Fructus Ligustri Lucidi Extract Research Articles

Neuroprotective Effect of Fructus Ligustri Lucidi in Spinal Cord Neuron Injury during H2O2-Mediated Oxidative Stress

Oxidative stress-induced neuronal damage is a significant factor contributing to spinal cord injury. Although previous research has shown that Fructus Ligustri Lucidi (FLL) has neuroprotective benefits in SH-SY5Y, BV2, and PC12 cells, its impact on primary spinal cord neurons, which more accurately reflect the characteristics of central nervous system neurons, remains unexplored. This research investigated how FLL can protect rat primary spinal cord neurons from injury triggered by hydrogen peroxide (H2O2)-mediated oxidative stress. Cell viability, generation of reactive oxygen species (ROS), upregulation of inducible nitric oxide synthase (iNOS), activation of the Nrf2/HO-1 antioxidant pathway, and mitochondrial superoxide were assessed. Rat primary spinal cord neurons were not adversely affected by concentrations of FLL extract up to 100 μg/mL. Furthermore, FLL extract showed a significant protective effect against H2O2-induced neuronal toxicity at 10–100 μg/mL. Fluorescence-activated cell sorting analysis revealed that FLL extract inhibited H2O2-induced ROS generation in a dose-dependent manner. Immunocytochemistry and gene expression analysis confirmed that FLL extract reduced the overexpression of iNOS induced by H2O2 and enhanced the stimulation of the Nrf2/HO-1 pathway, crucial for antioxidant responses. In conclusion, FLL extract demonstrated neuroprotective effects on rat primary spinal cord neurons against the oxidative stress induced by H2O2. FLL extract effectively preserved cell viability, reduced ROS generation, suppressed iNOS overexpression, and activated the Nrf2/HO-1 antioxidant pathway. These results highlight the capacity of FLL extract as a neuroprotective agent against oxidative stress-related spinal neuron damage.

Optimization of Formulation Parameters in Preparation of Fructus ligustri lucidi Dropping Pills by Solid Dispersion Using 23 Full Experimental Design.

Oleanolic acid (OA) is an active ingredient of the traditional Chinese medicine (TCM) Fructus ligustri lucidi (FLL). Its clinical use is restricted because it is water-insoluble and has limited dosage forms of administration at present. Hence, the FFL dropping pills were prepared by the hot-melt method of solid dispersion technology. A 23 factorial design was used to examine the effects of the materials used to prepare the dropping pills (e.g., different ratios of PEG4000 and PEG6000, FLL extract loading, and percentage of Tween 80) on parameters such as dropping pill roundness, weight variation, and disintegration time. Moreover, 23 full factorial design was utilized to search for the optimal formulation for dissolution experiments. The results showed that the percentage of Tween 80 demonstrated significant effects on dropping pill roundness, weight variation, and disintegration time; FLL extract loading affected roundness and weight variation; and different ratios of PEG4000 and PEG6000 only affected disintegration time. The optimal formulation of the dropping pills released 70% of the drug after 30 min of dissolution release, which was faster than commercially available FLL Chinese medicines. Furthermore, the amount released was higher than that of commercially available formulations. In this study, a solid dispersion technique was used to successfully produce FLL dropping pills. In addition to improving the water insolubility of FLL and increasing the dissolution release percentage of the drug, we increased the application value of FLL and reduced the issues of traditional administration dosage forms.

Amelioration of Fructus Ligustri Lucidi and its phenol glycosides on hypercalciuria via stimulating PTH1R/PKA/TRPV5 signaling

