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Purification of saponin compounds in Bupleurum falcatum by solvent partitioning and preparative LC.

Saponin compounds (saikosaponin c, a, and d) in Bupleurum falcatum were partially purified by solvent partitioning of the herbal extract using diethyl ether, distilled water, n-butanol, and acetone. After separation of the saponins by preparative LC, the purity of each saikosaponin was more than 94%. The identities of purified individual saikosaponins were confirmed by TLC, analytical LC, and fast-atom bombardment mass spectrometry.

Bupleurum↗

Effects of a Quillaja saponaria extract on growth performance and immune function of weanling pigs challenged with Salmonella typhimurium.

Ninety-six pigs (initially 8.9 kg and 24 d of age) were used in a 28-d experiment to determine the effects of Quillaja saponaria extract (QS) on weanling pig growth performance and immune function in response to enteric disease challenge with Salmonella typhimurium (ST). Experimental treatments were arranged in a 2 x 4 factorial with main effects of disease challenge (control vs ST-challenge) and dietary addition of QS (0, 125, 250, or 500 mg/kg). Pigs were fed QS diets for 14 d and then challenged orally with ST or sterile media. There were no differences in ADG or ADFI among dietary treatments, but gain/feed ratio (G/ F) was depressed (P < 0.06) in pigs fed 250 mg/kg QS. ST-challenge reduced ADG (P < 0.05), ADFI (P < 0.05), and G/F (P < 0.05) 1 wk after challenge. Daily estimates revealed reductions in feed intake in ST-infected pigs on d 2 to 5 following infection (P < 0.05), and rectal temperature was increased maximally 2 d following infection (P < 0.05). There was a marked decline in serum IGF-I during the 6 d after ST-infection (P < 0.05). ST-challenge produced a rise (P < 0.05) in serum haptoglobin on d 7 after challenge, and serum alpha1-acid glycoprotein (AGP) in ST-challenged pigs also was elevated (P < 0.05) above controls on d 7 and 14 after challenge. Serum immunoglobulin (Ig) M increased (P < 0.05) over time in both groups, and serum IgM of ST-challenged pigs was greater than controls on d 7 after challenge (P < 0.05). Serum IgG was not affected by enteric disease challenge; however, on d 7 and 14 after disease challenge, serum IgG for both groups was greater (P < 0.05) than on d 0. Dietary QS had no significant influence on any of the end points used to characterize the acute phase response to ST-challenge. Phagocytic cell function was depressed in pigs fed 250 (P < 0.05) and 500 (P < 0.05) mg/kg as compared to pigs fed 125 mg/kg QS. Yet, there was no difference in phagocytic function among pigs fed 0, 250, or 500 mg/kg QS. We conclude that this model of enteric disease invokes an acute phase response accompanied by decreases in feed intake and serum IGF-I. Furthermore, dietary QS, at the levels fed in this study, appears to offer little benefit to growth performance or immune function in the presence or absence of ST-challenge.

Acute-Phase Proteins↗

Effect of feeding Sarsaponin in cattle and swine diets.

In four trials, steer calves were received in the feedlot, processed and fed diets supplemented with soybean meal (SBM), 1% urea (UR) or 1% urea plus sarsaponin (S) over a 28-d period. In trials 1 and 2, the feeding period was extended to approximately 62 d, in which steers were fed a common (SBM) diet the last 34 d. In trials 3 and 4, a SBM plus S diet treatment was included. During the first 28 d (four trial summary) daily gains of steers fed urea plus S (.74 kg) were intermediate to and significantly different from gains of steers fed SBM (.84 kg) or UR (.66 kg) diets. However, at the end of the 62-d feeding period (two trial summary) daily gains, feed intakes and feed efficiency did not differ (P greater than .05) among treatments. No significant improvements in performance were found in steers fed SBM diets supplemented with S. In swine trials, pigs were fed diets containing no additive, 63 mg S X kg-1, 55 mg chlortetracycline (C) X kg-1 or S plus C in a grower-finisher (GF) and grower (G) trial. In the GF trial, overall efficiency of feed conversion was improved (P less than .05) by feeding S or S plus C. In the G trial, daily gains and intakes were greatest for pigs fed S plus C and differed (P less than .1) from those of pigs fed S or C in the diet. Compared with feeding S or C alone, gain and intake of growing pigs were stimulated to a greater extent when S was fed in combination with C. Feeding S with or without C improved efficiency of feed conversion in finishing pigs.

