A traditional Chinese herbal preparation, Er-Zhi-Wan, prevent ovariectomy-induced osteoporosis in rats

Ethnopharmacological relevance

Er-Zhi-Wan (EZW), a classic Chinese formulation, which contains Fructus Ligustri Lucidi (FLL) and Herba Ecliptae (HE). EZW is widely used to prevent and treat various kidney diseases for its actions of nourishing the kidney and strengthening tendon and bone. Although recent reports indicate that EZW restrains osteoclastic bone resorption, its effects on the protection against define OVX-induced bone loss in mature rats have not been systematically investigated.

Materials and methods

Sixty 3-month-old female Sprague-Dawley rats were randomly assigned into sham-operated group (Sham) and five OVX subgroups, OVX with vehicle (OVX); OVX with Estradiol Valerate (EV, 0.4 mg/kg body weight/day); OVX with EZW of graded doses (9.0, 4.5, or 2.25 g/kg/day). Daily oral administration of EV and EZW on 5th week for 26 weeks. Bone turnover markers (Serum alkaline phosphatase (ALP), bone-specific alkaline phosphatase (BALP), osteocalcin (OCN), deoxypyridinoline (DPD)), other parameters, including serum calcium (S-Ca), serum phosphorus (S-P), urine calcium (U-Ca), phosphorus (U-P), and bone mineral density (BMD) of the femur, 4th lumbar vertebra and tibia, bone biomechanical properties and trabecular microarchitecture parameters were measured.

Results

Administration of EZW could significantly prevent ovariectomy-induced bone loss, biomechanical reduction, deterioration of trabecular microarchitecture and the body weight gain without affecting the weight of the uterus, and increased S-Ca, S-P levels, decreased level of bone turnover markers and U-Ca, U-P levels in ovariectomized rats.

Conclusion

The present study indicated that EZW had a definite antiosteoporotic effect without hyperplastic effect on uterus, and it might be a potential alternative medicine for treatment of postmenopausal osteoporosis.     

Comparison of chemical compositions and osteoprotective effects of different sections of velvet antler

Ethnopharmacological relevance

Velvet antlers (VA) have been claimed for centuries to have numerous medical benefits including strengthen bones. To investigate and compare the anti-osteoporotic activities from different sections of VA.

Materials and methods

Fresh VA prepared from farmed sika deers (Cervus nippon) was divided into upper (VAU), middle (VAM), and basal (VAB) sections. The chemical constituents and anti-osteoporotic effect of different sections from VA were evaluated using ovariectomized rats.

Results

Levels of water-soluble extracts, diluted alcoholic extract, amino acids, testosterone, insulin-like growth factor (IGF)-1and testosterone plus estradiol significantly differed among the different sections. Levels of these constituents were significantly higher in the upper section than in the basal section. Moreover, levels of testosterone and IGF-1 of the VAM were also significantly higher than those of the VAB. Calcium level increased downward from the tip with statistical significance. The strength of vertebrae increased in all VA-treated groups compared to the control, but only treatment with VAU and VAM increased the strength of the femur and the microarchitecure of the trabecular bone. Alkaline phosphatase levels of VAU- and VAM-treated groups significantly decreased, but osteocalcin did not significantly change. Moreover, VAU and VAM dose-dependently increased proliferation and mineralization of MC3T3-E1 cells.

Conclusion

Our study provides strong evidence for the regional differences in the effectiveness of velvet antler in treating osteoporosis. However, further studies are needed to elucidate the bioactive chemical constituents associated with the anti-osteoporotic effects of velvet antler.     

Osteoprotective effect of extract from Achyranthes bidentata in ovariectomized rats

Aim of the study

A Chinese herbal drug, root of Achyranthes bidentata showed a potent inhibitory activity on bone resorption induced by parathyroid hormone (PTH) in a bone organ culture using neonatal mouse parietal bones. The present study is to clarify the fractions responsible for the activity and further explore the osteoprotective effect of the fraction in vivo.