BackgroundOur early studies performed on aged rats, ovariectomized (OVX) rats and diabetic mice, indicated the calciotropic role of Fructus Ligustri Lucidi (FLL), the fruit of Ligustrum lucidum Ait., in mediating calcium homeostasis which was partially attributed to its stimulation on renal calcium reabsorption. PurposeThis study aimed to explicate the underlying molecular mechanism and explore the potential bioactive ingredients in FLL. Study design and methodsThe OVX C57BL/6 J mice were orally administered with low (FL, 75 mg/kg), middle (FM, 225 mg/kg) or high (FH, 675 mg/kg) dose of extract of Fructus Ligustri Lucidi for 10 weeks. The biological properties of trabecular bone were measured by micro-CT and H&E staining. The molecular expression was assessed by immunoblotting and immunostaining. The potential active components were identified by cell membrane chromatography (CMC) and explored in renal tubular cells with Fluo-3/AM fluorescent staining to indicate intracellular calcium level. The male mice fed with high calcium diet (1.2% Ca) and orally treated with active components for 3 weeks. ResultsTreatment of OVX mice with FLL extract suppressed the elevation in urinary calcium level (FH, 0.081 ± 0.012, vs. OVX, 0.189 ± 0.038 mg/mg), and increased bone mineral density (FH, 62.41 ± 2.57, vs. OVX, 43.72 ± 8.43 mg/ccm) and percentage of trabecular bone area. It also decreased circulating PTH level (FH, 66.69 ± 10.94, vs. OVX, 303.50 ± 26.56 pg/ml) and up-regulated TRPV5 expression in renal cortex of OVX mice as well as enhanced the expression of PTH receptor (PTH1R) and the ratio of p-PKA/PKA. The PKA inhibitor H89 abolished the induction of serum, prepared from rats treated with FLL extract, on PKA/TRPV5 signaling in renal tubular cells. The CMC identified phenol glycosides, including salidroside and oleuropein, which increased intracellular calcium content, promoted expression of PTH1R and TRPV5 and ratio of p-PKA/PKA as well as decreased calcium excretion in urine of mice fed with high calcium diet. ConclusionSalidroside and oleuropein are major ingredients contributing to the anti-hypercalciuria effects of FLL via acting on PTH1R/PKA/TRPV5 signaling in kidney. Further translational research would be required.

Anti-diabetic activity of aqueous extract of Fructus Ligustri Lucidi in a rat model of type 2 diabetes

Purpose : To determine the anti-diabetic activity of an aqueous extract of Fructus Ligustri Lucidi (FLL) in a rat model of type 2 diabetes (T2D). Method : Body weight (BW), food intake (FI), fasting blood glucose (FBG), glucose tolerance testing and insulin tolerance testing were used to determine the anti-diabetic activity of an aqueous extract of Fructus Ligustri Lucidi (FLL) in a rat model of streptozotocin-induced T2D. Anti-oxidant activity and oxidative stress were assessed by superoxide dismutase (SOD) and malondialdehyde (MDA) analyses, respectively. Results : Following FLL extract treatment, diabetic rat BW increased, while FI and FBG levels decreased. FLL extract increased glucose tolerance and decreased insulin tolerance. Following treatment with 300 and 600 mg/kg FLL extract, MDA levels reached 13.5 ± 0.9 nmol/ml and 13.8 ± 1.1 nmol/ml, respectively. Compared to MDA levels of 19.5 ± 1.1 nmol/ml in diabetic control group, MDA levels was decreased by 30.8 % and 29.0 % after the treatment with 300 and 600 mg/kg FLL extract, respectively, indicating alleviated oxidative stress. Conclusion : The results show that aqueous FLL extract has the potential to alleviate T2D, resulting in reduced FI and FBG, and increased BW. The anti-diabetic activity of FLL extract on T2D may be relevant to reduced oxidative stress burden. Keywords : Fructus Ligustri Lucidi, Type 2 diabetes, Anti-oxidant activity, Glucose tolerance, Insulin tolerance, Anxiety-like behaviour

Antioxidant Effect of Fructus Ligustri Lucidi Aqueous Extract in Ovariectomized Rats Is Mediated through Nox4-ROS-NF-κB Pathway.

