The Yin and Yang actions of North American ginseng root in modulating the immune function of macrophages

<p>Abstract</p> <p>Background</p> <p>Immuno-modulatory effects of ginseng, including both immuno-stimulatory and immuno-suppressive effects, have been widely reported. This study aims to determine whether the paradoxical immuno-modulatory effect is related to unique phytochemical profiles of different North American (NA) ginseng, namely aqueous (AQ) and alcoholic (ALC) extracts.</p> <p>Methods</p> <p>AQ and ALC extracts were prepared and their immuno-bioactivity were studied <it>in vitro </it>in murine macrophages (Raw 264.7) through measuring the direct stimulatory production of pro-inflammatory mediator and cytokines as well as the suppression of lipopolysaccharide (LPS)-stimulatory response by the two extracts. Gel permeation chromatography was used to fractionate and isolate phytochemicals for characterization of ginseng extracts.</p> <p>Results</p> <p>AQ extract up-regulated the production of nitric oxide (NO), tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) while ALC extract did not. ALC extract but not AQ extract suppressed LPS-induced macrophage NO and TNF-α production. These immuno-stimulatory and suppressive effects were exhibited at similar extract concentrations. Moreover, the macrophage-stimulating activity of the AQ extract was inhibited in the presence of ALC extract. Fractionation of AQ extract revealed the presence of two major peaks at 230 nm with average molecular weights of 73,000 and 37,000 Da. The first fraction had similar elution volume as the crude polysaccharide (PS) fraction isolated from the AQ extract, and it was the only bioactive species. Parallel fractionation study of ALC extract yielded similar elution profiles; however, both sub-fractions were devoid of PS. Fraction I of the ALC extract suppressed LPS-induced NO production dose-dependently.</p> <p>Conclusion</p> <p>ALC extract of NA ginseng, which was devoid of PS, was immuno-inhibitory whereas the AQ extract, which contained PS, was immuno-stimulatory. These extract-related anti-inflammatory and pro-inflammatory effects may be considered as the Yin and Yang actions of ginseng.</p

Comparative pharmacokinetics and safety assessment of transdermal berberine and dihydroberberine

<div><p>The natural alkaloid berberine has been ascribed numerous health benefits including lipid and cholesterol reduction and improved insulin sensitivity in diabetics. However, oral (PO) administration of berberine is hindered by poor bioavailability and increasing dose often elicits gastro-intestinal side effects. To overcome the caveats associated with oral berberine, we developed transdermal (TD) formulations of berberine (BBR) and the berberine precursor dihydroberberine (DHB). These formulations were compared to oral BBR using pharmacokinetics, metabolism, and general safety studies <i>in vivo</i>. To complete this work, a sensitive quantitative LC-MS/MS method was developed and validated according the FDA guidelines for bioanalytical methods to simultaneously measure berberine, simvastatin, and simvastatin hydroxy acid with relative quantification used for the berberine metabolite demethylene berberine glucuronide (DBG). Acute pharmacokinetics in Sprague-Dawley rats demonstrated a statistically relevant ranking for berberine bioavailability based upon AUC_0-8 as DHB TD > BBR TD >> BBR PO with similar ranking for the metabolite DBG, indicating that transdermal administration achieves BBR levels well above oral administration. Similarly, chronic administration (14 days) resulted in significantly higher levels of circulating BBR and DBG in DHB TD treated animals. Chronically treated rats were given a single dose of simvastatin with no observed change in the drugs bioavailability compared with control, suggesting the increased presence of BBR had no effect on simvastatin metabolism. This observation was further supported by consistent CYP3A4 expression across all treatment groups. Moreover, no changes in kidney and liver biomarkers, including alanine aminotransferase and alkaline phosphatase, were observed between treatment formats, and confirming previous reports that BBR has no effect on HMG-CoA expression. This study supports the safe use of transdermal compositions that improve on the poor bioavailability of oral berberine and have the potential to be more efficacious in the treatment of dyslipidemia or hypercholesterolemia.</p></div

Extraction recovery and matrix effect of BBR, SIM and SHA (n = 6) and d6-BBR, d6-SIM and d6-SHA (n = 6x3).

<p>Extraction recovery and matrix effect of BBR, SIM and SHA (n = 6) and d6-BBR, d6-SIM and d6-SHA (n = 6x3).</p

LC-MS/MS parameter table.

<p>LC-MS/MS parameter table.</p

Clinical chemistry of liver and kidney function in chronically administered animals.

<p>Male Sprague-Dawley rats (N = 4/group) received once daily for 14 days administration of 90 mg/kg active via BBR oral gavage (BBR PO), 5% (w/w) BBR transdermal formulation (BBR TD), and 5% (w/w) DHB transdermal formulation (DHB TD) or vehicle control (Vehicle TD). Whole blood was collected on day 14 and analyzed for biomarkers of hepatic and nephrotic function including (A) ALT, (B) ALP, (C) creatinine, and (D) BUN. Error bars represent standard error of the mean.</p

Comparative acute pharmacokinetics of formulations.

<p>Male Sprague-Dawley rats received a single administration of 90 mg/kg of active via BBR oral gavage (BBR PO; ▲; N = 3), 5% (w/w) BBR transdermal formulation (BBR TD; -■-; N = 8), or 5% (w/w) DHB transdermal formulation (DHB TD; -●-; N = 8) and serum was collected over the course of eight hours. Error bars represent standard error of the mean.</p

LC-MS/MS profiles of serum samples with SIM and SHA introduced <i>in silico</i> and <i>in vitro</i>.

<p>Chromatograms of (A) blank serum (B) blank serum spiked with (1) d6-SHA (156.25 ng/mL), (2) SHA (62.5 ng/mL), (3) d6-SIM (156.25 ng/mL) and (4) SIM (62.5 ng/mL) and (C) serum samples 2.5 hours after oral administration of SIM (1) d6-SHA (spiked—156.25 ng/mL) (2) SHA (3) d6-SIM (spiked—156.25 ng/mL) and (4) SIM.</p

Comparative pharmacokinetics of formulations after 16 days of administration.

<p>Male Sprague-Dawley rats (N = 4/group) received once daily for 16 days administration of 90 mg/kg active via BBR oral gavage (BBR PO; ▼), 5% (w/w) BBR transdermal formulation (BBR TD; -■-), 5% (w/w) DHB transdermal formulation (DHB TD; -●-), or vehicle control (Vehicle TD; ▲) and serum collected over the course of nine hours. Error bars represent standard error of the mean.</p

Comparative chronic pharmacokinetic parameters of treatment groups.

<p>Comparative chronic pharmacokinetic parameters of treatment groups.</p