Acupuncture Increases Bone Strength by Improving Mass and Structure in Established Osteoporosis after Ovariectomy in Rats

The effects of acupuncture on bone biomechanical properties and histomorphometry in ovariectomized （OVX） rats were studied. Twenty-four 8-week-old female Sprague-Dawley rats were randomly assigned to three groups： sham, model and acupuncture, Rats in the model and acupuncture groups were ovariectomized, while those in the sham group underwent a sham operation. All rats were anesthetized and fastened for 15 minutes, and for the acupuncture group, needling on Pishu （BL20） and Shenshu （BL23） was performed. Blood and urine were collected to measure serum osteocalcin （OC） and urinary calcium, phosphorus or deoxypyridinoline （Dpd）. After 16 weeks of treatment, all the rats were killed and their tibiae and femora were removed. The tibiae were used for analyses of bone histomorphometry and the femora for a three-point bending test. Acupuncture gave significant protection against ovariectomy-caused decline on femoral strength in the mechanical test, increased the trabecular bone volume and thickness, lowered the trabecular separation of tibiae and restricted the excretion of phosphorus and Dpd, while promoting the concentrations of serum osteocalcin as compared with model rats. These results seemed to indicate that acupuncture on the points of Pishu （BL20） and Shenshu （BL23） not only promoted the bone formation but also suppressed the bone resorption induced by OVX in osteoporotic rats, which suggests that it would be a potentially useful and convenient method in preventing osteoporosis.     

Icariin associated with exercise therapy is an effective treatment for postmenopausal osteoporosis

Background  There are no conclusive studies evaluating the interaction between icariin and exercise for treatment of osteoporosis; the efficacy and safety of this treatment combination remains to be evaluated. The purpose of this study was to investigate the effects of icariin treatment combined with exercise therapy on bone parameters and body weight of ovariectomized rats.

Methods  Ovariectomized rats were used as a model of postmenopausal osteoporosis and were exposed to either icariin treatment, exercise, hormone replacement therapy, or a combination of the above. Untreated, ovariectomized rats and sham operated rats were used as controls. After 3 months of experimental interventions the effects of the treatments on the body and uterine weights, the physical and biomechanical properties of bones, and the expression of the osteoblast-specific gene Osterix (Osx), were assessed.

Results  The weight gain of the ovariectomized rats was greater than that of the treated experimental groups. Uterine weight and serum estradiol levels were significantly greater in sham operated and estrogen-treated ovariectomized rats than in the other groups. Biomechanical parameters were improved significantly in ovariectomized rats treated with exercise alone or treated with exercise and icariin. Osx expression was increased in ovariectomized rats treated with exercise and icariin or treated with just icariin.

Conclusions  Exercise combined with icariin had a synergistic effect in the early prevention of osteoporosis in ovariectomized rats. The effects of icariin and exercise on osteoporosis are worth further exploration.

     

The effects of soybean isoflavaones on the mRNA expression of OPG, ODF and M-CSF in bone tissue of ovariectomized rat

Objective： To observe the effects of the soybean isoflavaones on the rnRNA expression of osteoprotegerin（OPG）, osteoclast differentiation factor（ODF） and macrophage colony stimulating factor（M-CSF） in bone tissue of ovariectomized rat, and to investigate the possible pathway to prevent and treat postrmenopausal osteoporosis using soybean isoflavaones. Methods： Thirty healthy adult SD rats were randomly divided into 3 groups： sham-operated group, ovariectomized group and soybean isoflavaones treated group. All rats were ovariectomized except those in sham-operated group. The bone density of the 3-6th lumbar vertebrae was detected after 12 weeks. Total RNA was extracted from femur bone and the mRNA expression of OPG, ODF and M-CSF was examined by real time PCR. Results： Soybean isoflavaones increased the bone density of the lumbar vertebrae in ovariectomized rat and up-regulated the expression of OPG, whereas down-regulated the expression of M-CSF and the ratio of ODF to OPG. Conclusion： The effects of soybean isoflavaones on postmenopausal osteoporosis are tightly correlated with OPG, M-CSF and the ratio of ODF to OPG.     