Purpose: This study is designed to explore whether Fructus ligustri lucidi (FLL) exhibits antioxidant effect in ovariectomized (OVX) rats, and to identify the signaling pathway involved in this process.Methods: OVX rats were treated with FLL aqueous extract (3.5 g/kg) for 12 weeks. Serum, uteri, and tibias were harvested from the rats and the levels of total antioxidant capacity (TAC), nitric oxide (NO), malondialdehyde (MDA), 8-hydroxy-desoxyguanosine (8-OHdG), and superoxide dismutase (SOD) were determined. Changes in the levels of NF-κB-p65, phosphorylation of NF-κB-p65 (NF-κB-pp65), NF-κB inhibitor alpha (IκBα), phosphorylation of IκBα (p-IκBα), and NADPH oxidase 4 (Nox4) in uteri and tibias were determined by western blot, immunofluorescent and immunohistochemical analysis, respectively. In addition, the expression of cytochrome C (Cyto-C) and B-cell lymphoma-2 (Bcl-2) were determined in the tibias of rats. Histopathological changes in the bones were evaluated by hematoxylin-eosin staining. Bone mineral density (BMD) was determined in rat femurs by dual X-ray absorptiometry.Results: Treatment of OVX rats with FLL aqueous extract improved redox homeostasis by increasing the levels of TAC and NO as well as decreasing the levels of MDA and 8-OHdG in serum, tibias, and uteri. Further, FLL extract also downregulated the expression of Nox4, NF-κB-p65, NF-κB-pp65, and p-IκBα in the uteri and tibias. Furthermore, administration of FLL–OVX rats increased Bcl-2 expression and prevented cytoplasmic release of mitochondrial Cyto-C in the tibias. In addition, FLL treatment also improved bone microstructure and increased cortical bone thickness as well as increased BMD values in the femurs of OVX rats.Conclusions: FLL treatment may suppress oxidative stress response in OVX rats via regulating the Nox4/ROS/NF-κB signaling pathway. These results suggest the potential of using FLL as a natural antioxidant agent in preventing the development of osteoporosis.

Effect of the water fraction isolated from Fructus Ligustri Lucidi extract on bone metabolism via antagonizing a calcium-sensing receptor in experimental type 1 diabetic rats.

Our previous studies have demonstrated that the extract of Fructus Ligustri Lucidi (FLL) can maintain in vivo calcium homeostasis in aged and ovariectomized rats. This study was designed to elucidate the action of water fraction isolated from the FLL extract on bone metabolism and a calcium-sensing receptor (CaSR) in parathyroid glands and kidneys of diabetic rats. The streptozotocin-induced diabetic rats were treated with vehicle, FLL extract, and the water fraction (WF) isolated from the FLL extract for 4 weeks. Treatment with WF dramatically increased the serum levels of both calcium and parathyroid hormone and reduced urinary calcium excretion in diabetic rats as well as improved the pathological changes of trabecular bone as shown by the increased BA/TA, BMD/BV, and BV/TV. The mRNA expression of the calcium-binding protein 9k and protein expression of a vitamin D receptor (VDR) and plasma membrane Ca-ATPase in duodenum were significantly increased in diabetic rats after treatment with WF, which reduced the expression of CaSR in parathyroid gland and kidney as well as inhibited the up-regulation of VDR and 25-hydroxyvitamin D-24 hydroxylase expressions in the kidney of diabetic rats. This study reveals that the water fraction may be an active component of the FLL extract that exerts beneficial effects on improving bone metabolism via regulating vitamin D metabolism in kidney and vitamin D-dependent calcium transporters in duodenum as well as modulating the expression of CaSR in the parathyroid gland and kidneys.

Fructus Ligustri Lucidi ethanol extract inhibits osteoclastogenesis in RAW264.7 cells via the RANKL signaling pathway.

Fructus ligustri Lucidi (FLL) is the fruit of Ligustrumlucidum Ait and a traditional Chinese medicine, primarily known for its role in osteoporosis prevention and treatment. The present study aimed to elucidate the effect and underlying mechanism of action of ethanol extract of FLL on osteoclast differentiation and bone resorption, and to identify the active compounds within it. RAW264.7 murine monocyte/macrophage cells were stimulated with the receptor activator of nuclear factor κB ligand (RANKL) to induce osteoclast differentiation invitro. The present study demosntrated that FLL extract and its two primary components, oleanolic acid (OA) and ursolic acid (UA), significantly suppressed RANKL‑induced tartrate resistant acid phosphatase (TRAP) activity and multinucleate osteoclast formation without inducing cytotoxicity; however, no effect was observed on the apoptosis of mature osteoclasts. Additionally, RANKL‑induced mRNA expression levels of the key transcription factors, tumor necrosis factor receptor associated factor‑6, nuclear factor of activated Tcell‑c1 and c‑Fos, and the osteoclast markers, TRAP, cathepsin K and matrix metalloproteinase‑9 were suppressed by FLL, OA and UA. However, no effect was observed on RANKL‑induced mRNA expression levels of Src. These results demonstrated that FLL may inhibit osteoclastogenesis in RAW264.7 cells via RANKL signaling pathways. OA and UA are active compounds in inducing this effect; however, their specific roles remain to be elucidated.

