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Effect of nanopowdered oyster shell on osteoporosis in rats
Abstracts of Pharma Nutrition 2013 / PharmaNutrition 2 (2014) 75–119
were 8.1 and 6.8% lower, respectively, in NRG than that in PRG with significant differences (p < 0.05). IgM (5532.46 ng/ml) was 8.2% higher in serum of NRG than that in PRG with significant difference from that of the control, and IgG was also observed similar to IgM (p < 0.05). In conclusion, NRG treatment showed to have an improved preventive efficacy on CIA mice, therefore, NRG was considered it is worth functional ingredient for RA. Keywords: Nanopowdered red ginseng; Collagen-induced arthritis; Ginsenoside http://dx.doi.org/10.1016/j.phanu.2013.11.107 [P67] Effect of nanopowdered oyster shell on osteoporosis in rats Y.K. Lee, D.H. Kim, H.S. Kwak ∗ Sejong University, Republic of Korea Oyster shell is one of potential sources of dietary calcium that had been studied as an alternative treatment for osteoporosis. Osteoporosis is major health threat to the elderly women throughout the world by a decrease in bone mass and deterioration of bone tissue, resulting in bone fragility and susceptibility to fractures. Nanosized oyster shell can enhance potential effects and bioavailability. Therefore, this study was designed to the effect of nanopowdered oyster shell (NPOS) on ovariectomized osteoporosis rats. Thirty two 7-week old Sprague-Dawley rats were divided into six groups, sham, ovariectimized, fed diets containing NPOS, Zn-activated nanopowdered oyster shell (Zn-NPOS) and powdered oyster shell (POS). After 60 days of feeding, the concentration of calcium in serum (10 mg/dL), urine (12 mg/dL), liver (15 mg/dL), kidney (24 mg/dL), femur (13,612 mg/dL) and fecal (12751 mg/dL) from the NPOS and Zn-NPOS were significantly higher compared with POS. The osteocalcin ratio was 1.8 and 2.0 pg/mL higher fed in NPOS and Zn-NPOS rats, respectively. However, deoxypyridinoline (38pmole/L) and n-telopeptide of type collagen (3 nM BCE) were significantly decreased in NPOS and Zn-NPOS. Micro-CT analysis of femur confirmed that NPOS and Zn-NPOS groups exhibited greater bone mineral density (BMD) than that of the POS group. The structural parameters of trabecular bone for bone volume (BV/TV), bone surface (BS/BV), trabecular thickness (Tb.Th), trabecular number (Tb.N), and treabecular separation (Tb.Sp) in fed with NPOS and Zn-NPOS were significantly higher than the OVX, respectively. The present study indicated that NPOS treatment can be effective on enhance bone calcium metabolism for osteoporosis rats. Therefore, the NPOS and Zn-NPOS may be effective alternative treatment for osteoporosis. Keywords: Nanopowdered Oyster Shell, Osteoporosis in rats, Bone analysis http://dx.doi.org/10.1016/j.phanu.2013.11.108 [P68] Effects of nanopowdered eggshell on postmenopausal osteoporosis: A rat study M.A. Mijan, Y.K. Lee, D.H. Kim, H.S. Kwak ∗ Sejong University, Republic of Korea Eggshell has received a great deal of attention over the last few years for showing preventing effects against postmenopausal osteoporosis. These potential effects of eggshell can be further enhanced by reducing the size to nano level. Therefore, this study was carried out to investigate the preventive effects of nanopowdered eggshell (NPES) on postmenopausal osteoporo-
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sis in ovariectomized rats. Seven-week old female rats were ovariectomized and assigned to 4 groups: sham operated (sham), ovariectomized rats treated with vehicle (OVX), ovariectomized rats treated with NPES (5 g/kg body wt), and ovariectomized rats treated with powdered eggshell (PES) (5 g/kg body wt). The rats fed with NPES and PES exhibited 6.6% and 2.2% greater bone mineral densities (BMD) than the OVX rats. Femoral microstructure manifested that the NPES group had a remarkably greater trabecular area than the PES and OVX groups. Investigation of the trabecular bone in NPES ingested rats revealed a 12.4% higher bone volume (BV/TV), 7.5% higher trabecular number (Tb.N), and 14.5% lower trabecular separation (Tb.Sp) compared to OVX rats. Serum analysis demonstrated that NPES-fed rats showed a significantly higher (22.4%) osteocalcin level than the OVX rats. Urine analyses in NPESsupplemented rats revealed a 43.7% lower deoxypyridinoline (DPD) and 87% lower Nteleopeptide of type I collagen (NTX) than OVX rats. Collectively, these results indicated a high level of