Humulus lupulus L. Extract Prevents Ovariectomy-Induced Osteoporosis in Mice and Regulates Activities of Osteoblasts and Osteoclasts

Objective:To systematically evaluate the protective effects of Humulus lupulus L.extract(HLE)on osteoporosis mice.Methods:In vivo experiment,a total of 3512-week-old female ICR mice were equally divided into 5 groups:the sham control group(sham);the ovariectomy with vehicle group(OVX);the OVX with estradiol valerate[EV,0.2 mg/(kg·d)];the OVX with low-or high-dose HLE groups[HLE,1 g/(kg·d)and 3 g/(kg·d)],7 in each group.Treatment began 1 week after the ovariectomized surgery and lasted for 12 weeks.Bone mass and trabecular bone mircoarchitecture were evaluated by micro computed tomography,and bone turnover markers in serum were evaluated using enzyme-linked immunosorbent assay(ELISA)kits.In vitro experiment,osteoblasts and osteoclasts were treated with HLE at doses of 0,4,20 and 100μg/mL.Biomarkers for bone formation in osteoblasts and bone resorption in osteoclasts were analyzed.Results:Compared with the OVX group,HLE exerted bone protective effects by the increase of estradiol(P<0.05),the improvement of cancellous bone structure,bone mineral density(P<0.01)and the reduction of serum alkaline phosphatase(ALP),tartrate resistant acid phosphatase(TRAP),bone gla-protein,c-terminal telopeptides of typeⅠcollagen(CTX-Ⅰ)and deoxypyridinoline levels(P<0.01 for all).In vitro experiment,compared with the control group,HLE at 20μg/mL promoted the cell proliferation(P<0.01),and increased the expression of bone morphogenetic protein-2 and osteopontin levels in osteoblasts(both P<0.05).HLE at 100μg/mL increased the osteoblastic ALP activities,and HLE at all dose enhanced the extracellular matrix mineralization(both P<0.01).Furthermore,compared with the control group,HLE at 20μg/m L and 100μg/m L inhibited osteoclastic TRAP activity(P<0.01),and reduced the expression of matrix metalloproteinase-9 and cathepsin K(both P<0.05).Conclusion:HLE may protect against bone loss,and have potentials in the treatment of osteoporosis.     

Effect of Epimedium-derived Phytoestrogen on Bone Turnover and Bone Microarchitecture in OVX-induced Osteoporotic Rats

To investigate the preventive effect of epimedium-derived phytoestrogen(PE) on osteo-porosis induced by ovariectomy(OVX) in rats,11-month-old female Wistar rats were randomly di-vided into Sham,OVX and PE groups.One week after OVX,daily oral administration of PE(0.4 g·kg-1·day-1) started in PE group,and rats in Sham and OVX groups were given vehicle accordingly.The administrations lasted for 12 weeks.The biological markers including serum osteocalcin(OC) and urinary deoxypyridinoline(DPD) for bone turnover were evaluated at the end of the 12th week.On the 13th week,all the rats were sacrificed.The right proximal tibiae were removed,subjected to micro CT for determination of trabecular bone structure and then bone histomorphometry was per-formed to assess bone remodeling.The OVX rats were in a high bone turnover status as evidenced by increased bone formation markers and bone resorption markers.Treatment with PE could suppress the high bone turnover rate in OVX rats.Micro CT data revealed that PE treatment could ameliorate the deterioration of the micro-architecture of proximal tibiae induced by OVX,as demonstrated by greater bone volume,increased trabecular thickness and less trabecular separation in PE group in comparison with OVX group.The static and dynamic parameters of bone histomorphometry indi-cated that there were significant increases in bone formation variables and significant decreases in bone resorption variables between PE and OVX groups.The findings suggest that PE has a beneficial effect on trabecular bone in OVX rat model and this effect is possibly associated with stimulation of bone formation as well as inhibition of bone resorption.     