Materials and methods

The hexane, ethyl acetate (EtOAc), n-butanol (n-BuOH) and water soluble fractions of methanol extract of the root of Achyranthes bidentata were prepared and screened for their anti-bone resorption activity using the bone organ culture system. The n-BuOH soluble fraction was further administered orally at doses of 25, 50 and 100 mg/(kg day) to ovariectomized (OVX) rats. The analyses of the rat body weight, serum estradiol (E2), total cholesterol and triglyceride levels, uteri weight and measurement of bone mineral density (BMD) were conducted.

Results

The EtOAc and n-BuOH fractions showed the most potent inhibitory activity on PTH-induced bone resorption. Further research using OVX rat model revealed that the n-BuOH fraction significantly prevented BMD loss due to OVX operation. While, the uteri weight and serum estradiol (E2), total cholesterol and triglyceride levels displayed no differences compared with those of control group (OVX rats), suggesting the n-BuOH fraction should have no estrogen-like side effects.

Conclusions

The results reveal that the n-BuOH soluble fraction of the root of Achyranthes bidentata is effective at preventing bone loss in OVX rats and has a great potential as an alternative tool for the treatment of osteoporosis.     

Hachimijiogan (Ba-Wei-Di-Huang-Wan), a herbal medicine,improves unbalance of calcium metabolism in aged rats

Aim of the study

Aged animals as well as elderly humans commonly exhibit calcium (Ca) shortage because of increased Ca excretion into urine and decreased intestinal Ca absorption, which induce elevation of serum PTH levels to maintain serum Ca levels between a normal physiological range. The most important organ that regulates this Ca homeostasis is the kidney. Hachimijiogan (HJG), a traditional herbal medicine in Japan and China, has been used for treating clinical diseases associated with kidney dysfunctions in elderly humans. However, the mechanisms of its pharmacological actions remain to be understood poorly. The present study was designed to examine whether HJG improves age-related unbalance of Ca metabolism at the systemic level using aged rats.

Materials and methods

HJG was administered to 21-month-old aged rats for 3 months, and several parameters associated with Ca metabolism in serum and urine were measured.

Results

Although HJG as well as aging itself did not affect serum Ca levels compared to young (11-week-old) rats, HJG improved increase in urinary Ca excretion and elevation of serum parathyroid hormone (PTH) levels in aged rats. However, HJG did not improve marked reduction of intestinal Ca absorption in aged rats.

Conclusion

HJG showed regulating action for age-related unbalance of Ca metabolism at the systemic level. This finding would provide useful information for treating age-related several disorders associated with Ca unbalance.     

Effects of total lignans from Eucommia ulmoides barks prevent bone loss in vivo and in vitro

Ethnopharmacological relevance

The present study systematically investigate the in vivo and in vitro effect of total lignans (TL) extracted from Eucommia ulmoides Oliv. barks on bone formation using ovariectomy rat model and primary cultures of rat osteoblasts.

Materials and methods

Eighty 3-month-old female Sprague-Dawley rats were used and randomly assigned into sham-operated group (SHAM) and five ovariectomy (OVX) subgroups, i.e. OVX with vehicle (OVX); OVX with 17α-ethinylestradiol (E₂, 25 μg/kg/day); OVX with TL of graded doses (20, 40, or 80 mg/kg/day). The treatment began 4 weeks after the surgery and lasted for 16 weeks. in vitro experiments were performed to determine the potential mechanisms of the anti-osteoporotic effect of TL.

Results

Treatment with TL significantly prevent OVX-induced decrease in biomechanical quality of femur such as maximum stress and Young?s modulus. The mechanical changes were associated with the prevention of a further BMD decrease or even with some improvements in microarchitecture. TL inhibited BMD decrease in the femur caused by OVX, which was accompanied by a significant decrease in skeletal remodeling, as was evidenced by the decreased levels of the bone turnover markers. μCT analysis of the femoral metaphysis showed how to prevent the deterioration of trabecular microarchitecture. TL induced primary osteoblastic cell proliferation and differentiation, inhibition of osteoclastogenesis through an increase in osteoprotegrin (OPG) and a decrease in NF-κB ligand (RANKL) expression in vitro.