pharmaceutical effects in preventing ovariectomy induced severe bone loss in rats. Therefore, it is worth concluding that the present study has demonstrated that nanopowdered eggshell could be recognized as an effective alternative treatment for postmenopausal osteoporosis. Keywords: Nanopowdered eggshells; Postmenopausal osteoporosis; Ovariectomized rats http://dx.doi.org/10.1016/j.phanu.2013.11.109 [P69] Impact of quercetin on hepatic miRNA expression in rats S. Wein ∗ , M. Kluth, S. Wolffram University of Kiel, Germany The flavonol quercetin is well recognized for its multiple biological effects including influences on gene expression. Until now, however, no comprehensive studies on possible influences of quercetin on miRNA expression are available. Mature miRNAs interact with transcripts by binding to RNA and thus influences mRNA translation and/or degradation. We investigated effects of quercetin on hepatic miRNA in rats fed high-fat and low-fat diets. Male Wistar rats (n = 32) were kept for 7 weeks either on a highfat (22% fat, HF) or a low-fat diet (4% fat, LF) without or with the addition of 0.01% quercetin (HFQ, LFQ). Hepatic expression of 352 miRNA was measured using the RT2 miRNA PCR Array System (SABiosciences). All the samples passed the quality controls based on the Ct values of reverse transcription controls and 2 positive PCR controls. Any threshold cycle (Ct) value less than 35 was considered positive call. Relative miRNA expression levels were determined with the Ct method, normalized with the housekeeping genes Rnu6, rno-U87, which were geometrically averaged and reported as 2−Ct, where Ct = (target miRNA) − Ct (housekeeping). Expression was considered different, if fold change was >3. Different relative expression was found in 228 miRNA. Using the TargetScan database, we identified potential target genes of the miRNA influenced by quercetin. Thus we could link 35 miRNA to lipid metabolism, and 15 miRNA to carbohydrate metabolism. Two miRNA (rno-miR-19a, rno-miR-19b) have potential target genes involved in lipid as well as in carbohydrate metabolism. We conclude from the present study, that quercetin influences hepatic miRNA concentrations in a complex way and that parts of the multiple biological effects of quercetin might be mediated by this pathway. However, it is not possible at the moment to spec-
were 8.1 and 6.8% lower, respectively, in NRG than that in PRG with significant differences (p < 0.05). IgM (5532.46 ng/ml) was 8.2% higher in serum of NRG than that in PRG with significant difference from that of the control, and IgG was also observed similar to IgM (p < 0.05). In conclusion, NRG treatment showed to have an improved preventive efficacy on CIA mice, therefore, NRG was considered it is worth functional ingredient for RA. Keywords: Nanopowdered red ginseng; Collagen-induced arthritis; Ginsenoside http://dx.doi.org/10.1016/j.phanu.2013.11.107 [P67] Effect of nanopowdered oyster shell on osteoporosis in rats Y.K. Lee, D.H. Kim, H.S. Kwak ∗ Sejong University, Republic of Korea Oyster shell is one of potential sources of dietary calcium that had been studied as an alternative treatment for osteoporosis. Osteoporosis is major health threat to the elderly women throughout the world by a decrease in bone mass and deterioration of bone tissue, resulting in bone fragility and susceptibility to fractures. Nanosized oyster shell can enhance potential effects and bioavailability. Therefore, this study was designed to the effect of nanopowdered oyster shell (NPOS) on ovariectomized osteoporosis rats. Thirty two 7-week old Sprague-Dawley rats were divided into six groups, sham, ovariectimized, fed diets containing NPOS, Zn-activated nanopowdered oyster shell (Zn-NPOS) and powdered oyster shell (POS). After 60 days of feeding, the concentration of calcium in serum (10 mg/dL), urine (12 mg/dL), liver (15 mg/dL), kidney (24 mg/dL), femur (13,612 mg/dL) and fecal (12751 mg/dL) from the NPOS and Zn-NPOS were significantly higher compared with POS. The osteocalcin ratio was 1.8 and 2.0 pg/mL higher fed in NPOS and Zn-NPOS rats, respectively. However, deoxypyridinoline (38pmole/L) and n-telopeptide of type collagen (3 nM BCE) were significantly decreased in NPOS and Zn-NPOS. Micro-CT analysis of femur confirmed that NPOS and Zn-NPOS groups exhibited greater bone mineral density (BMD) than that of the POS group. The structural parameters of trabecular bone for bone volume (BV/TV), bone surface (BS/BV), trabecular thickness (Tb.Th), trabecular number (Tb.N), and treabecular separation (Tb.Sp) in fed with NPOS and Zn-NPOS were significantly higher than the OVX, respectively. The present study indicated that NPOS treatment can be effective on enhance bone calcium metabolism for osteoporosis rats. Therefore, the NPOS and Zn-NPOS may be effective alternative treatment for osteoporosis. Keywords: Nanopowdered Oyster Shell, Osteoporosis in rats, Bone analysis http://dx.doi.org/10.1016/j.phanu.2013.11.108 [P68] Effects of nanopowdered eggshell on postmenopausal osteoporosis: A rat study M.A. Mijan, Y.K. Lee, D.H. Kim, H.S. Kwak ∗ Sejong University, Republic of Korea Eggshell has received a great deal of attention over the last few years for showing preventing effects against postmenopausal osteoporosis. These potential effects of eggshell can be further enhanced by reducing the size to nano level. Therefore, this study was carried out to investigate the preventive effects of nanopowdered eggshell (NPES) on postmenopausal osteoporo-
113
sis in ovariectomized rats. Seven-week old female rats were ovariectomized and assigned to 4 groups: sham operated (sham), ovariectomized rats treated with vehicle (OVX), ovariectomized rats treated with NPES (5 g/kg body wt), and ovariectomized rats treated with powdered eggshell (PES) (5 g/kg body wt). The rats fed with NPES and PES exhibited 6.6% and 2.2% greater bone mineral densities (BMD) than the OVX rats. Femoral microstructure manifested that the NPES group had a remarkably greater trabecular area than the PES and OVX groups. Investigation of the trabecular bone in NPES ingested rats revealed a 12.4% higher bone volume (BV/TV), 7.5% higher trabecular number (Tb.N), and 14.5% lower trabecular separation (Tb.Sp) compared to OVX rats. Serum analysis demonstrated that NPES-fed rats showed a significantly higher (22.4%) osteocalcin level than the OVX rats. Urine analyses in NPESsupplemented rats revealed a 43.7% lower deoxypyridinoline (DPD) and 87% lower Nteleopeptide of type I collagen (NTX) than OVX rats. Collectively, these results indicated a high level of pharmaceutical effects in preventing ovariectomy induced severe bone loss in rats. Therefore, it is worth concluding that the present study has demonstrated that nanopowdered eggshell could be recognized as an effective alternative treatment for postmenopausal osteoporosis. Keywords: Nanopowdered eggshells; Postmenopausal osteoporosis; Ovariectomized rats http://dx.doi.org/10.1016/j.phanu.2013.11.109 [P69] Impact of quercetin on hepatic miRNA expression in rats S. Wein ∗ , M. Kluth, S. Wolffram University of Kiel, Germany The flavonol quercetin is well recognized for its multiple biological effects including influences on gene expression. Until now, however, no comprehensive studies on possible influences of quercetin on miRNA expression are available. Mature miRNAs interact with transcripts by binding to RNA and thus influences mRNA translation and/or degradation. We investigated effects of quercetin on hepatic miRNA in rats fed high-fat and low-fat diets. Male Wistar rats (n = 32) were kept for 7 weeks either on a highfat (22% fat, HF) or a low-fat diet (4% fat, LF) without or with the addition of 0.01% quercetin (HFQ, LFQ). Hepatic expression of 352 miRNA was measured using the RT2 miRNA PCR Array System (SABiosciences). All the samples passed the quality controls based on the Ct values of reverse transcription controls and 2 positive PCR controls. Any threshold cycle (Ct) value less than 35 was considered positive call. Relative miRNA expression levels were determined with the Ct method, normalized with the housekeeping genes Rnu6, rno-U87, which were geometrically averaged and reported as 2−Ct, where Ct = (target miRNA) − Ct (housekeeping). Expression was considered different, if fold change was >3. Different relative expression was found in 228 miRNA. Using the TargetScan database, we identified potential target genes of the miRNA influenced by quercetin. Thus we could link 35 miRNA to lipid metabolism, and 15 miRNA to carbohydrate metabolism. Two miRNA (rno-miR-19a, rno-miR-19b) have potential target genes involved in lipid as well as in carbohydrate metabolism. We conclude from the present study, that quercetin influences hepatic miRNA concentrations in a complex way and that parts of the multiple biological effects of quercetin might be mediated by this pathway. However, it is not possible at the moment to spec-