Positive Effects of Qing'e Pill(青娥丸)on Trabecular Microarchitecture and its Mechanical Properties in Osteopenic Ovariectomised Mice

Objective: To investigate the impact of Qing'e Pill(青娥丸,QEP) on the cancellous bone microstructure and its effect on the level of β-catenin in a mouse model of postmenopausal osteoporosis.Methods: Ninety-six 8-week-old speci?c pathogen free C57 BL/6 mice were randomly divided into 4 groups(24/group): sham,ovariectomised osteoporosis model,oestradiol-treated,and QEP-treated groups.Three months after surgery,the third lumbar vertebra and left femur of the animals were dissected and scanned using micro-computed tomography(micro-CT) to acquire three-dimensional(3 D) parameters of their cancellous bone microstructure.The impact of ovariectomy,the effect of oestradiol and QEP intervention on cancellous bone microstructure,and the expression of β-catenin were evaluated.Results: The oestradioland the QEP-treated groups exhibited a signi?cant increase in the bone volume fraction,trabecular number,trabecular thickness,bone surface to bone volume ratio(BS/BV),and β-catenin expression compared with those of the model group(P0.05).In contrast,the structure model index,trabecular separation,and BS/BV were signi?cantly decreased compared with those of the ovariectomised osteoporosis model group(P0.05).No differences were observed in the above parameters between animals of the QEP-and oestradiol-treated groups.Conclusions: The increased β-catenin expression may be the mechanism underlying QEP's improvement of the cancellous bone microstructure in ovariectomised mice.Our ?ndings provide a scienti?c rationale for using QEP as a dietary supplement to prevent bone loss in postmenopausal women.     

Ursolic Acid Prevents Retinoic Acid-Induced Bone Loss in Rats

Objective: To examine the effects of ursolic acid(UA) on mitigating retinoic acid(RA)-induced osteoporosis in rats. Methods: Fifty female Sprague-Dawley rats were randomly divided into the control group(n=10) and the osteoporosis group(n=40). The 40 osteoporosis rats were induced by 75 mg/(kg·d) RA once daily for 2 weeks, and then were randomly assigned to vehicle control(model), low-, middle-, and high-dose UA [(UA-L, UA-M, UA-H; 30, 60, 120 mg/(kg·d), respectively] groups(10 rats each). UA were administered once daily to the rats from the 3 rd weeks for up to 4 weeks by gavage. Bone turnover markers [serum alkaline phosphatase(ALP), osteocalcin(OCN), urine deoxypyridinoline(DPD)] and other parameters, including serum calcium(S-Ca), serum phosphorus(S-P), urine calcium(U-Ca), urine phosphorus(U-P), and bone mineral density(BMD) of the femur, 4 th lumbar vertebra and tibia, bone biomechanical properties and trabecular microarchitecture, were measured. Results: The osteoporosis in rats was successfully induced by RA. Compared with the model group, UA-M and UA-H significantly reversed the RA-induced changes in S-P, U-Ca, U-P, ALP, OCN and urine DPD ratio and markedly enhanced the BMD of right femur, 4 th lumbar vertebra and tibia(P0.05 or P0.01). Further, biomechanical test and microcomputed tomography evaluation also showed that UA-H drastically improved biomechanical properties and trabecular microarchitecture(P0.05 or P0.01). Conclusion: UA could promote bone formation, increase osteoblastic activity and reduce osteoclastic activity in rats, indicating that UA might be a potential therapeutic of RA-induced acute osteoporosis.     

A Trial Study of Propranolol and Zhigancao Decoction on the Central Depressant and Anti-osteoporosic Action in Ovariectomized Rats

Objective: This study was originally designed to observe the effects of propranolol (a β-blocker) and Zhigancao Decoction (炙甘草汤ZGCD) on bone mass in ovariectomized rats. Methods: Thirty-eight female Sprague-Dawley rats were divided into four groups initially, a sham-operated group (Sham, n=7), a model ovariectomized (OVX) group (Model, n=7), a propranolol group (Pro, n=12) and a ZGCD group (ZGCD, n=12). After 15 weeks of treatment, the expected effects were not found. In order to verify the situations of the experiment, we modified the study by administering calcitonin to a subgroup of the tested Pro and ZGCD rats. Results: The Pro and ZGCD treatments showed decreased heart rate and plasma norepinephrine level, but neither an increased bone mass nor any bone metabolism differences from the model rats were found. However, the OVX-induced bone loss was prevented by the sequent treatment of calcitonin. Conclusions: The results provide no evidence that the β-blocker propranolol may stimulate bone formation, and do not justify its use for clinical treatment of osteoporosis.     