Conclusions

We concluded that TL treatment can effectively suppress the loss of bone mass induced by OVX and in vitro evidence suggests this could be through actions on both osteoblasts and osteoclasts.     

The osteoprotective effect of Radix Dipsaci extract in ovariectomized rats

Aim

The objective of the present study was to systematically evaluate the effects of Radix Dipsaci extract (RDE) on postmenopausal osteoporosis.

Materials and methods

OVX or sham operations were performed on sixty 3-month-old virgin Sprague–Dawley rats that were divided into six groups: sham control group (sham, n = 10); OVX control group (OVX, n = 10); 17β-estradiol treatment group (E2, n = 10); three Radix Dipsaci extract treatment groups RDE100 (n = 10), RDE300 (n = 10) and RDE500 (n = 10). The treatment began 4 weeks after the surgery and lasted for 16 weeks. Bone mass, bone turnover and strength were analyzed by DEXA, biochemical markers and three-point bending test. The trabecular bone microarchitecture was evaluated by MicroCT.

Results

16 weeks treatment of RDE slowed down the body weight gain and prevented the loss of bone mass induced by the OVX. The prevention effect on bone loss was due to altering the rate of bone remodeling, which could be inferred from the decreased level of bone turnover markers, such as serum ALP, OC and urinary DPD. The changes of urinary calcium and phosphorus excretion provided the same evidence. The treatment could also enhance the bone strength and prevent the deterioration of trabecular microarchitecture.

Conclusions

Our study provides evidence that Radix Dipsaci extract will have potential to be used for treatment of postmenopausal osteoporosis.     

Supplementing with Opuntia ficus-indica Mill and Dioscorea nipponica Makino extracts synergistically attenuates menopausal symptoms in estrogen-deficient rats

Ethnopharmacological relevance

Prickly pear cactus grown in Korea (Opuntia ficus-indica Mill, KC) and Buchema (Dioscorea nipponica Makino, B) have been traditionally used in East Asia and South America to treat various metabolic diseases. The aim of the present study was to determine whether the extracts of KC, B, and KC+B can prevent the impairments of energy, glucose, lipid and bone homeostasis in estrogen-deficient ovariectomized (OVX) rats and to explore their mechanisms.

Materials and methods

OVX rats were divided into 4 groups and fed high fat diets supplemented with either 3% dextrin (control), 3% KC, 3% B or 1.5% KC+1.5% B. Sham rats were fed 3% dextrin. After 12 weeks of diet consumption, energy, lipid, glucose and bone metabolisms were analyzed and Wnt signaling in the femur and hepatic signaling were determined.

Results

OVX impaired energy, glucose and lipid metabolism and decreased uterine and bone masses. B and KC+B prevented the decrease in energy expenditure, especially from fat oxidation, in OVX rats, but did not affect food intake. KC+B and B reduced body weight and visceral fat levels, as compared to the OVX-control, by decreasing fat synthesis and inhibiting FAS and SREBP-1c expression. KC+B and B prevented the increases in serum lipid levels and insulin resistance by improving hepatic insulin signaling (pIRS→pAkt→pGSK-3β). KC and KC+B also prevented decreases in bone mineral density (BMD) in the femur and lumbar spine in OVX rats. This was related to decreased expressions of bone turnover markers such as serum osteocalcin, alkaline phosphatase (ALP) and bone-specific ALP levels, and increased serum P levels. KC and KC+B upregulated low-density lipoprotein receptor-related protein 5 and β-catenin in OVX rats, but suppressed the expression of dickkopf-related protein 1. B alone improved energy, lipid and glucose homeostasis, but not bone loss, whereas KC alone enhanced BMD, but not energy, lipid or glucose homeostasis.