Effect of the Fructus Ligustri Lucidi extract and its monomers quercetin and oleanolic acid on the adhesion and migration of melanocytes and intracellular actin.

The present study aimed to investigate the effects of the Fructus Ligustri Lucidi (FLL) extract and its monomers quercetin and oleanolic acid on the adhesion and migration of human epidermal melanocytes (MCs) and intracellular actin. The human epidermal MCs were cultured and identified. The cells were treated with different concentrations of FLL extract, quercetin and oleanolic acid. The adhesion and migration abilities of the cells were determined by the fibronectin-coated culture experiment and Transwell assay, respectively. The structure and distribution of intracellular actin were observed by confocal laser microscopy, with semi-quantitative analysis. Results showed that compared with the control group, 0.0375-0.3 mg/ml of the FLL extract and 40 µM quercetin significantly improved the adhesion rate of MCs (P<0.05). The numbers of MCs permeating the microporous membrane in the 0.15 mg/ml FLL extract and 12 µM oleanolic acid groups were 43.7 and 30.3, respectively, significantly higher compared to the control group (P<0.01). In the control group, the intracellular actin was less, and the stress fiber structure was not clear. In the 0.15 mg/ml FLL extract, 12 µM oleanolic acid and 40 µM quercetin groups, there were numerous bunched stress fibers, indicating the aggregation of filamentous fibrous actin. The mean optical densities of actin expression in the 0.15 mg/ml FLL extract, 12 µM oleanolic acid and 40 µM quercetin groups were significantly higher compared to the control group (P<0.05). The FLL extract has a significant stimulatory effect on the adhesion and migration of human epidermal MCs. The mechanism may be associated with the promotion of intracellular actin cytoskeleton aggregation.

Combination therapy of Chinese herbal medicine Fructus Ligustri Lucidi with high calcium diet on calcium imbalance induced by ovariectomy in mice

Context: Our previous biological study demonstrated that Fructus Ligustri Lucidi (FLL), the fruit of Ligustrum lucidum Ait. (Oleaceae), could be used to maintain calcium balance and prevent age-related osteoporosis since it effectively decreased calcium loss and increased calcium retention in rats.Objective: This study investigates the combination effect of the Chinese herbal medicine FLL and a high calcium diet on calcium imbalance induced by ovariectomy in mice.Materials and methods: The ovariectomized (OVX) mice were orally treated with vehicle, FLL extract (700 mg/kg), milk powder (5 g/mice) fortified with calcium (1.0% Ca) and the combination of FLL with milk powder. After 6 weeks of treatment, urine, serum, and tibia were preserved for biochemical analysis and kidneys were taken for gene expression analysis.Results: The combination treatment of FLL and a high calcium diet significantly increased bone calcium content (6.80 ± 0.34 mg) by 22% (p < 0.05) and decreased urine calcium excretion (0.099 ± 0.009 mg/mg) by 62% (p < 0.01) as compared with those of the OVX group (bone Ca, 5.57 ± 0.31 mg; urine Ca/Cr, 0.261 ± 0.017 mg/mg). The mRNA expression of renal calcium-binding protein-9k (CaBP-9k) and calcium-sensing receptor (CaSR) in combination treatment group was significantly up-regulated and down-regulated, respectively, as compared with those of the OVX group.Conclusion: The beneficial effects of this combination therapy on calcium balance of OVX mice were, at least partially, attributed to its regulation on mRNA expression of CaBP-9k and CaSR in kidney.

Protective effects of water fraction of Fructus Ligustri Lucidi extract against hypercalciuria and trabecular bone deterioration in experimentally type 1 diabetic mice