Abortifacient Potential of Aqueous Extract of Senna alata Leaves in Rats

Objective To study the abortifacient activity of aqueous extract of Senna alata leaves in female Wistar rats.Methods Pregnant rats weighing 143.65 ± 13.11 g were completely randomized into 5 groups（A-E）.Rats in group A（negative control） were orally administered,once daily with 0.5 ml of distilled water on days 10-18 post-coitum while those in groups B,C,D,and E were treated exactly like the negative control except they received the same volume containing 2.85 mg/kg body weight of mifepristone（group B,reference drug）,250,500 and 1 000 mg/kg body weight of the extract respectively（groups C-E）.Results Phytochemical screening of the extract showed positive results for saponins（1.22%）,flavonoids（1.06%）,cardiac glycosides（0.20%）,cardenolides and dienolides（0.18%）,phenolics（0.44%） and alkaloids（0.52%）.Respiratory distress,salivation,diarrhoea,changes in the appearance of hair as well as maternal mortality were not observed at any time during the exposure period except in the mifepristone-treated animals where there was episode of diarrhoea and tiredness.The extract significantly reduced（P〈0.05） the number of life foetus,weight and survival ratio of the foetus,numbers of implantations and Corpora lutea,implantation index,progesterone,prolactin,estradiol,follicle stimulating and luteinizing hormones whereas the number of dead foetus,number and percentage of rats that aborted,percentage vaginal opening,resorption index,pre-and post-implantation losses increased significantly.In addition,mifepristone-treated animals produced resorption index that compared well with the distilled water control.There was also no dead or life foetus and serum progesterone concentration was increased in the animals treated with mifepristone.All cases of abortion were accompanied with vaginal bleeding.Although,the final weight of the rats increased significantly,the feed and water intake were not significantly altered in all the treatment groups.The weight of the uterus,uterine-body weight ratio,length of the right uterus horn and uterine cholesterol decreased significantly in all the treatment groups.The uterine alkaline phosphatase activity and glucose concentration increased in only the extract-treated animals whereas mifepristone decreased the uterine alkaline phosphatase activity and glucose content of the animals.Conclusion This study has provided evidence to the age-long claim of S.alata leaves in ＂washing the uterus＂.The abortifacient properties were most pronounced at 500 and 1 000 mg/kg body weight of the extract and were similar to the animals treated with 2.85 mg/kg body weight of mifepristone.Hormonal influence,changes in implantation site,estrogenicity and uterogenicity are suggested as possible mechanism of abortifacient activity of aqueous extract of S.alata leaves.Overall,the extract may be used as an abortifacient especially at 500 and 1 000 mg/kg body weight and therefore not safe for consumption as oral remedy during pregnancy.     

EFFECTS OF XW630 ON BONE FORMATION IN OVARIECTOMIZED RATS AND IN HUMAN OSTEOBLAST-LIKE CELLS

Objective. To study the effects of XW630 on bone formation in overiectomized(OVX) rats and in hu-man osteoblast-like cell line TE85. Method. Bone histomorphometric analysis was performed with undecalcified bone sections and tetracy-cline intraperitoneally labeling. Results. Compared with that of OVX rats, the static data of trabecular bone volume(TBV)/total tis-sue volume(TTV), TBV/sponge bone volume(SBV) and mean trabecular plate density (MTPD) were en-hanced while mean trabecular plate spacing(MTPS) decreased after treated with XW630 for 13w. The dy-namic data of single-labeled surface [Sfract(s)], double-labeled surface[Sfract(d)],Sfract(d 1/2s),tra-becular osteoid surface(TOS), and bone formation rate in tissue level (Svf) were increased and osteoid maturation period (OMP) shortened in XW630 group. In osteoblast-like cells, both ^3H-thymidine incorpo-ration and cell count increased after treated with XW630 for 48. Treated with XW630 for 12～ 18h,in-ducible nitric oxide synthase(iNOS) activity and cGMP content increased in time-dependent manners. Conclusions. XW630 enhanced bone activation frequency and increased trabecular connectivity, stabili-ty, and strength. The cellular mechanism related to effects of XW630 on bone formation in ovariectomized     