Animal Feed↗

Fragility of erythrocytes in animals affected by lantana poisoning.

Clinical observations on four male cow calves and three other lantana poisoned animals under field conditions have been made along with hematological studies, including observations on plasma bilirubin content and osmotic fragility of erythrocytes. Four healthy male cow calves served as control. Five clinically advanced cases succumbed within 1 to 10 days of observation. Hematocrit values in four of the seven affected animals were abnormally high. The total plasma bilirubin content in the fatal cases was of the order of 5 to 50 times the normal value while, in calves that recovered, it was not more than 3 times. The osmotic fragility of erythrocytes was measured by the rate of hemolysis in decreasing concentrations of saline. The erythrocytes of normal animals resisted hemolysis till the saline concentration was lowered to 0.60%, and it was complete at the low saline concentration of 0.30%. However, in lantana poisoned animals, hemolysis started even in 0.72% saline and was complete at a saline concentration of 0.44%. The hemolysis curves of animals that died were far removed from the normal, but those of the calves that recovered and the one that died but had the lowest bilirubin content closely approached the normal curve.

Animals↗

Characteristics and mechanism of enzyme secretion and increase in [Ca2+]i in Saikosaponin(I) stimulated rat pancreatic acinar cells.

AIM: This investigation was to reveal the characteristics and mechanism of enzyme secretion and increase in [Ca2+]i stimulated by saikosaponin(I) (SA(I)) in rat pancreatic acini. METHODS: Pancreatic acini were prepared from male Wistar rats. Isolated acinar cells were suspended in Eagle's MEM solution. After adding drugs, the incubation was performed at 37 degrees for a set period of time. Amylase of supernatant was assayed using starch-iodide reaction. Isolated acinar single cell was incubated with Fura-2/AM at 37 degrees, then cells were washed and resuspended in fresh solution and attached to the chamber. Cytoplasm [Ca2+]i of a single cell was expressed by fluorescence ratio F340/F380 recorded in a Nikon PI Ca2+ measurement system. RESULTS: Rate course of amylase secretion stimulated by SA(I) in rat pancreatic acini appeared in bell-like shape. The peak amplitude increased depended on SA(I) concentration. The maximum rate responded to 1 x 10(-5)mol/L SA(I) was 13.1-fold of basal and the rate decreased to basal level at 30 min. CCK-8 receptor antagonist Bt(2)-cGMP markedly inhibited amylase secretion stimulated by SA(I) and the dose-effect relationship was similar to that by CCK-8. [Ca2+]i in a single acinar cell rose to the peak at 5 min after adding 5 x 10(-6)mol/L SA(I) and was 5.1-fold of basal level. In addition, there was a secondary increase after the initial peak. GDP could inhibit both the rate of amylase secretion and rising of [Ca2+]i stimulated by SA(I) in a single pancreatic acinar cell. CONCLUSION: SA(I) is highly efficient in promoting the secretion of enzymes synthesized in rat pancreatic acini and raising intracellular [Ca2+]i. Signaling transduction pathway of SA(I) involves activating special membrane receptor and increase in cytoplasm [Ca2+]i sequentially.

Amylases↗

Steroids and steroid metabolism in plant tissue cultures.

The steroids which have been isolated from plant callus and suspension cultures are reviewed. In addition, the research involving the use of plant tissue cultures to investigate steroid biosynthesis and metabolism is summarized.

Biotransformation↗

[Saponins in Bupleurum smithii Wolff var. parvifolium Shan et Y. Li].

