:1), the plates were developed, taken out, air dried, sprayed with 2%solution of p-dimethylaminobenzaldehyde in 40% sulfuric acid, and heated at 60℃ till well-defined pink spots were produced, which indicatedthe presence of total saikosaponins.Tail suspension test in miceThe mice were randomly divided into five different groups; namely the blank control group, fluoxetine group (7 mg/kg), and saikosaponingroups (100, 200, and 300 mg/kg). The mice were intra-gastrically administered for seven consecutive days, 1hr, after the last administration, thetail of mice 2cm from the tip was fixed on a horizontal surface, so that the mice were hung upside down, and the immobility time of mice within6min., was recorded.Forced swimming test in miceAfter the pre-test, the mice were placed in a 15cm deep bucket, the observation began after the mice had swum for 2min., and theimmobility time of mice within 4min., was accumulated, that is, the duration of time in which mice stops struggling, with only small movementsto remain not submerged under water, mice with the immobility time exceeding 120sec., or less than 60sec., were removed from the test. Thequalified mice were randomized, and intra-gastrically administered for seven consecutive days, 1hr, after the last administration, forcedswimming test was performed according to the pre-test methods, and the immobility time of mice within 4min., was accumulated.Reserpine antagonism test (Zheng et al., 2005)1hr, after the last administration, 4mg/kg reserpine was injected intra-peritoneally to the mice in each experimental group, 4hr, later,electronic thermometer probe was inserted approximately 1cm, into the rectum of mice to measure rectal temperature for 10sec., and differencesin the change of rectal temperature of mice in each group before and after 4hrs, were recorded. 1hr, after the intra-peritoneal injection of reserpine,the mice were placed in a circle 8cm, in diameter and observed for 20sec., the out-of-circle rate of mice in each group was calculated, in order toobserve the akinetic state of mice in each group. 1hr, after the intra-peritoneal injection of reserpine, the number of mice with ptosis was counted,and the percentage was calculated.Statistical analysisExperimental data were processed using SPSS 11.5, statistical software, and were expressed as x—±s; comparison among groups wasperformed by the t-test.Liu et al., Afr J Tradit Complement Altern Med. (2014) 11(2):280-284http://dx.doi.org/10.4314/ajtcam.v11i2.9282ResultsEffect of saikosaponins on tail suspension test in miceThe experimental results showed that each experimental group could significantly shorten the immobility time of mice in the tailsuspension test (P<0.01 or P<0.05), in a somewhat dose-dependent manner, as shown in Figure 1.Figure 1: Effect of saikosaponins on tail suspension test in miceEffect of saikosaponins on forced swimming test in miceAs can be seen from the data, compared with the blank control group, the Prozac group could significantly shorten the immobility time ofmice, the saikosaponin medium- and high-dose groups, had similar effects as well, but the saikosaponin low-dose group had no statisticallysignificant effect (see Table 1).Table 1: Effect of saikosaponins on immobility time of mice in the forced swimming test (x—±s, n=10)Group Dose (mg/kg) Immobility time (s)Blank control group 95.2±35.6Prozac group 7 53.4±23.8*Saikosaponin group (low) 100 86.4±30.9Saikosaponin group (medium) 200 56.8±35.5*Saikosaponin group (high) 300 46.3±29.1**Note: Comparison with the blank control group, * P<0.05, ** P<0.01.Reserpine antagonism testIn the reserpine-induced akinesia, and ptosis antagonism test in mice, the saikosaponin medium- and high-dose groups antagonized thereserpine-induced akinesia and ptosis in mice; the antagonistic effect was not obvious in the low-dose group. The three dose groups could allantagonize the reserpine-induced hypothermia in mice (Tables 2, 3 and Figure 2)Table 2: Antagonistic effect of saikosaponins on reserpine-induced ptosis in mice (x—±s, n=12)Group Dose (mg/kg) Number of mice with ptosis Ptosis percentage (%)Blank control group 0 0Reserpine group 4 11 91.67Saikosaponin group (low) 100 8 66.67Saikosaponin group (medium) 200 5 41.67*Saikosaponin group (high) 300 4 33.33*Note: Comparison with the reserpine group, * P<0.05Liu et al., Afr J Tradit Complement Altern Med. (2014) 11(2):280-284http://dx.doi.org/10.4314/ajtcam.v11i2.9283Table 3: Antagonistic effect of saikosaponins on reserpine-induced akinesia in mice (x—±s, n=12)Group Dose (mg/kg) Number of mice moving out of the circle Out-of-circle rate (%)Blank control group 0 100.00Reserpine group 4 1 8.33Saikosaponin group (low) 100 8 66.67Saikosaponin group (medium) 200 6 50.00*Saikosaponin group (high) 300 4 33.33*Note: Comparison with the reserpine group, * P<0.05Figure 2: Effect of saikosaponins on