Establishment and application of quality evaluation method for Xiaochaihu Granules based on calibrator samples / 中国中药杂志

With reference to the production process documented in Chinese Pharmacopoeia, this paper prepared the calibrator samples of Xiaochaihu Granules from multiple batches and established a method for fingerprint analysis and content determination that could be used to evaluate Xiaochaihu Granules available in market. Multiple batches of Chinese herbal pieces contained in Xiaochaihu Granules were collected for preparing the calibrator samples according to the process in Chinese Pharmacopoeia. Following the establishment of fingerprints for calibrator samples by UHPLC, the method for determining the contents of saikosaponin B2, saikosaponin B1, baicalin, wogonoside, baicalein, liquiritin, glycyrrhizin G2 and glycyrrhizic acid in Xiaochaihu Granules was established. The experimental results showed that the fingerprints of calibrator samples had 26 common peaks, covering the chemical compounds of main herbs Bupleuri Radix, Scutellariae Radix, Changii Radix, Glycyrrhizae Radix et Rhizoma, and Rhizoma Zingiberis Recens. The similarity of fingerprints for 47 batches of Xiaochaihu Granules from 31 companies with the calibrator sample fingerprint ranged from 0.74 to 0.99, indicating good applicability of the established fingerprint. The contents of main components baicalin, saikosaponin B2, and glycyrrhizic acid in Xiaochaihu Granules were within the ranges of 22.917-49.108 mg per bag(RSD 19%), 0.28-2.19 mg per bag(RSD 62%), and 0.897-6.541 mg per bag(RSD 41%), respectively. The quality difference in saikosaponin B2, and glycyrrhizic acid among different manufacturers was significant. The fingerprint analysis and content determination method for calibrator samples of Xiaochaihu Granules prepared according to the production process in Chinese Pharmacopoeia has been proved suitable for evaluating the quality of Xiaochaihu Granules from different manufacturers. Saikosaponin B2, glycyrrhizic acid, and liquiritin should be added as content control indicators for Xiaochaihu Granules, aiming to further improve the product quality.

Effects of saikosaponin b_2 on inflammation and energy metabolism in mice with acute liver injury induced by LPS/GalN / 中国中药杂志

To study the effects of saikosaponin b2( SS-b2) on inflammatory factors and energy metabolism against lipopolysaccharide/galactosamine( LPS/Gal N) induced acute liver injury in mice. Mice were randomly divided into normal group( equal amount of normal saline),model group( 100 g·kg~(-1) LPS and 400 mg·kg~(-1) Gal N),low,medium,high dose group of SS-b2( SS-b25,10,20 mg·kg~(-1)·d-1) and positive control group( dexamethasone,10 mg·kg~(-1)). All of the groups except for the normal group were treated with LPS/Gal N though intraperitoneally injection to establish the acute liver injury model. The organ indexes were calculated. The levels of serum transaminases( ALT and AST) and the activities of ATPase( Na+-K+-ATPase,Ca2+-Mg2+-ATPase) in liver were detected. The activity of tumor necrosis factor-α( TNF-α),interleukin-1β( IL-1β) and interleukin-6( IL-6) were determined by the enzyme-linked immunosorbent assay( ELISA). The contents of lactate dehydrogenase( LDH) in liver were determined by micro-enzyme method. HE staining was used to observe the histopathological changes of the liver. Histochemical method was used to investigate the protein expression of liver lactate dehydrogenase-A( LDH-A). The protein expressions of Sirt-6 and NF-κB in the liver were detected by Western blot. According to the results,compared with the model group,there were significant changes in organ indexes in the high-dose group of SS-b2( P<0. 05). The level of ALT,AST,TNF-α,IL-1β,IL-6 and the activities of LDH in serum of mice with liver injury were significantly reduced in the medium and high dose groups of SS-b2( P<0. 01). With the increase of the concentration of SS-b2,the range of hepatic lesions and the damage in mice decreased. The activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase in liver of mice were significantly enhanced in each dose group( P<0. 01). The expression of NF-κB in liver tissues was significantly down-regulated in the medium and high dose group( P<0. 01). Meanwhile,the expression of Sirt-6 protein in the liver of mice with acute liver injury was significantly increased in each dose group( P<0. 01).In summary,SS-b2 has a significant protective effect on LPS/Gal N-induced acute liver injury in mice,which may be related to the down-regulation of NF-κB protein expression and up-regulation of Sirt-6 protein expression to improve inflammatory injury and energy metabolism.