Ethnopharmacological relevanceFructus Ligustri Lucidi (FLL), the fruit of Ligustrum lucidum Ait, is a commonly prescribed herb to nourish the endocrine and renal systems and to strengthen the bones in Traditional Chinese Medicine. This study was aimed to determine the effects of water fraction of FLL ethanol extract (WF-EE) on urinary calcium excretion and trabecular bone properties in type 1 diabetic mice. Materials and methodsThe DBA/2J mice with type 1 diabetes induced by streptozotocin injection were orally administered with WF-EE. After 6 weeks of treatment, the level of biomarkers, including serum calcium, parathyroid hormone (PTH), and fibroblast growth factor-23 (FGF-23) and urine calcium, was measured. Micro-CT was performed to detect trabecular bone properties of the proximal tibial metaphysis. The expression of active calcium transporting proteins in kidney and duodenum was determined by RT-PCR, immunoblotting and immunostaining. ResultsType 1 diabetes induced hypercalciuria and trabecular bone deterioration. The WF-EE could significantly inhibit hypercalciuria and ameliorate the micro-structure of trabecular bone as well as increase serum PTH and FGF-23 levels in type 1 diabetic mice. The gene expressions of active calcium transporting proteins in duodenum were up-regulated, and the gene and protein expressions of calcium-sensing receptor (CaSR) in kidney were dramatically down-regulated in diabetic mice in response to the treatment with WF-EE. ConclusionsThe present study demonstrated the protective effects of the water fraction of Fructus Ligustri Lucidi ethanol extract against hypercalciuria and trabecular bone deterioration in experimentally type 1 diabetic mice, and the underlying mechanism may be attributed to its regulations on duodenal calcium transporting proteins and renal CaSR.

Fructus Ligustri Lucidi (FLL) ethanol extract increases bone mineral density and improves bone properties in growing female rats

Osteoporosis is a chronic disease affecting millions of people worldwide. It is generally accepted that acquisition of a high peak bone mass (PBM) early in life can reduce the risk of osteoporosis later in life. The aims of this study were to investigate the effects of Fructus Ligustri Lucidi (FLL) ethanol extract on bone mineral density and its mechanical properties in growing female rats and to explore the underlying mechanisms. The rats were given different doses of FLL extract mixed with AIN-93G formula (0.40, 0.65 and 0.90 %), and a group given AIN-93G diet treatment only was used as control. The intervention lasted for 16 weeks until the animals were about 5 months old, the time when the animals almost reach their PBM. Our results showed that FLL treatment increased bone mineral density and improved bone mechanical properties in the growing female rats in a dose-dependent manner. In addition, FLL treatment significantly decreased the serum bone-resorbing marker, CTX-I, while significantly increasing serum 25(OH)D3 and thereby increasing Ca absorption and Ca retention. Intriguingly, both in vivo and in vitro results demonstrated that FLL treatment could reduce the RANKL/OPG ratio. In conclusion, FLL ethanol extract exerted beneficial effects on peak bone mass acquisition and the improvement of bone mechanical properties by favoring Ca metabolism and decreasing the RANKL/OPG ratio.

Therapeutic effect of aqueous extract from Ecliptae herba on bone metabolism of ovariectomized rats

Ecliptae herba (EH) has long been used in China to strengthen bones. Accumulating evidence indicates that EH may have antiosteoporotic effects. The aim of this study was to evaluate the effects of aqueous EH extract (EHE) on rats that had osteoporosis-like features induced by ovariectomy, using aqueous Fructus Ligustri Lucidi extract as positive control agent. Three-month-old female rats that underwent ovariectomy were treated with EHE (1.4 g/kg per day). After 12 weeks, bone mineral density and bone histomorphometric indices of tibiae were measured. Protein and messenger RNA expressions of osteoprotegerin and receptor activator of nuclear factor κ-B ligand (RANKL) in tibiae were evaluated by immunohistochemistry and in situ hybridization. In addition, serum concentrations of osteocalcin, interleukin-1β, interleukin-6 (IL-6), calcitonin (CT), and parathyroid hormone were determined by enzyme-linked immunosorbent assay. EHE treatment prevented body weight gain and loss of uterine wet weight in ovariectomized rats. It remarkably increased bone mass in ovariectomized rats compared with ovariectomized controls. EHE treatment significantly down-regulated RANKL expression in tibiae from ovariectomized rats compared with controls; however, it had no significant effect on osteoprotegerin expression. In addition, EHE treatment significantly reduced serum IL-6 levels and remarkably increased CT levels but had no effect on parathyroid hormone. EHE increases bone mass in ovariectomized rats by inhibiting bone loss: down-regulated RANKL expression in tibiae and IL-6 level in serum, and up-regulated CT level in serum. This suggests that EHE may be developed as an alternative therapeutic agent for osteoporosis induced by postmenopause.