Conclusion

KC+B synergistically attenuated impairments of bone, energy, lipid and glucose metabolism by OVX, suggesting potential efficacy of the combination for alleviating menopausal symptoms.     

The anti-osteoporotic effect of velvet antler polypeptides from Cervus elaphus Linnaeus in ovariectomized rats

Ethnopharmacological relevance

The deer velvet antler is well known for its traditional medicinal value, and is widely used in the clinic. It is recorded in the Compendium of Materia Medica that the deer velvet antler replenishes vital essence and strengthens the bone.

Aim of the study

The goal of this study was to investigate the anti-osteoporotic effect of total velvet antler polypeptides from Cervus elaphus Linnaeus (TVAPL) on ovariectomized rats (OVX), and their possible mechanism of the action.

Materials and methods

Wistar rats were divided into five groups: sham-operated group, OVX group, and OVX rats treated with 20, 40, or 60 mk/kg TVAPL for 12 weeks. Calcium and phosphorus levels, bone weight coefficient (BWC), bone mineral density (BMD), and bone mineral content (BMC) were evaluated. The MTT assay was used to measure the activities of interleukin-1 (IL-1) and interleukin-6 (IL-6). In addition, cartilage cells and osteoblast-like cells were exposed to TVAPL, natural velvet antler polypeptides (nVAP), and synthetic velvet antler polypeptides (sVAP), to determine their effects on cell proliferation using the tritiated thymidine incorporation assay. Finally, the enzyme-linked immunosorbent assay was used to determine the effects of nVAP and sVAP on cytokines related to bone metabolism.

Results

The administration of TVAPL for 12 weeks significantly reversed osteoporosis in OVX rats, thereby improving the BWC, BMD, BMC, and bone microarchitecture. IL-1 and IL-6 were significantly activated in the OVX group, and their activation was inhibited by TVAPL. In addition, nVAP and sVAP promoted the proliferation of cartilage and osteoblast-like cells (p<0.01 or p<0.001), and inhibited the secretion of IL-1α from THP-1 monocytic cells in vitro.

Conclusion

These results suggest that TVAPL are effective in preventing bone loss in OVX rats. The effect of TVAPL on osteoporosis is due to inhibition of IL-1 and IL-6 by nVAP, and promotion of mitosis. sVAP has similar bioactivity as nVAP. Thus, both TVAPL and sVAP may be potential therapeutic agents for the treatment of postmenopausal osteoporosis.     

Effect of the combinatory mixture of Rubus coreanus Miquel and Astragalus membranaceus Bunge extracts on ovariectomy-induced osteoporosis in mice and anti-RANK signaling effect

Ethnopharmacological relevance

Postmenopausal osteoporosis is one of the most common disorders in women after menopause, which is linked to an estrogen deficiency and characterized by an excessive loss of trabecular bone. Rubus coreanus and Astragalus membranaceus have been used for their various pharmacological properties in Asia as a traditional medicine. The present study evaluated the anti-osteoporotic effects of the optimal combination of Rubus coreanus and Astragalus membranaceus in 7:3 mixture (RAM) in ovariectomized (OVX) mice by investigating bone biomechanical properties and the serum levels of TNF-α, osteocalcin, RANKL, OPG, and RANK–RANKL signal-related osteoclast differentiation markers.

Materials and methods

A total of 36 mature female outbred ICR (Institute of cancer research) strain mice (7 weeks) were divided into 6 groups with 7 mice in each group as follows: (1) Sham-operated control mice (Sham) received daily oral phosphate-buffered-saline (PBS) of equal volumes through gavage. (2) OVX mice received a daily oral gavage of PBS (OVX). (3) OVX mice were treated daily with 50 mg/kg b.w./day of RAM (4) with 100 mg/kg b.w./day of RAM or (5) with 200 mg/kg b.w./day of RAM via oral gavage. (6) OVX mice received i.p. injections of 17β-estradiol (E2) (0.1 mg/kg b.w./day) three times per week for 12 weeks.

