| In this paper, relevant reports of a material basis of Epimedium on antiosteoporosis efficacy were reviewed comprehensively, and then a variety of active flavonoids of Epimedium on anti-osteoporotic effects and potency were summarized. According to the researchs of our laboratory and the in vivo metabolism of activie flavonoids, the potential basis of Epimedium on anti-osteoporosis were aglycones and secondary glycosides. Meanwhile, the aglycones had a stronger anti-osteoporotic activity than its multi-glycosides. Suet oil (SO) was used to process Epimedium in traditional processing technology, which enhanced the role of Epimedium on kidney yang, and served to strengthen its anti-osteoporosis effect. Therefore, the synergistic anti-osteoporosis mechanisms of SO of aglycones (CIT, IT) were investigated.1. SO had no anti-osteoporotic effects itself, but it was able to enhance the anti-osteoporosis action of Epimedium flavonoid aglyconesOsteoporosis model was established by removing the bilateral ovaries of female rats, and the body weight changes of rats were recorded weekly. After the last administration, rats serum were obtained by orbital, and serum hydroxyproline (HOP) and alkaline phosphatase (AKP), osteoprotegerin (OPG), osteocalcin (OCN), nuclear factor-κB by body activator ligand (RANKL), tartrate-resistant acid phosphatase 5b (TRACP-5b) were measured. Subsequently, the rats were killed by cervical, and their uteri, the tibia and femur were collected, respectively. The femur and uterine weight were recorded, and the right femur were used to detecte the bone mineral density (BMD),3D-μCT scan and western blot assays, sequentially. Left femur were used to assay related gene and protein expression by quantitative real-time (RT-PCR). The right tibia were used to investigate the biomechanical properties by three-point bending test. Left tibia were carried out of immunohistochemistry experiments to study the expression of related factors. Our study found that CIT and IT both had anti-osteoporosis action. After adding SO, the anti-osteoporosis role of CIT and IT1 could be enhance. However, SO did not reflect the effect of anti-osteoporosis. We speculated that the synergies of SO should be attributed to the promotion in absorb for CIT and IT, and played a synergistic effect eventually.2. A preliminary study of the absorption, distribution, metabolism and excretion process (ADME) of anti-osteoporosis flavonoid aglyconesIn order to further research and development of CIT and IT on anti-osteoporotic effects, high-resolution mass spectrometry was used to analyze the metabolites and metabolic pathways of CIT and IT in rats after oral administration. As a result,59 metabolites of CIT were identified out, including 23 in the plasma,13 in bile,17 in urine, and 6 in feces; Weanwhile,43 metabolites of IT were identified, including 12 in plasma,14 in bile, and only its prototype in urine,17 in feces. The hydroxyls in the structure of the flavone might increase their binding affinity for forming the conjugates, which affected drug metabolism speed in the body, and ultimately led to the differences in pharmacological effects. Unfortunately, three phenolic hydroxyls were in the structure of IT (C-3,5 and 7), which was one more than CIT (C-3 and 5). Because of their differences in chemical structure, CIT and IT also had different pharmacological effects and efficacy strength.3. The synergy of SO was that it could promote the absorption of Epimedium flavonoids aglyconesIf SO can promote the absorption of CIT and IT, then the relative bioavailability of CIT and IT should be significantly improved under the role of SO. Therefore, the plasma concentration of CIT and IT were monitored after oral administration in rats, and the concentration-time curve were draw, and their pharmacokinetic parameters were calculated and evaluated. After adding SO, the Cmax and AUC of CIT and IT were significantly increased than a single CIT, IT group without SO. Therefore, SO could increase the bioavailability of CIT and IT.4. Promoting absorption of SO could be attributed to its rich in fatty acids, which had the surfactant-like actionSO was a fatty oil mainly derived from Ovis aries linnaeus or Capra hircus linnaeus, which was rich in inorganic elements and fatty acids. These fatty acids had a long-chain aliphatic and surfactants. Therefore, we analyzed the fatty acids composition by using GC-EI-qMS technology and professional database. Finally,25 kinds of fatty acids were identified, and the method of simultaneously determination of 10 major fatty acids were established. In addition, we also adopted the ICP-MS to establish a method for the simultaneous determination of 14 kinds of inorganic elements in SO. This work described the chemical composition of SO, which was conducive to establish the quality standards of SO.5. Fatty acids in SO and bile salt components secreted by body could self-assemble into nanometer mixed micelles, and which acted as a carrier and promoted the absorption of the aglyconesFatty acid components in SO exist hydrophilic group and hydrophobic group, which have some surface activity and a surfactant-like effect. Meanwhile, the normal body can secrete bile contain lots of bile acid or bile salts which are hydrophilic and hydrophobic. When CIT or IT together with suet oil orally entered into the body, the fatty acids of SO and the bile salts can self-assemble to a mixed micelles as a carrier of CIT or IT, which can improve absorption of CIT or IT. This hypothesis was also confirmed by being simulated in vitro.In summary, SO could be able to enhance the anti-osteoporosis effect of flavonoid aglycones, which was due to the surface active agents like effects of a large number of fatty acid compositions in SO (Stearic acid, Palmitic acid, Oleic acid). Meanwhile, under the combined action of bile salts, SO and bile salts could self-assemble into micelles in the body as a carrier of flavonoid aglycones, which promoted the intestinal absorption and efficiency of flavonoid aglycones. |
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