Saikosaponin v-2(1). was isolated li om the roots of the title plant and thc structure was identified on rhs basis of spectral anal? sis. Saikosaponin v-2 is a new compound. which was identified as 3 beta .16 alpha .2...Saikosaponin v-2(1). was isolated li om the roots of the title plant and thc structure was identified on rhs basis of spectral anal? sis. Saikosaponin v-2 is a new compound. which was identified as 3 beta .16 alpha .23.28-tetrahydroxy-olean-11.13(18)-dien-30-oic acid-3-O-beta -D-glucopyranosyl- (1 -->2)glucopyranosyl-(1 -->3)-beta -D-fucopyranosol-30-O-xylitol ester.展开更多
A new chromone glycoside, saikochromoside A(1), was isolated from the roots of Bupleurum chinense DC. Its structure was determined on the bases of chemical and spectral analyses.
As a traditional Chinese herbal medicine exhibiting analgesic,fever-reducing and anti-inflammatory effects,Radix Bupleuri（Chai-Hu） is commonly used for the treatment of influenza,which is derived from the dried root...As a traditional Chinese herbal medicine exhibiting analgesic,fever-reducing and anti-inflammatory effects,Radix Bupleuri（Chai-Hu） is commonly used for the treatment of influenza,which is derived from the dried roots of Bupleurum chinense DC.and Bupleurum scorzonerifolium Willd.Among of diverse chemical components,saikosaponins are the key active components of the herb medicine.In the present study,we established a method of high performance liquid chromatography（HPLC） coupled with evaporative light scattering detection（ELSD） for simultaneous determination of saikosaponin a,c and d in root,stem,leaf and flower of Bupleurum chinense（B chinense） collected from different areas of Shanxi Province,China.The results from 16 samples of root,stem,leaf and flower of B chinense demonstrated that the total contents of the three saikosaponins in the root of B chinense collected from Dongshan Taiyuan,Xishan,Tianlongshan and Pangquangou were 4.26 mg/g,3.22 mg/g,4.23 mg/g and 3.05 mg/g,respectively.However,there was scarcely any saikosaponins in the stem,leaf and flower of B chinense collected from above-mentioned areas.The method of HPLC coupled with ELSD was suitable for quality control of Radix Bupleuri.The result also confirmed that the root of B chinense was the best medicinal part.展开更多
Objective: We are trying to verify how a transcription factor BcbZIP134 regulates the synthesis of saikosaponin using specific antibody. However, it is hard to obtain this soluble protein expressed in vitro, a prerequ...Objective: We are trying to verify how a transcription factor BcbZIP134 regulates the synthesis of saikosaponin using specific antibody. However, it is hard to obtain this soluble protein expressed in vitro, a prerequisite for antibody preparation. So we explored a condition in which the soluble protein can efficiently express followed by preparation of the specific polyclonal antibody.Methods: Firstly, the cDNA of BcbZIP134 from Bupleurum chinense was expressed in Transetta(DE3) E.coli. Different concentrations of IPTG(0.05 and 0.5 mmol/L) and different culture temperatures(16 and37 °C) were explored for efficient expression of target protein. Then, the expressed protein with His Tag was purified using Ni Sepharose 6 Fast Flow. Different concentrations of imidazole elution(15, 60, and300 mmol/L) were used to elute the target protein. The purified protein of BcbZIP134 was used to immunize rabbits. Using the purified polyclonal antibody, the expression of BcbZIP134 in transgenic B. chinense hairy root lines was analyzed by Western blot assays.Results: Under the conditions, IPTG 0.5 mmol/L, 16 °C, and overnight, recombinant protein of BcbZIP134 was obtained in high content using pEASY?-Blunt E1 vector and Transetta(DE3) E. coli. Anti-serum against BcbZIP134 was obtained with a high titer of 1: 51 200 after immunization in rabbits. The polyclonal antibody could react with BcbZIP134 overexpressed in transgenic B. chinense hairy root lines.Conclusion: An efficient protein expression system in E. coli was constructed and the soluble recombinant protein for BcbZIP134 was obtained. By using this protein, the specific and high-performance polyclonal antibody was prepared. Furthermore, by using BcbZIP134 overexpressed hairy roots, the availability of the polyclonal antibody was verified by Western Blotting.展开更多
基金This study was financially supported by the National Natural Science Foundation of China (29632050).