Experimental research into using bone-strengthening pill to treat ischemic necrosis of femoral head

OBJECTIVE:To study the use of a bone-strengthening pill to treat ischemic necrosis of the femoral head.METHODS:A model of castration in rats with osteoporosis and a model of ischemic necrosis in the bilateral femoral heads of young rats were used to detect bone density,bone formation measurements,lipid deposition in the femoral head,the distribution of capillaries in the femoral head and other relevant biochemical indices.RESULTS:In model rats,bone-strengthening pills were able to increase the bone density,the unit volume of bony trabeculae,the width of bony trabeculae,and the unit volume of the bony cortex.They could also enhance the mineral apposition rate in the femoral head and the seam width of osteoid deposition.Furthermore,there was a reduction in lipid deposition in the femoral head,and enriched distribution of arteries in the femoral head.Bone-related biochemical detections showed an increase in alkaline phosphatase staining and a decrease in tartaric acidic phosphatase staining.CONCLUSION:The results show that bone-strengthening pills can obviously promote bony growth,cause an increase in bone density,restore blood circulation,inhibit the occurrence and development of bony necrosis,and accelerate the repair of necrotic bones,with no toxic side-effects.     

Preventive Effects of Nitroglycerine on Glucocorticoid-induced Osteoporosis in Growing Rats

The preventive effects of nitroglycerine （NG） on glucocorticoid-induced osteoporosis in growing rats were studied. Three-month-old female Wistar rats were randomly divided into control group （CON）, dexamethasone group （DXM）, DXM plus a low dose NG group （NG-L）, DXM plus a middle dose NG group （NG-M） and DXM plus a high dose NG group （NG-H）, 8 rats in each group. The rat model of osteoporosis was developed by intramuscular injection of dexamethasone twice a week. NG 0.2, 0.4 and 1.0 mg/kg was administered by oral gavages to the treatment groups every day for 12 weeks. Rats in CON group and DXM group were treated with normal saline of the same amount. After the treatment, the bone mineral density （BMD） and bone metabolism-associated biochemical markers were determined. Compared with CON group, BMD of lumbar spine and femur in DXM group was decreased significantly （P〈0.05 and P〈0.01 respectively）, blood BGP levels and NO levels reduced （both P〈0.01）, and TRAP level increased （P〈0.05）. As compared with DXM group, BMD, serum BGP and NO were increased, and TRAP decreased in NG-L group and NG-M group, but had no significant difference in comparison to CON group. All the markers other than serum NO and TRAP levels had no significant difference between NG-H group and DXM group. It was concluded that low or middle doses of NG could prevent glucocorticoid-induced bone loss in growing rats, but high dose of NG could not. Supplement with NO donor could be considered as a preventive treatment for glucocorticoid-induced osteoporosis in a developing skeleton.     

Effect of Wuziyanzong Pill on sperm quality and calcium ion content in oligoasthenospermia rats

OBJECTIVE:To study the effect of Wuziyanzong treatment on the sperm quality and content of calcium ions(Ca 2+) in oligoasthenospermia rats.METHODS:A model of oligoasthenospermia was induced in 50 Sprague Dawley rats by treatment with tripterygium glycosides at 30 mg/kg per day for 8 weeks.They were divided randomly into a model group,a positive group(Huangjingzanyu capsule,3.01 g/kg),and low,medium and high dose Wuziyanzong treatment groups(2.30,4.60,9.20 g/kg crude drug respectively) with 10 in each group.Another 10 rats were used as a control group.The rats in the control and model groups were administered distilled water,while the rats in the remaining groups were administered Wuziyanzong for 30 d.The epididymides were removed,spermatozoa recovered and the sperm density and viability were measured.The spermatozoa were purified and the contents of Ca 2+ in the cytoplasm and mitochondria were detected by flow cytometry and atomic absorption spectrometry,respectively.RESULTS:After 8 weeks of treatment with tripterygium glycosides,the sperm density,sperm activity and the Ca 2 + content of spermatozoa in the model rats were all significantly decreased compared with the control group(all P<0.05).After 30 d treatment,the sperm density and activity improved and the Ca 2 + content of sperm were increased significantly in the medium and high dose Wuziyanzong treatment groups in comparison with the model group(all P<0.05).CONCLUSION:The Wuziyanzong treatment increased sperm density,improved sperm viability and enhanced the content of Ca 2+ in the sperm cytoplasm and mitochondria in this rat model of oligoasthenospermia.     