Five saikosaponins and two saikosapogenins were isolated from the roots of Bupleurum smithii var. parvifolium by column chromatography, preparative-TLC and HPLC, and identified as saikosaponin a, saikosaponin d, saikosaponin b2, saikosaponin b4, chikusaikoside I, saikogenin F and saikogenin G on the basis of spectral analysis. All of them were isolated from this plant for the first time.

Bupleurum↗

[Saikosaponin-d up-regulates GR mRNA expression and induces apoptosis in HL-60 cells].

OBJECTIVE: To research the effect of saikosaponin-d (SSd) on the expression of glucocorticoid receptor (GR) mRNA and induction of apoptosis in HL-60 cells. METHODS: Antiproliferation effects of SSd on HL-60 cells were determined by 3H-thymidine incorporation. Apoptosis of the SSd-treated cells was determined by DNA fragmentation analysis and flow cytometry. GR mRNA was analyzed by Northern blot analysis. RESULTS: After 48 h treatment with 10 micrograms/ml SSd, 3H-thymidine incorporation was decreased in HL-60 cells and the effects were time and dose dependent. DNA ladder appeared after 60 h treatment with SSd. The flow cytometry analysis showed HL-60 cells was arrested at G0/G1 phase and apoptotic peak appeared in sub-G1 phase. The expression of GR mRNA was increased. CONCLUSION: SSd up-regulates the expression of GR mRNA, inhibits the cell growth and induces apoptosis in HL-60 cells.

Antineoplastic Agents, Phytogenic↗

[Effect of Saikosaponin-d on up-regulating GR mRNA expression and inhibiting cell growth in human leukemia cells].

OBJECTIVE: To study the effect of Saikosaponin-d (SSd) on the expression of glucocorticoid receptor (GR) mRNA and cell growth in HL60 cells. METHODS: Antiproliferation effects of SSd on HL60 cells were determined by 3H-thymidine incorporation. Cell cycle analysis was also performed by flow cytometry. GRmRNA was analyzed by northern blot analysis. RESULTS: After 48 hours treatment with 10 micrograms/ml SSd, 3H-thymidine incorporation decreased in HL60 cells and the effect was time and dose dependent. The flow cytometry analysis showed HL60 cells were arrested at G0/G1 phase. The expression of GR mRNA increased. CONCLUSION: SSd exhibits the inhibition effects on cell growth in HL60 cells and could up-regulate the expression of GR mRNA in HL60 cells.

Anti-Inflammatory Agents↗

[A new saikosaponin from Bupleurum Chinese DC].

Bupleurum chinese DC. is a well-known and very important traditional Chinese drug. It is often used to treat common cold with fever, alternating chill and fever, the feeling of fullness and oppression in the chest. A new saikosaponin was isolated from Bupleurum chinese DC. and its structure was identified as 3 beta, 16 beta, 28-trihydroxy-11-alpha-methoxy-olean-12-ene-3-O-beta-D-glucosyl-(1-->3)-beta D-fucoside on the basis of chemical and spectral evidence. In addition, six known saikosaponins were isolated and characterized as saikosaponin a, saikosaponin d, saikosaponin c, saikosaponin f, saikosaponin b3 and saikosaponin b2. All these compounds were isolated from Bupleurum chinese DC. for the first time.

Bupleurum↗

[A new saikosaponin from Bupleurum chinense DC].

Bupleurum chinense DC. is a well-known and very important traditional Chinese drug. It is often used to treat common cold with fever, alternating chill and fever, feeling of fullness and oppression in the chest. A new saikosaponin was isolated from Bupleurum chinense DC. and its structure was identified as 3-O-[beta-D-glucopyranosyl (1-->3)-beta-D-fucopyranosyl]-3 beta, 16 alpha, 23, 28-tetrahydroxy-olean-11, 13 (18)-dien-30-oic acid 30-O-[xylityl(1-->1)]-beta-D-glucopyranosyl-6-ester on the basis of chemical and spectral evidence. In addition, three known saikosaponins were isolated and characterized as saikosaponin 1, 2"-O-acetyl-saikosaponin b2 and 2"-O-acetyl-saikosaponin a. All the compounds were isolated from Bupleurum chinense DC. for the first time.