文摘Saikosaponin v-2(1). was isolated li om the roots of the title plant and thc structure was identified on rhs basis of spectral anal? sis. Saikosaponin v-2 is a new compound. which was identified as 3 beta .16 alpha .23.28-tetrahydroxy-olean-11.13(18)-dien-30-oic acid-3-O-beta -D-glucopyranosyl- (1 -->2)glucopyranosyl-(1 -->3)-beta -D-fucopyranosol-30-O-xylitol ester.
文摘A new chromone glycoside, saikochromoside A(1), was isolated from the roots of Bupleurum chinense DC. Its structure was determined on the bases of chemical and spectral analyses.
基金Shanxi Educational Committee(Grant No.20111113)Shanxi Science and Technology Department(Grant No.2016ZD0201)
文摘As a traditional Chinese herbal medicine exhibiting analgesic,fever-reducing and anti-inflammatory effects,Radix Bupleuri（Chai-Hu） is commonly used for the treatment of influenza,which is derived from the dried roots of Bupleurum chinense DC.and Bupleurum scorzonerifolium Willd.Among of diverse chemical components,saikosaponins are the key active components of the herb medicine.In the present study,we established a method of high performance liquid chromatography（HPLC） coupled with evaporative light scattering detection（ELSD） for simultaneous determination of saikosaponin a,c and d in root,stem,leaf and flower of Bupleurum chinense（B chinense） collected from different areas of Shanxi Province,China.The results from 16 samples of root,stem,leaf and flower of B chinense demonstrated that the total contents of the three saikosaponins in the root of B chinense collected from Dongshan Taiyuan,Xishan,Tianlongshan and Pangquangou were 4.26 mg/g,3.22 mg/g,4.23 mg/g and 3.05 mg/g,respectively.However,there was scarcely any saikosaponins in the stem,leaf and flower of B chinense collected from above-mentioned areas.The method of HPLC coupled with ELSD was suitable for quality control of Radix Bupleuri.The result also confirmed that the root of B chinense was the best medicinal part.
基金supported by the CAMS Innovation Fund for Medical Sciences(CIFMS)(2016-I2M-2-003)National Standardization Project of Traditional Chinese Medicine(ZYBZH-Y-JIN-34)
文摘Objective: We are trying to verify how a transcription factor BcbZIP134 regulates the synthesis of saikosaponin using specific antibody. However, it is hard to obtain this soluble protein expressed in vitro, a prerequisite for antibody preparation. So we explored a condition in which the soluble protein can efficiently express followed by preparation of the specific polyclonal antibody.Methods: Firstly, the cDNA of BcbZIP134 from Bupleurum chinense was expressed in Transetta(DE3) E.coli. Different concentrations of IPTG(0.05 and 0.5 mmol/L) and different culture temperatures(16 and37 °C) were explored for efficient expression of target protein. Then, the expressed protein with His Tag was purified using Ni Sepharose 6 Fast Flow. Different concentrations of imidazole elution(15, 60, and300 mmol/L) were used to elute the target protein. The purified protein of BcbZIP134 was used to immunize rabbits. Using the purified polyclonal antibody, the expression of BcbZIP134 in transgenic B. chinense hairy root lines was analyzed by Western blot assays.Results: Under the conditions, IPTG 0.5 mmol/L, 16 °C, and overnight, recombinant protein of BcbZIP134 was obtained in high content using pEASY?-Blunt E1 vector and Transetta(DE3) E. coli. Anti-serum against BcbZIP134 was obtained with a high titer of 1: 51 200 after immunization in rabbits. The polyclonal antibody could react with BcbZIP134 overexpressed in transgenic B. chinense hairy root lines.Conclusion: An efficient protein expression system in E. coli was constructed and the soluble recombinant protein for BcbZIP134 was obtained. By using this protein, the specific and high-performance polyclonal antibody was prepared. Furthermore, by using BcbZIP134 overexpressed hairy roots, the availability of the polyclonal antibody was verified by Western Blotting.