Effects of Xuesetong Soft Capsules(血塞通软胶囊) on angiogenesis and VEGF mRNA expression in ischemic myocardium in rats with myocardial infarction

OBJECTIVE:To observe the effects of Xuesetong Soft Capsules(血塞通软胶囊,Notoginseng total saponin) on angiogenesis and vascular endothelial growth factor(VEGF) mRNA expression in ischemic myocardium of rats with myocardial infarction.METHODS:The left coronary artery of rats was ligated to establish the animal model of acute myocardial infarction.Rats were randomly divided into Xuesetong Soft Capsule,Shexiangbaoxin Pill(positive control),model(negative control) and sham operation groups.After 6 weeks,microvessel count(MVC),microvessel density(MVD) and VEGF mRNA expressioninischemicmyoc ardium were evaluated.RESULTS:MVC and MVD in the myocardial infarct border area in model,Shexiangbaoxin Pill and Xuesetong Soft Capsule groups significantly increased compared with those of the sham operation group(P<0.05).MVC and MVD in the myocardial infarct border area in Xuesetong Soft Capsule and Shexiangbaoxin Pill groups significantly increased compared with those of the model group(P<0.05).No significant differences between Xuesetong Soft Capsule and Shexiangbaoxin Pill groups were observed(P>0.05).The model group showed signifi-cantly higher VEGF mRNA expression than that in the sham operation group(P<0.05).Xuesetong Soft Capsule and Shexiangbaoxin Pill groups showed significantly higher VEGF mRNA expression than that of the model group(P<0.05).No significant difference between Xuesetong Soft Capsule and the Shexiangbaoxin Pill groups was observed(P>0.05).CONCLUSION:Xuesetong Soft Capsules promote angiogenesis in ischemic myocardium after myocardial infarction and the mechanism may be associated withVEGF mRNA expression.     

Edible Bird's Nest Attenuates Menopause-Related Bone Degeneration in Rats via Increasing Bone Estrogen-Receptor Expression

Objective:To investigate the mechanistic basis for the attenuation of bone degeneration by edible bird's nest(EBN)in ovariectomized rats.Methods:Forty-two female Sprage-Dawley rats were randomized into 7 groups(6 in each group).The ovariectomized(OVX)and OVX+6%,3%,and 1.5%EBN and OVX+estrogen groups were given standard rat chow alone,standard rat chow+6%,3%,and 1.5%EBN,or standard rat chow+estrogen therapy(0.2mg/kg per day),respectively.The sham-operation group was surgically opened without removing the ovaries.The control group did not have any surgical intervention.After 12 weeks of intervention,blood samples were taken for serum estrogen,osteocalcin,and osteoprotegerin,as well as the measurement of magnesium,calcium abd zinc concentrations.While femurs were removed from the surrounding muscles to measure bone mass density using the X-ray edge detection technique,then collected for histology and estrogen receptor(ER)immunohistochemistry.Results:Ovariectomy altered serum estrogen levels resulting in increased food intake and weight gain,while estrogen and EBN supplementation attenuated these changes.Ovariectomy also reduced bone ER expression and density,and the production of osteopcalcin and osteorotegerin,which are important pro-osteoplastic hormones that promote bone mineraliztion and density.Conversely,estrogen and EBN increased serum estrogen levels leading to increased bone ER expression,pro-osteoplastic hormone production and bone density(all P<0.05).Conclusion:EBN could be used as a safe alternative to hormone replacement therapys for managing menopausal complications like bone degeneration.     