Bupleurum↗

Different contribution of apoptosis to the antiproliferative effects of diosgenin and other plant steroids, hecogenin and tigogenin, on human 1547 osteosarcoma cells.

Regulation of growth arrest and apoptosis are, in part, controlled by the tumor suppressor p53 after its phosphorylation which causes a determinant role in its functional activation. Moreover, PPAR regulate many functions such as proliferation and apoptosis. We compared the biological activity of diosgenin with hecogenin and tigogenin, plant steroids structurally close to diosgenin, on proliferation rate, cell cycle distribution and apoptosis in human 1547 osteosarcoma cells. We found that all three molecules have an antiproliferative effect but gel shift analysis demonstrated that none of the plant steroids transactivated PPAR in human 1547 osteosarcoma cells whereas these molecules induced NF-kappaB binding to DNA. Although these plant steroids have a very close structure, only diosgenin caused a cell cycle arrest associated with strong apoptosis. This biological action seems correlated with a large increase of p53 protein expression. This fact was showed by immunofluorescence analysis which confirmed that diosgenin strongly enhanced the activation of p53 in contrast to hecogenin and tigogenin actions.

Antineoplastic Agents↗

[The identification and determination of marker substances in xiaoyao wan by three dimensional HPLC].

A Chinese traditional patent medicine Xiaoyao Wan was analysed by three dimensional HPLC. The result shows that the Chinese traditional medicines involved in Xiaoyao Wan were identified excellently and the content of marker substances can be determined accurately. According to the results of determination, the content of saikosaponin b1, b2, glycyrrhizin, paeoniflorin and atractylenolide III in Xiaoyao Wan was 0.43, 0.53, 5.22, 3.95 and 0.16 (mg/g), respectively; The RSD was 1.2, 0.8, 0.4, 0.3 and 1.0 (%) respectively.

Benzoates↗

[A histochemical study of saikosaponins].

The root of Bupleurum chinese and B.scorzonerifolium was examined histochemically in order to clarify the distribution of saikosaponins in various tissues and parts of the root. By means of qualitative and quantitative analyses, saikosaponins, the bioactive principles in the root, were found to be abundant in the cortex outside the cambium and very little in the xylem of the root. It was also found that the thinner root hairs had the highest contents of saikosaponins, but decreased towards the thicker root head.

Anti-Inflammatory Agents, Non-Steroidal↗

Immunoenhancing activity of protopanaxatriol-type ginsenoside-F3 in murine spleen cells.

AIM: To investigate the immunoenhancing activity of ginsenoside-F3 in murine spleen cells and explore its mechanism. METHODS: The enhancing effect of ginsenoside-F3 on murine spleen cell proliferation was studied using [3H]thymidine incorporation assay. Effects of ginsenoside-F3 on the production of type 1 cytokines IL-2, IFN-gamma, and type 2 cytokines IL-4 and IL-10 from murine spleen cells were detected by ELISA method. Effects of ginsenoside-F3 on mRNA level of cytokines IL-4, IFN-gamma, and transcription factors T-bet and GATA-3 were evaluated by RT-PCR analysis. Effect of ginsenoside-F3 on NF-kappaB DNA binding activity in murine spleen cells was investigated by electrophoretic mobility shift assays (EMSA). RESULTS: Ginsenoside-F3 at 0.1-100 micromol/L not only promoted the murine spleen cell proliferation, but also increased the production of IL-2 and IFN-gamma, while decreased the production of IL-4 and IL-10 from murine spleen cells with the maximal effect at 10 micromol/L. RT-PCR analysis displayed that ginsenoside-F3 enhanced the IFN-gamma and T-bet gene expression and decreased IL-4 and GATA-3 gene expression. EMSA experiment showed that ginsenoside-F3 10 micromol/L enhanced the NF-kappaB DNA binding activity induced by ConA in murine spleen cells. CONCLUSION: Ginsenoside-F3 has immunoenhancing activity by regulating production and gene expression of type 1 cytokines and type 2 cytokines in murine spleen cells.

Animals↗