reserpine-induced hypothermia in miceDiscussionChai Hu has a complex chemical composition, its main constituents are saponins and volatile oils, and it also contains flavonoids, fattyacids, polyols, sterols, lignans, coumarins, polysaccharides, trace elements and other types of constituents. Scholars at home and abroad havedone extensive researches on saikosaponins, and have isolated more than 100, kinds of saponin compounds from it; of which the contents ofsaikosaponin d, a, c were relatively high (Luo et al., 1996; Ebata et al., 1990; Liang et al., 1998; Jin et al., 1996; Ebata et al., 1996). Saikosaponinsd, a, c, are all thermally unstable substances, changes occur easily during their extraction and separation process, so the extraction time andtemperature should be strictly limited. It can be seen through the analysis of preliminary experimental results that the extraction conditions of twotimes of reflux extraction, temperature of 70℃, and extraction time of 4hrs, for each extraction can effectively reduce the degradation ofsaikosaponins, and improve its yield.Depression is a mood disorder caused by one or multiple reasons which is mainly characterized by low mood, emotional lowness is itscore symptom, and its main clinical manifestations are despondency, reduced verbal and physical actions, and retardation of thinking. Depressioncan be classified into primary depression and secondary depression, and can generally be divided into exogenous depression, endogenousdepression and bipolar disorder depression (Thase et al., 2009; Filakovic etal., 2009).The mouse models of forced swimming and tail suspension used in this study are sensitive to most anti-depressant drugs, and are widelyused in the preliminary screening of these drugs.Our experimental results showed that each treatment group (100, 200, and 300 mg/kg), could significantly shorten the immobility time ofmice in the tail suspension test in a somewhat dose-dependent manner. In the reserpine-induced akinesia and ptosis antagonism test in mice, thesaikosaponin medium- and high-dose groups could antagonize the reserpine-induced akinesia and ptosis in mice; the antagonistic effect was notobvious in the low-dose group.AcknowledgementThe work was supported by Research on the impact of different load and unsteady on the coordination of body trunk and lower limb,Independent scientific research plan, Ministry of Education, 2011.11-2014.12.Liu et al., Afr J Tradit Complement Altern Med. (2014) 11(2):280-284http://dx.doi.org/10.4314/ajtcam.v11i2.9284References1. Ebata, N., Nakajima, K., Taguchi, H., and Mitsuhashi, H. (1990). Isolation of New Saponins From the Root of Bupleurum falacatum L.Chemical and Pharmaceutical Bulletin 38(5): 1432-1433.2. Ebata N. (1996). Saponins from the Root of Bupleurum falacatum. Phytochemistry, 41(3): 895-901.3. Filakovic, P., Petek, A., Koic, O., Radanovi-Grguri, L., and Degmeci, D. (2009). Comorbidity of Depressive and DermatologicDisorders-therapeutic Aspects. Psychiatria Danubina. 21(3): 401-410.4. Jin, H. F., Jiang, Y., and Luo, S. Q. (1996). Chemical constituents of Bupleurum longicaule var. franchetii and root of Bupleurum chaishouiShan et Sheh. China Journal of Chinese Materia Medica, 21(12): 739-741.5. Liang, H., Zhao, Y. Y., and Qiu, H. Y. (1998). Structural Identification of a New Saikosaponin From Bupleurum chinensis DC. ActaPharmaceutica Sinica, 33(1): 37-41.6. Liu, P., and Yang, F. Y. (2002). Study on Anti-bacterial Endotoxin of Radix Bupleuri. Chinese Traditional Patent Medicine, 24(8): 527-528.7. Liu, Y. C., and Cong, P. C. (2006). Overview of Researches on Chemical Constituents and Pharmacological Effects of Bupleurum chinensisDC. Heilongjiang Medicine Journal, 19(3): 216-218.8. Luo, H. S., Zhao, Y. Y., and Qiao, L. (1996). Structural Identification of Saikosaponin P. Bulletin of Botany, 38 (11): 910-913.9. Shao, S. L. (2002). Preventive Effect of Radix bupleur and Rhizoma Curcumae longa on experimental Hyperlipidemia in Mice. Acta ChineseMedicine and Pharmacology, 30(4): 59-59.10. Thase, M. E. (2009). Treating Major Depression: Antidepressant Algorithms. Journal of Clinical Psychiatry, 70(12): 46-46.11. Wang, S. C., and Wang, L. (2002). Effects of Chaihu and Wulingwan on Chronic Liver Lesion in Mice. Journal of the Fourth MilitaryMedical University, 23(2): 133-136.12. Zhang, Y. (2000). Observation of Curative Effect of Bupleurum, ribavirin Injection in the Treatment of 56 Cases of Mumps. Tianjin Journal ofTraditional Chinese Medicine, 17(3): 45-45.13. Zheng, L. F., and Ming, L. (2005). The Study and Application of Animal Models of Depression. Anhui Medical and Pharmaceutical Journal,9(11): 801-803.14. Zhu, J. Q., Han, X. H., and Xue, Y. J. (2005). Effects of the Extracts from BCDC on Cellular VCR Concentration of Human HepatomaBEL-7402 Cells. Journal of Beihua University (Natural Science), 6(1): 62-64.