Saikosaponin-b2 alleviates CCL4-induced acute liver injury in mice by inhibiting endoplasmic reticulum stress signal pathway / 中国药理学与毒理学杂志

OBJECTIVE To investigate the protective effect of saikosaponin-b2 (SS-b2) on acute liver injury induced by carbon tetrachloride (CCI4) and its inhibition on endoplasmic reticulum stress pathway in mice. METHODS Sixty male BALB/c mice were randomly divided into normal control, model, SS-b2 (5, 10 and 20 mg·kg-1) and silymarin (10 mg·kg-1) groups. Mice were ig given different drugs or saline for 7 d. Two hours after the last administration, mice were IP given 5% CCI4 to induce acute liver injury, except those in normal control group. The activities of glutamic-pyruvic transaminase (GPT), glutamic-oxaloacetic transaminase (GOT) and superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in mouse serum were detected by colorimetric method. Pathological changes in liver tissue were detected by HE staining. The expression levels of protein kinase R-like ER kinase (PERK), phospho-eukaryotic initiation factory 2a (p-elF2α), activating transcrIPtion factor-4(ATF4), glucose regulated protein 78 (GRP78) and CCAAT/enhancer-binding protein homologous protein (CHOP) were detected by immuno-histochemistry and Western blotting. RESULTS Compared with the normal control group, the activities of GPT and GOT and the content of MDA in mouse serum in model group were significantly increased (P<0.05, P<0.01), but the activity of SOD was significantly decreased (P<0.01). Meanwhile, the expression levels of PERK, p-elF2α, ATF4, GRP78 and CHOP in the liver tissue were significantly increased in the model group (P<0.01). Compared with the model group, SS-b2 groups significantly reduced the activity of GPT, GOT and the content of MDA in the serum of acute liver injury mice (P<0.05, P<0.01), but increased the activity of SOD (P<0.01), and the expression levels of endoplasmic reticulum stress pathway proteins mentioned above were significantly reduced (P<0.05, P<0.01). HE staining results showed that different concentrations of SS-b2 could significantly improve the swelling of hepatocytes, the dissolution of nuclei around hepatic lobules, and the arrangement disorder of hepatic cords induced by CCI4 in mice. The cell morphology was obviously improved. CONCLUSION SS-b2 has a significant protective effect on CCL-induced acute liver injury in mice. The mechanism may be related to the inhibition of oxidative stress and down-requlation of the expressions of ERS related protein.

Study on saponins from Reineckia carnea belongs to Guizhou Miao medicine / 中草药

Objective: To study the saponins from Reineckia carnea of Guizhou Miao medicine. Methods: The chemical constituents from 95% EtOH extract were isolated and purified by repeated silica gel column chromatography, macroporous resin, ODS C18, Sephadex LH-20 gel column chromatography, and Pre-HPLC, and their structures were identified on the basis of physical and chemical properties, HR-ESI-MS and NMR spectral analysis. Results: Ten compounds were isolated and identified as (1β,3β,16β,22S)-cholest-5-ene-1,3,16,22-tetrol1,16-di (β-D-glucopyranosede) (1), (1β,3β,16β,22S)-cholest-5-ene-1,3,16,22-tetrol-1-[O-α-L-rhamno-pyranosyl-(1→2)-β-D-glucopyranoside]-16-(β-D-glucopyranoside) (2), (20S,22R)-spirost-25(27)-en-1β,3β,4β,5β-tetraol-5-O-β-D-glucopyranoside (3), kitigenin 5-O-β-D-glucopyrannoside (4), aspidistrin A (5), (17,20-S-trans)-5β-pregn-16-en-1β,3β-diol-20-one-1-O-β-D-xylopyranosyl-(1→2)-[α-L-rhamnopyranosyl]-3-O-α-L-rhamnopyranoside (6), diosgenin-3-O-β-D-glucopyranoside (7), ophiopogonin T (8), prosaikogenin D (9), and saikosaponin b2 (10). Conclusion: Compounds 3, 5, 8, 9, and 10 are isolated from this plant for the first time, which provided some reference for further elucidation and development of saponins from R. carnea.

Simultaneous determination of four saikosaponins in Xiaozheng Pellets by QAMS / 中成药

AIM To establish a quantitative analysis of multi-components by single-marker (QAMS) method for the simultaneous content determination of four saikosaponins in Xiaozheng Pellets (Bupleuri Radix,Cyperi Rhi-zoma,Rhei Radix et Rhizoma,etc.).METHODS The analysis of 5％ ammonia-methanol extract of this drug was performed on a Waters Xbridge C18column (250 mm ×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-water flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 210 nm and 254 nm.With saikosaponin a as a internal standard,the relative correction factors of saikosaponins b1,b2 and c were calculated,followed by the determination of their contents.RESULTS Four saikosaponins showed good linear relationships within their own ranges (r ≥ 0.999 5),whose average recoveries were 98.08％-102.94％ with the RSDs of 0.85％-1.82％.The results obtained by QAMS approximated those obtained by external standard method.CONCLUSION This simple,precise and feasible method can be used for the quality control of Xiaozheng Pellets.