阿司匹林治疗大鼠绝经后骨质疏松的实验研究

目的 探讨阿司匹林对大鼠绝经后骨质疏松的治疗作用及其相关机制.方法 将40只3月龄SD雌性大鼠随机分为5组:对照组(Sham)、去势组(OVX)及3个阿司匹林治疗组(Aspirin),每组8只.OVX及Aspirin组采用卵巢切除法去势,饲养3个月建立骨质疏松模型.确认模型建立成功后予灌胃治疗,阿司匹林治疗组剂量分别为8.93、26.79、80.36 mg/kg(A1、A2及A3),每天灌胃1次.OVX及Sham组每天予等量生理盐水灌胃.术前、术后3个月及灌胃3个月时用双能X线吸收骨密度测量仪(DXA)测量腰椎骨密度;灌胃3个月时测量外围血碱性磷酸酶及骨钙素,采用Micro-CT对骨小梁微观三维形态结构进行分析,组织病理学对骨小梁微观结构进行观察,用生物力学实验评估椎体及股骨干的力学强度.结果 DXA分析显示:去势3个月后,大鼠骨密度较术前及对照组明显降低,骨质疏松模型成功建立.灌胃治疗3个月后,阿司匹林治疗组椎体的BMD明显高于OVX组.外围血检测显示:阿司匹林治疗组的碱性磷酸酶明显低于OVX组,而骨钙素与OVX组无明显差别.组织形态学结果显示,阿司匹林治疗组骨小梁的连接、粗细较OVX组好.Micro-CT分析表明:阿司匹林治疗组腰椎椎体骨小梁厚度、骨小梁数量及骨密度率均显著高于OVX组,并且A3组与Sham组相近.生物力学结果示,阿司匹林治疗组腰椎压缩载荷及股骨干三点弯曲载荷均明显高于OVX组.结论 阿司匹林能促进大鼠绝经后骨质疏松骨小梁的改建,改善骨小梁的三维结构,增加骨质的骨密度和力学强度,有可能成为治疗骨质疏松的新型药物之一.

Abstract：

Objective To explore the therapeutic effects of aspirin on postmenopausal osteoporosis and understand its action mechanism. Methods Forty three-month-old female SD rats were randomly divided into 5 groups ( n = 8 each): sham group, OVX group and aspirin groups ( A1, A2 & A3 ). The OVX and aspirin groups were ovariectomized (OVX). All rats underwent BMD (bone mineral density) scan at the time of OVX and 3 months after OVX. After the animal model of osteoporosis was established, aspirin groups were intragastrically administered at a dose of 8.93 mg· kg-1 · d-1 (A1), 26. 79 mg · kg-1 · d-1(A2) and 80.36 mg · kg-1 · d-1 (A3) daily in OVX rats. Three months later, the BMD and microarchitecture of vertebrae were measured by dual-energy X-ray absorptiometry and microtomograph. Alkaline phosphatase and osteocalcin were measured in peripheral blood. The trabecular architecture changes were observed by histomorphology. Axial compression tests were used to evaluate the mechanical properties of vertebral specimens and three-point bending tests used for femur shaft. Results Three months after ovariectomy, BMD was significantly lower than preoperative. BMD in aspirin treated groups was significantly higher than that in OVX group. Alkaline phosphatase in peripheral blood decreased significantly in aspirin groups than those in OVX, but osteocalcin had no significant difference between aspirin and OVX groups.The microtomograph reconstruction analysis also showed that the trabecular thickness, trabecular number and BMD increased significantly in aspirin groups than those in OVX. And there was no significant difference between A3 and sham groups. The results of biomechanical test showed that the maximum compression load of lumbar spine and three-point bending load of femur shaft were significantly higher in aspirin groups than those in OVX group. Conclusion Aspirin can promote trabecular bone remodeling, improve threedimensional structure of trabecular bone and increase bone density of cancellous in osteoporotic rats by stimulating bone formation. It may become a new drug for the treatment of osteoporosis.     