Results

Micro-CT images showed that oral administration of RAM to OVX mice prevented tibial bone loss, preserved trabecular bone microarchitecture, and improved bone biomechanical properties. RAM administration also showed recovery effects on the levels of TNF-α, OPG and RANKL concentration in OVX-states. Additionally, we found that the mechanism by which RAM elicited anti-osteoporotic effects was by down-regulating the expression of TRAF6 and NFATc1 in RANKL-RANK pathway, a route of osteoclast differentiation, followed by reducing the production of osteoclast differentiation factors, calcitonin receptors and cathepsin K.

Conclusions

Our research strongly suggests that RAM can be clinically used in the prevention and treatment of postmenopausal osteoporosis.     

Achyranthes bidentata root extract prevent OVX-induced osteoporosis in rats

Aim

The objective of the present study was to systematically investigate the effects of Achyranthes bidentata root extract (ABRE) on postmenopausal osteoporosis.

Materials and methods

Eighty 3-month-old female Sprague-Dawley rats were used and randomly assigned into sham-operated group (SHAM) and five ovariectomy (OVX) subgroups, i.e. OVX with vehicle (OVX); OVX with 17β-ethinylestradiol (E₂, 25 μg/kg/day); OVX with ABRE of graded doses (100, 300, or 500 mg/kg/day). Daily oral administration of ABRE or E₂ started on week 4 after OVX for 16 weeks. Bone mass, bone turnover and strength were analyzed by dual-energy X-ray absorptiometry (DEXA), biochemical markers and three-point bending test. The trabecular bone microarchitecture was evaluated by microcomputed tomography (μCT).

Results

16 weeks treatment of ABRE slowed down the body weight gain and prevented the loss of bone mass induced by the OVX. The prevention effect on bone loss was due to altering the rate of bone remodeling, which could be inferred from the decreased level of bone turnover markers, such as serum alkaline phosphatase (ALP), osteocalcin (OC) and urinary deoxypyridinoline (DPD). The changes of urinary calcium and phosphorus excretion provided the same evidence. The treatment could also enhance the bone strength and prevent the deterioration of trabecular microarchitecture.

Conclusions

We conclude that 16 weeks of ABRE treatment improve bone biomechanical quality through modifications of bone mineral density (BMD), and trabecular microarchitecture without hyperplastic effect on uterus, and it might be a potential alternative medicine for treatment of postmenopausal osteoporosis.     

The effects of Labisia pumila var. alata on bone markers and bone calcium in a rat model of post-menopausal osteoporosis

Aim of the study

Postmenopausal osteoporosis is mainly treated with estrogen replacement therapy (ERT). However, ERT causes side effects, mainly breast cancer, uterine cancer and thromboembolic problems. Labisia pumila var. arata (LPva), a herb with phytoestrogenic effects has the potential to be used as an alternative agent to ERT. This study was conducted to determine the effects of LPva on bone biochemical markers and bone calcium content in ovariectomised rats.

Materials and methods

Thirty two Wistar rats were divided into 4 groups, with 8 rats in each group. The first group was sham operated (Sham), the second group was ovariectomised (OVX), the third (LPva) and fourth group (ERT) were also ovariectomised and given LPva 17.5 mg/kg and Premarin^® 64.5 μg/kg, respectively. Blood samples were taken before and after treatment to measure osteocalcin and C-terminal telopeptide of type 1 collagen levels using ELISA while the fifth lumbar bone samples were taken to measure bone calcium content using the Atomic Absorption Spectrophotometer (AAS).

Results

The osteocalcin levels were significantly higher in both the LPva and ERT groups compared to the OVX group. The CTX levels were significantly lower in both the LPva and ERT groups compared to the OVX group. However, only the ERT group had significantly higher bone calcium level compared to the OVX group.