English

Pharmacological Experimental Study Of The Anti-Depressant Effect Of Total Saikosaponins.

ATHMSI Journals

PHARMACOLOGICAL EXPERIMENTAL STUDY OF THE ANTI-DEPRESSANT EFFECT OF TOTAL SAIKOSAPONINS.

Chai Hu has the hepato-protective, choleretic, anti-tussive, analgesic, anti-inflammatory, anti-viral, hypotensive, hypolipidemic, and anti-tumor pharmacological effects. In this study, the objective of this paper was to study the anti-depressant effect of total saikosaponins.Total saikosaponins were extracted by reflux method, and were identified by thin layer chromatography (TLC). The anti-depressant effect of total saikosaponins was investigated in vitro by tail suspension test, forced swimming test, and reserpine antagonism test in mice.Two times of reflux extraction, temperature of 70°C, and extraction time of 4hrs, for each extraction could improve the yield of saikosaponins. Each treatment group (100, 200, and 300 mg/kg), could significantly shorten the immobility time of mice in the tail suspension test in a somewhat dose-dependent manner. The total saikosaponins antagonized the reserpine-induced akinesia, and ptosis in mice.Total saikosaponins have an anti-depressant effect.SCI(E)PubMed0ARTICLEpingkew32@126.com2280-41

Liu Yu

Cao Chunmei

Ding Haijun

Pharmacological experimental study of the anti-depressant effect of total saikosaponins

https://journals.athmsi.org/index.php/ajtcam/article/download/2267/1819

PHARMACOLOGICAL EXPERIMENTAL STUDY OF THE ANTI-DEPRESSANT EFFECT OF TOTAL SAIKOSAPONINS.

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Institutional Repository of Peking University