补肾滋阴中药对去卵巢大鼠糖尿病骨质疏松模型皮质骨宏观几何结构及生物力学的影响

目的 探讨补肾滋阴中药对去卵巢大鼠糖尿病骨质疏松模型宏观几何结构及生物力学的影响.方法 所有大鼠随机分为正常对照组、模型对照组和中药治疗组、乙烯雌酚组,每组各10只.术后90 d开始给药治疗,术后24周处死后分别测定左股骨中段截面宏观几何参数及右股骨三点弯曲实验.结果 模型对照组宏观几何参数与其他组相比,髓腔明显扩大,骨皮质变薄(P＜0.05),补肾滋阴中药及乙烯雌酚组虽然髓腔无缩小但骨皮质明显增厚.生物力学检查结果显示模型对照组及正常对照组最大应力、最大应变低于中药治疗组及乙烯雌酚组(P＜0.05).结论 补肾滋阴中药对糖尿病骨质疏松有明显的治疗作用,可以明显改善糖尿病大鼠骨质疏松模型的骨质量,提高骨的生物力学特性. Abstract： Objective To investigate the influence of bushenziyin recipe on macroscopic geometry structure and biomechanical properties of cortical bone of femur in ovariectomized osteoporosis diabetic rat models.Methods Forty femina rats at the age of 12 months old were selected and divided randomly into four groups( normal control group, experimental control group, bushenziyin recipe group and diethylstilbestrol group).First, normal control group were operated shamly and injected streptozotocin, and the other groups were established successfully the model of rats with diabetic osteoporosis by the compound method of ovariectomy plus streptozotocin.The time point of administrating rats drugs was at 90 days after operation,the time point of sacrificing the rats was at 24 weeks after operation.Macroscopic geometry structure were measured and three -point bending test were done after killing the rats.Results The diameter of medullary canal were increased in the experimental group compared with other groups ( P ＜ 0.05 ); in bushenziyin recipe group and diethylstilbestrol group, the diameter of medullary canal were not decreased sigficantly (P ＞ 0.05 ), the thickness of cortical bone were all increased( P ＜ 0.05 ).Biomechanical properties show that in normal control group and experimental control group maximum stress and maximum straining were decreased versus bushenziyin recipe group and diethylstilbestrol group ( P ＜ 0.05 ).Conclusions Bushenziyin recipe can raise the quality of cortical bone of femur in the ovariectomized osteoporosis diabetic rat models,busehenziyin recipe can improve biomechanical properties of cortical bone.     

Effects of Zuogui Pill (左归丸) on Wnt Singal Transduction in Rats with Glucocorticoid-induced Osteoporosis

Objective: To reveal the mechanism of Zuogui Pill (左归丸) in treatment of glucocorticoid-induced osteoporosis from the angle of the Wnt signal transduction pathway and to provide further experimental evidence for expounding the scientific connotation of "the kidney dominating the bones" in TCM. Methods: Forty-two male Wistar rats were selected and randomly divided into three groups, control group (n=12), model group (n=15) and Zuogui Pill group (n=15). Form the beginning, The rats were injected dexamethasone for eight weeks to make the model of osteoporosis, and the Zuogui Pill were administered intragastrically to the rats of Zuogui Pill group for eight weeks. The relative morphological parameters were measured in the undecalcified tibial slices. And the protein expression levels of Wnt1, LRP-5 and β-catenin in rat tibial osteoblasts (OB) and bone marrow stromal cells (BMC) were detected by immunohistochemistry. Results: Compared with the control group, TBV% and TFS% decreased significantly, while TRS% increased significantly, and the protein expression of Wnt1, LRP-5 and β-catenin in OB and BMC decreased significantly in the model group. And compared with the model group, TBV% and TFS% increased significantly, and expression levels of Wnt1, LRP-5 and β-catenin proteins increased significantly in the Zuogui pill group. Conclusion: Zuogui Pill can prevent and treat glucocorticoid-induced osteoporosis in rats by up-regulating the expression of the key signal molecules Wnt1, LRP-5 and β-catenin in Wnt signal transduction pathway.