Conclusion

The supplementation of 17.5 mg/kg of LPva to ovariectomised rats for 8 weeks was able to prevent the changes in bone biochemical markers but failed to prevent the bone calcium loss induced by ovariectomy.     

Anti-osteoporosis activity of Cibotium barometz extract on ovariectomy-induced bone loss in rats

Ethnopharmacological relevance

Recent research has confirmed that Cibotium barometz could inhibits osteoclast formation with no affect on BMM cell viability. However, the in?uence of Cibotium barometz on osteoporosis in animals is relatively unknown. The purpose of this study is to systemically investigate the effects of Cibotium barometz extract (CBE) on ovariectomy-induced osteoporosis in rats.

Materials and methods

A total of Seventy-two 3-month-old female Sprague-Dawley rats were used and randomly divided into sham-operated group and five ovariectomized (OVX) groups: OVX with vehicle; OVX with 17β-estradiol (E2, 25 μg/kg/day); OVX with CBE of graded doses (100, 300, or 500 mg/kg/day). Daily oral administration of E2 or CBE began 4 weeks after the surgery and lasted for 16 weeks. Bone mass, bone turnover and strength were analyzed by DEXA, biochemical markers and three-point bending test. The trabecular bone microarchitecture was evaluated by MicroCT.

Results

CBE prevented total BMD decrease in the femur induced by OVX, which was accompanied by a significant decrease in skeletal remodeling, as was evidenced by the decreased levels of the bone turnover markers, such as osteocalcin (OC), alkaline phosphatese (ALP), deoxypyridinoline (DPD), and urinary Ca and P excretions. The treatment could also enhance the bone strength and prevent the deterioration of trabecular microarchitecture.

Conclusions

The present study indicated that Cibotium barometz extract might be a potential alternative medicine for the prevention and treatment of postmenopausal osteoporosis.     

Salvia miltiorrhiza: An ancient Chinese herbal medicine as a source for anti-osteoporotic drugs

Ethnopharmacological relevance

Red sage (Salvia miltiorrhiza Bunge), also known as Danshen in Chinese, has been used historically and is currently exploited in combination with other herbs to treat skeletal diseases in traditional Chinese medicine (TCM). With the advance of modern analytical technology, a multitude of bone-targeting, pharmaceutically active, compounds has been isolated and characterized from various sources of TCM including those produced in Salvia miltiorrhiza root. The aim of the review is to provide a comprehensive overview about the historical TCM interpretation of the action of Salvia miltiorrhiza in osteoporosis, its use clinical trials, its main phytochemical constituents, and its action on bone-resorptive and bone formation-stimulating mechanisms in in vitro and in vivo studies.

Materials and methods

Literature sources used were Pubmed, CNKI.net, Cqvip.com, PubChem, and the Web of Science. For the inquiry, keywords such as Salvia, danshen, osteoporosis, bone, osteoclast and osteoblast were used in various combinations. About 130 research papers and reviews were consulted.

Results

In TCM, the anti-osteopororotic effect of Salvia miltiorrhiza is ascribed to its action on liver and blood stasis as main therapeutic targets defining osteoporosis. 36 clinical trials were identified which used Salvia miltiorrhiza in combination with other herbs and components to treat post-menopausal, senile, and secondary osteoporosis. On average the trials were characterized by high efficacy (>80%) and low toxicity problems. However, various limitations such as small patient samples, short treatment duration, frequent lack of detailed numerical data, and no clear endpoints must be taken into consideration. To date, more than 100 individual compounds have been isolated from this plant and tested in various animal models and biochemical assays. Compounds display anti-resorptive and bone formation-stimulating features targeting different pathways in the bone remodeling cycle. Pathways affected include the activation of osteoblasts, the modulation of osteoclastogenesis, and the inhibition of collagen degradation by cathepsin K.

