Identification of Bupleurum marginatum var. Wall. ex DC stenophyllum (Wolff) Shan et Li and its analogues based on ITS2 barcode / 中草药

Objective: DNA barcoding technology, a molecular identification method, is applied to distinguish Bupleurum marginatum var. stenophyllum from its analogues in order to ensure the quality and clinical curative effect. Methods: In this study, the internal transcribed spacer 2 (ITS2) regions of 50 samples were amplified by PCR and sequenced bi-directionally. Obtained sequences were assembled using CodonCode Aligner. The genetic distances were computed by MEGA 6.0 in accordance with the kimura 2-parameter (K2P) model and the phylogenetic tree was constructed by Neighbor-joining (NJ) method. Moreover, the secondary structure of ITS2 was predicted using ITS2 database websites. Results: The intra-specific genetic distances were smaller than inter-specific ones in ITS2 regions of B. marginatum var. stenophyllum. The NJ tree and secondary structure results could distinctly differentiate B. marginatum var. stenophyllum and its analogues. Conclusion: ITS2 sequence can scientifically and reliably identify the authenticity of B. marginatum var. stenophyllum and could provide more new and reliable techniques to ensure clinical safety of this traditional Chinese medicine.

Triterpenoid saponins from Bupleurum marginatum var. stenophyllum / 中国中药杂志

Twelve compounds were obtained by phytochemical investigation of 70% EtOH ( containing 0.5%NH3•H2O )extract of the roots of Bupleurum marginatum var. stenophyllum. Based on comparison of their spectral data, including HR-ESI-MS, ¹H-NMR, ¹³C-NMR data, with those of the literature, their structures were elucidated as saikosaponin b2 (1), saikosaponin a(2), saikosaponin b1(3), saikosaponin d (4), hydroxysaikosaponin a (5), saikosaponin b3 (6), saikosaponin c(7),saikosaponin i (8), saikosaponin f (9), chikusaikosides Ⅱ(10), saikosaponin s (11), and saikosaponin I(12). All compounds belong to olean-type triterpenoid saponin and compounds 1, 3, 5, 8-9,11, and 12 were isolated from this plant for the first time. At a concentration of 20 μmol•L⁻¹, compounds 2, 4, 6, 8, 11 and 12 showed strong inhibition activity against influenza virus WSN33 with the inhibition rate of 91.3%,88.6%,53.4%,61.3%,77.3% and 57.4%,respectively.

Study on UPLC-MS Fingerprint of Bupleurum Marginatum in Northwest Hubei / 中国药师

Objective: To study the UPLC-MS fingerprint of Bupleurum marginatum DC. in northwest Hubei and establish the quality evaluation system for the herb. Methods:A UPLC-MS method was used to analyze 10 samples of B. marginatum DC with the following chromatographic conditions:an ACQUITY UPLC? RBEH C18 column (100 mm × 2. 1 mm, 1. 7 μm) was used, the mobile phase consisted of acetonitrile-0. 3% formic acid water with gradient elution, the flow rate was 0. 2 ml·min-1, and the detection wavelength was 203nm with ESI(-). Results:There were 8 common peaks in the UPLC-MS fingerprint of B. marginatum DC. Through analyzing the information of mass spectrometry and combining the references, 6 peaks were identified. Conclusion:The UPLC-MS fin-gerprint method is simple,rapid and feasible. The acquired UPLC-MS fingerprint of B. marginatum DC. and the evaluation indices can provide the scientific quality assessment of B. marginatum DC.

Comparison of Anti-inflammatory and Hepatoprotective Activities of Bupleurum marginatum Wall. ex DC. and Bupleurum chinense DC / 医药导报

Objective To compare anti-inflammatory and hepatoprotective effects of Bupleurum marginatum Wall. ex DC. and Bupleurum chinense DC. Methods The anti-inflammatory effect of two kinds of radix bupleuri decoction were compared using the xylene-induced ear edema mice model and the egg white-induced foot swelling rats model. Acute liver injury in mice was induced through intraperitoneal injection of peanut oil with 0. 1%carbon tetrachloride. Alanine aminotransferase ( ALT ) , aspartate aminotransferase ( AST ) , and alkaline phosphatase ( ALP ) in serum, superoxide dismutase ( SOD ) , malondialdehyde ( MDA ) , and glutathione ( GSH ) in liver tissue homogenate were detected. Histopathological changes, transforming growth factor-β ( TGF-β) and nuclear factor-κB ( NF-κB) in hepatic tissues were also observed. Results Ear swelling experiment showed that the decoction of both Bupleurum marginatum Wall. ex DC. and Bupleurum chinense DC. had equal anti-inflammatory effects (P>0. 05). Similarly,the decoction of Bupleurum marginatum Wall. ex DC. and Bupleurum chinense DC. improved levels of ALT,AST,SOD,GSH,and MDA to the same extent compared with the model control group ( P 0. 05 ). Histopathological and immunohistochemical experiments showed that expressions of TGF-β and NF- κB were similar between the two kinds of radix bupleuri. Conclusion Bupleurum marginatum Wall. ex DC. and Bupleurum chinense DC. have similar anti-inflammatory and hepatoprotective effects.

Content Variation of Bupleurum Polysaccharides from Bupleurum marginatum at Different Collecting Periods in Northwest of Hubei Province / 医药导报

Objective To study the dynamic change of polysaccharides from Bupleurum marginatum in Northwest of Hubei province,and to explore the optimal harvest period for Bupleurum marginatum. Methods The contents of soluble and crude Bupleurum polysaccharides were determined by anthrone-sulphuric acid colorimetric method and detected at 620 nm,and total polysaccharide content was calculated. Results The content of polysaccharides at various collecting periods was very different from 9. 642% to 15. 479%,and reached the highest level in November when the seed comes to mature. ConclusionThe optimal harvest period for Bupleurum marginatum in Northwest of Hubei is at the end of November.