Effect of Supplemental Fructus Ligustri Lucidi Extract on Methane Production and Rumen Fermentation in Sheep

The effect of addition of Fructus Ligustri Lucidi Extract (FLLE) on methane production and rumen fermentation in sheep was investigated in this study. FLLE addition levels were 0, 1, 2, 3 and 4 g/L of ruminal fluid. Ruminal fluid was removed from four sheep, mixed with phosphate buffer (1:2) and incubated (30 ml) anaerobically at 38°C for 24-h with or without FLLE. Results indicated that addition of FLLE linearly decreased methane production (p&lt;0.05), linearly decreased ammonia concentration (p&lt;0.05), and acetate to propionate ratio (p&lt;0.05). Data of numbers of methanogens, fungi, ciliate and cellulolytic indicated that they were linearly decreased by addition of FLLE (p&lt;0.05). Addition of FLLE at all dosages have no effect on rumen pH value, acetate molar concentration and butyrate molar concentration (p&gt;0.05). Addition of FLLE linearly decreased count of F. succinogenes (p&lt;0.05). In conclusion, these results of 24-h in vitro batch culture indicated that FLLE has the ability to inhibit methane production in the rumen, stimulate rumen fermentation and suppress fiber degrading bacteria, F. succinogenes. Abbreviations: FLLE=Fructus Ligustri Lucidi extract; IVDDM=in vitro digestibility of dry matter.

Effect of several supplemental Chinese herbs additives on rumen fermentation, antioxidant function and nutrient digestibility in sheep

Two experiments were carried out in this study. Experiment 1 was conducted to examine the effects of several supplemental Chinese herbs on antioxidant function and slaughtered body weight in sheep. Results indicated that Fructus Ligustri Lucidi supplementation improved the blood antioxidant function [higher concentration of glutathione reductase (GR), superoxide dismutase and lower concentration of malondialdehyde] and slaughtered body weight in sheep (p < 0.05). Experiment 2 was conducted to investigate the effect of Fructus Ligustri Lucidi extract (FLLE) on rumen fermentation and nutrient digestibility in sheep. Four levels of FLLE treatments, i.e. 0, 100, 300 and 500 mg/kg dry matter (DM), were used in this part. Addition of FLLE at 300 or 500 mg/kg DM increased total volatile fatty acid (VFA) concentration and propionate proportion, decreased ammonia-N concentration in the ruminal fluid, reduced blood urea nitrogen concentration at 2, 4, 6 and 8 h after morning feeding (p < 0.05). Addition of FLLE at all dosages had no effect on ruminal pH value and acetate concentration at all sampling time points in sheep (p > 0.05). Dynamic degradation coefficient c of maize DM was significantly increased by supplementing FLLE at 300 or 500 mg/kg DM (p < 0.05). Fructus Ligustri Lucidi extract addition had no effect on degradation coefficients a, b, c of DM and nitrogen of soybean meal; a, b of maize DM; a, b, c of maize nitrogen; and a, b, c of neutral detergent fibre (NDF) and acid detergent fibre (ADF) of Chinese wildrye (p > 0.05). Addition of FLLE at 300 or 500 mg/kg DM increased DM and organic matter digestibility of diet (p < 0.05). Fructus Ligustri Lucidi extract addition had no effect on digestibility of diet's NDF, ADF and crude protein (p > 0.05). From the aforementioned results, it is indicated that FLLE improved antioxidant status and slaughtered body weight. Fructus Ligustri Lucidi extract addition has capability to modulate rumen fermentation, increase the maize degradation rate, total volatile fatty acid concentration and propionate proportion in sheep.