Conclusions

The inclusion of Salvia miltiorrhiza in more than 30% of all herbal clinical trials successfully targeting osteoporosis has stimulated significant interest in the identification and characterization of individual constituents of this herb. The review highlights the anti-osteoporotic potential of Salvia miltiorrhiza in clinical applications and the potential of the herb to provide potent compounds targeting specific pathways in bone resorption and bone formation.     

Ethanol extract of Peperomia pellucida (Piperaceae) promotes fracture healing by an anabolic effect on osteoblasts

Ethnopharmacological relevance

The whole plant or some part of Peperomia pellucida (L.) HBK is used in some parts of Cameroon as a treatment for fracture healing.

Aim of the study

To evaluate the effect of ethanolic extracts of Peperomia pellucida (L.), a Cameroonian medicinal plant on bone regeneration following bone and marrow injury, and determine the mode of action.

Materials and methods

Ethanol extract of Peperomia pellucida was administered at 100 and 200 mg/kg doses orally to adult female Sprague-Dawley rats having a drill hole injury (0.8 mm) in the femur diaphysis. Vehicle (gum-acacia in distilled water) was given to the control group. After 12 days of treatment, animals were euthanized and femur bones collected. Confocal microscopy of calcein labeling at the drill hole site was performed to evaluate bone regeneration. 3-D microarchitecture of drill hole site was analyzed by micorocomputed tomography. Osteogenic effects of the extract were evaluated by assessing mineralized nodule formation of bone marrow stromal cells and expression of osteogenic genes (mRNA level of type-1 collagen, bone morphogenetic protein-2 and osteocalcin genes) in the femur.

Results

Ethanol extract from Peperomia Pellucida (L.) dose-dependently induced bone regeneration at the fracture site. At 200 mg/kg dose, the extract significantly increased mineral deposition compared to controls. The extract also improved microarchitecture of the regenerating bone evident from increased bone volume fraction, trabecular thickness, trabecular number, and decreased trabecular separation and structure model index. In addition, the extract increased the formation of mineralized nodules from the bone marrow stromal cells. Furthermore, the extract induced the expression of osteogenic genes in the femur including type 1 collagen, osteocalcin and BMP-2, compared to control.

Conclusion

Ethanolic extract of P. pellucid (L.) accelerates fracture repair in rats via stimulatory effects on osteoblast differentiation and mineralization, thereby justifying its traditional use.     

Topical treatment with Tong-Luo-San-Jie gel alleviates bone cancer pain in rats

Ethnopharmacological relevance

The herbal analgesic gel Tong-Luo-San-Jie (TLSJ) and its modifications are used in traditional Chinese medicine to manage cancer pain. However, its mechanisms are still unknown.

Aim of the study

To investigate the effects and mechanisms of TLSJ gel on bone cancer pain in a rat model.

Materials and methods

A bone cancer pain rat model was established by inoculating Walker 256 rat carcinoma cells directly into the right tibial medullary cavity of Sprague–Dawley rats (150–170 g); Phosphate buffered saline (PBS) tibial inoculation was used as control. Cancer-bearing rats were treated twice a day with external TLSJ gel (0.5 g/cm²/day) or inert gel control for 21 day (n=10/group). Behavioral tests such as mechanical threshold and paw withdrawal latency (PWL) were carried out. Osteoclastic activities were determined and carboxyterminal pyridinoline cross-linked type I collagen telopeptides (ICTP) and bone-specific alkaline phosphatase (BAP) concentrations were detected with ELISA after treatment. Adverse effects were monitored, and biochemical and histological tests were performed in naïve rats treated with local TLSJ gel for six weeks.

Results

TLSJ treatment significantly restored bone cancer-induced decrease of PWL and mechanical threshold compared to inert gel. It also decreased the level of blood serum ICTP and BAP and inhibited osteoclast activities. No adverse effects or abnormal biochemical and histological changes were detected after TLSJ treatment.