Improvement of bone properties and enhancement of mineralization by ethanol extract of Fructus Ligustri Lucidi

Fructus Ligustri Lucidi (FLL), a kidney-tonifying Chinese herb, was shown to regulate Ca balance in ovariectomized (OVX) rats in our previous study. This study investigated whether it could improve bone properties in aged normal and OVX rats and increase osteoblastic differentiation in rat osteoblast-like UMR-106 cells. Ten-month-old aged rats underwent sham-operation or ovariectomy, were orally administered with FLL extracts or its vehicle and fed with diets containing different levels of Ca (LCD, 0.1 % Ca; MCD, 0.6 % Ca; HCD, 1.2 % Ca) for 12 weeks. Ovariectomy induced bone loss at multiple-sites of both tibia and femur in all rats being studied. FLL extract increased bone mineral density and bone mineral content at both tibial and femoral diaphysis as well as the lumbar vertebra (LV-2) in rats fed either LCD or MCD. In addition, FLL increased biomechanical strength of the tibial diaphysis in these rats. Combination of FLL and high-Ca diet significantly improved bone mass of cortical and trabecular bone at appendicular bones and LV-2 and decreased bone loss associated with ovarietomy and low-Ca feeding. Treatment of UMR-106 cells with FLL extracts accelerated the formation of calcified matrix and increased extracellular Ca and P depositions in time- and dose-dependent manner. The level of mineralization reached a maximum by 6 d incubation at the dosage of 10 mug FLL extract/ml. Our study indicated that FLL extract could improve bone properties in aged rats possibly via its direct action on osteoblastic cells by enhancement of the mineralization process.

Improvement of Ca balance by Fructus Ligustri Lucidi extract in aged female rats

This study aimed to demonstrate and delineate the mechanism of action of Fructus Ligustri Lucidi (FLL) involved in improving Ca balance in aged female rats. FLL could enhance the apparent Ca absorption rate and reduce Ca excretion, via its actions on modulating the levels of calciotropic hormones and CaBPs expression. Fructus Ligustri Lucidi (FLL) is a herb classically used for the treatment of age-related symptoms in China. The present study aimed to determine if FLL could improve calcium balance in aged female rats and delineate its mechanism of action in vivo. Aged Sprague-Dawley rats (11 months of age) were orally administered with ethanol extract of FLL or its vehicle and fed with diets containing different levels of Ca (low Ca diet, LCD, 0.1% Ca; medium Ca diet, MCD, 0.6% Ca; high Ca diet, HCD, 1.2% Ca) for 12 weeks. Serum, urine and feces were collected for biochemical markers and Ca balance determination. mRNA expressions of calcium binding proteins (CaBPs) were determined by real time RT-PCR. The effects of diets and herb were analyzed by both one-way and two-way ANOVA. FLL significantly reduced fecal Ca excretion and induced apparent Ca absorption rate in rats fed with MCD and HCD. FLL increased serum 1,25(OH)(2)D(3) level and duodenal CaBP-9k mRNA expressions in all rats. Renal CaBP-28k mRNA expressions were induced in rats fed with MCD and HCD upon FLL treatment. Our study demonstrated that FLL improved Ca balance in aged female rats by increasing serum 1,25 (OH)(2)D(3) level and vitamin D-dependent CaBPs expression.

Effects of Fructus Ligustri Lucidi Extract on Bone Turnover and Calcium Balance in Ovariectomized Rats

The aim of this study was to evaluate the effect of Fructus Ligustri Lucidi (FLL), a kidney-tonifying Chinese herbal medicine, on the biochemical markers of bone turnover, calcium metabolism and balance in osteoporotic rat model developed by ovariectomy. Four weeks after surgical operation, animals were randomly assigned to one of the four treatments for 14 weeks: sham-operated control treated with vehicle (sham, n=8), ovariectomized group treated with vehicle (OVX, n=8), OVX group treated with 17beta-estradiol (E(2), n=10, 2 microg/kg/d) and OVX group treated with FLL extracts (FLL, n=10, 550 mg/kg/d). Serum osteocalcin and urinary deoxypyridinoline levels were upregulated in rats in response to OVX, suggesting that the bone turnover rate was accelerated in these animals. Treatment of OVX rats with FLL extract could prevent OVX-induced increase in bone turnover by suppression of both serum osteocalcin (p<0.05, vs. OVX) and urinary deoxypyridinoline (p<0.05, vs. OVX) levels. In addition, FLL extract could prevent OVX-induced loss of calcium in rats by increasing the intestinal calcium absorption rate (p<0.01, vs. OVX), suppressing urinary Ca excretion (p<0.05, vs. OVX) as well as increasing bone calcium content (p<0.05, vs. OVX). Our study is the first to report that FLL can modulate bone turnover and calcium balance in OVX rats and it might be a potential candidate for prevention and treatment of postmenopausal osteoporosis.