Conclusion

The present study shows that TLSJ significantly inhibits bone cancer-induced thermal and mechanical sensitization. It suggests that the gel may be useful in managing cancer pain and that it may act by inhibiting osteoclastic activity.     

Antiosteoporotic activity of Davallia formosana

Ethanopharmacological relevance

In Taiwanese folk medicine, Davallia formosana is used to treat bone diseases, including osteoporosis.

Aim

This study evaluated the anti-osteoporotic effect of ethanolic extract derived from Davallia formosana (DFE).

Materials and methods

In this in vitro study, we investigated the inhibitory action of DFE on RANKL-stimulated osteoclastogenesis. The in vivo effects of DFE on bone metabolism were evaluated using ovariectomized (OVX) rats orally administered DFE (200, 500 mg/kg), alendronate (2.5 mg/kg, three times a week) or its vehicle for 12 weeks.

Results

This in vitro study demonstrated that DFE inhibited osteoclast differentiation, and also isolated the active component, (−)-epicatechin 3-O-β-d-allopyranoside (ECAP). DFE did not affect the body or vaginal weight in OVX rats. The bone mineral density and bone calcium content in OVX rats were lower in the control group showing that DFE was able to prevent significant bone loss. In addition, the three point bending test and the microcomputer tomography scanning showed that DFE treatment enhanced bone strength and inhibited the deterioration of trabecular microarchitecture. In the biochemical assay, DFE decreased urinary deoxypyridinoline and calcium concentrations, but did not inhibit serum alkaline phosphatase activities, indicating that it ameliorated bone loss via inhibition of bone reabsorption.

Conclusions

These results suggest that DFE may represent a useful remedy for the treatment of bone reabsorption diseases such as osteoporosis. In addition, ECAP could be used as a marker compound to control the quality of DFE.     

Evaluation of hepatotoxicity and cholestasis in rats treated with EtOH extract of Fructus Psoraleae

Ethnopharmacological relevance

Fructus Psoraleae (FP) has been widely used to heal skin diseases as well as osteoporosis, osteomalacia, and bone fracture. There also exist many clinical reports about FP-induced hepatotoxicity associated with acute cholestatic hepatic injury. However, the FP-induced hepatotoxicity and the underlying mechanisms remain unclear.

Aims of the study

The present study aims to determine the hepatotoxicity of FP in Sprague-Dawley (SD) rats and to investigate the underlying mechanisms.

Materials and methods

Sprague-Dawley rats of both sexes were intragastrically administered with the EtOH extract of FP (EEFP) at doses of 1.875, 1.25 and 0.625 g/kg for 28 day. Body weight, relative liver weight, biochemical analysis, histopathology, the mRNA and protein expression of Cholesterol 7α-hydroxylase (CYP7A1), farnesoid X receptor (FXR), bile-salt export pump (BSEP), multidrug resistance-associated protein 2 (MRP2), multidrug resistance-associated protein 3 (MRP3) were evaluated to study the EEFP-induced hepatotoxicity and its underlying mechanisms.

Results

Many abnormalities were observed in the EEFP-treated groups including suppression of weight gain and food intake, change of some parameters in serum biochemistry, increased weight of liver, and decreased concentration of bile acid in bile. The mRNA and protein expression of CYP7A1, MRP3, MRP2, BSEP increased and the expression of FXR decreased in EEFP-treated female groups; the mRNA and protein of FXR and CYP7A1 decreased and that of the others remained the same in EEFP-treated male groups.

Conclusion

In conclusion, we provide evidence for the first time that EEFP can induce sex-related cholestatic hepatotoxicity, and that female rats are more sensitive to EEFP-induced hepatotoxicity, which involves the destruction of the biosynthesis and transportation of bile acid. Further investigation is still needed to uncover the mechanism of the sex-dimorphic EEFP-induced hepatotoxicity.