Relationship Of Osteoporosis And Nutrition Of Diet For Aged And Effect Of Intervention Empirical Study Of Flavonoids

Osteoporosis is a frequent chronic senile disease, and in which bones become fragile and more easily to break. Osteoporosis is not any clinical symptom in the initial stage. If not prevented or if left untreated, osteoporosis can produce and progress stealthily until a bone breaks. These fractures, occur typically in the hip, spine, and wrist. Any bone can be affected, but of special concern are fractures of the hip and spine. A hip bone fracture almost always requires hospitalization and major surgery. Spinal or vertebral fractures also have serious consequences, including angular curvature, severe back pain, and deformity. It can impair a person's ability to walk unassisted and may cause prolonged or permanent disability or even death.The osteoporosis animal model was established by many scholars with castrated method. The sex hormone level of the model animal was lower, the bone mineral content was more descend also in the research. The research of the Cheng SL discovers that the soybean isoflavone has estrogenic function, it can valid decrease bone lose and increase the bone formation. Arjmandi definitely hints the soybean isoflavone can contain the bone reservation function, point out that consume soybean or its isoflavones to contribute the bone substance's hold out for long time, over a long period of time. But the mechanism of the isoflavone anti osteoporosis is still without thorough research. In this study we investigate the relationship of diet for aged and osteoporosis and research the intervention effect of soy isoflavone and celosiae cristatae flavone.Part I Relationship of nutrition and bone mineral density in Elderly People1. Relationship between BMD and Zn, Cu, Ca Levels in the Hair and Meal in Elderly PeopleSummary: The relationship between bone mineral density (BMD) and Zn, Cu, Ca levels in the meal and hair of urban and rural elderly people were studied. 470 subjects above 60 years old (urban 205 and rural 265), 178 males with an average age of 65.70 ± 3.48y and 292 females with an average age of 65.90±4.02y, were inquired. The BMD and Zn, Cu, Ca levels in the meal and hair were measured. The detected BMD in urban and rural female old people was significantly lower than that of the males; The Ca, Zn contents in the meal of the urban females were significantly lower than those of the urban males; The Ca, Zn in the meal and Zn in the hair of the rural females were significantly lower than those of rural males (P< 0.05 or 0.01). The BMD, Ca intakes, Ca and Zn in the hair of the rural old people were significantly lower than those of the urban old people (P< 0.05 or 0.01). There was a correlation between BMD with the Ca, Zn of the hair and dietary Ca, Zn, Cu or between dietary Zn with Ca, Zn in the hair and Ca, Cu intakes. The Zn, Cu and Ca levels in the meal nutrients were correlated with BMD to some degrees. Lack of Ca and Zn in the meal can cause the reduction of BMD. 2. P Dag Analysis of Effects of daily nutrition intake on BMD Objectives To study the effects of daily mineral intake on bone mineral density (BMD) of old people.Data Measures The daily nutrition intake and health were investigated in 309 old people aged over 60 years. 132 male, aged 65.4±4.0y, 177 female aged 66.7±5.8y.Based on the data, the regression analysis and causal analysis are presented and thus a partial directed acyclic graph (p dag) are plotted to demonstrate a definite causal relation. Results The paper shows that BMD in female were lower than that in male. For healthy people, the daily intake of Calcium, protein, Vitamin C of male was higher than that of health female. The daily intake of Calcium, protein, Vitamin C of osteoporosis case was lower than that of health people of the same sex. BMD correlation with the daily intake of Calcium, protein, Vitamin C is significantly positive, which is consistent with directed acyclic graph (p dag) analysis.Conclude It was suggest that there is definite relation between the daily mineral intake and BMD. The lack of daily intake of Calcium, protein, Vitamin C may decrease BMD. Part II The effect of soy isoflavonord and flavonoid of celosia cristataon increment, mineralization and partial related gene expression in vitro cultured1. Effect of flavonoid of celosia cristata on mineralization and IGF-1 expression invitro cultured osteoblastsObjective To study the effect of flavonoid of celosia cristata extract on mineralization and IGF-1 expression of cultured osteoblasts from ratMethods The osteoblastic cells from cranial bone of newborn rat was culturedthe. The effect of mineralization was detected by alizarin bordeaux stain(ARS). The effect of flavonoid of celosia cristata extract on IGF-1 expression of cultured osteoblasts was detected by SP method. The expression level was monitored with HPIAS1000 photograph analysismeter. Results The IGF-1 of third cultured osteoblasts cultured used flavonoid of celosia cristataextract was positive expression .The IGF-1 positive expression of flavonoid of celosiacristata extract group was higher significantly than negative control group (p<0.05) .Conclusion The flavonoid of celosia cristata extract may increase the mineralization andIGF-1 positive expression of cultured osteoblasts from rat2. Effect of soybean isoflavonet on TGF-β1, TGF-βR1 and TGF-βR2 expression of cultured osteoblasts from ratObjective To study the effect of soybean isoflavone on TGF-β1 , , TGF-βR1 andTGF-βR2 expression of cultured osteoblasts from rat.Methods The osteoblstic cells from cranial bone of newborn rat was cultured. The SPmethod was used to detect the effect of soybean isoflavone on TGF-β1, TGF-βR1 andTGF-βR2 expression of cultured osteoblasts.The expression level was monitored withHPIAS₁000 photograph analysismeter.Results The TGF-β1, TGF-βR1 and TGF-βR2 of osteoblasts cultured used soybeanisoflavone was positive expression .The TGF-β₁ , TGF-βR1 and TGF-βR2 positiveexpression of soybean isoflavone group was higher significantly than negative controlgroup (p<0.05) ..Conclusion The soybean isoflavone may increase the TGF-β₁ , TGF-βR1 and TGF-βR2positive expression of cultured osteoblasts from rat.3. Effects of soybean isoflavone and calcium on proliferation, differentiation andmineralization of cultured osteoblasts in vitroObjective To explore the effects of soybean isoflavone and calcium on proliferation , differentiation and mineralization of cultured osteoblastic cells.Methods Osteoblastic cells from newborn rat calvarial was cultured. Osteoblastic cells proliferation was masured by MTT, activity of ALP was observed with Golden's method and ARS was used to measure the mineral nodes of osteoblasts. Results The calcium group had not diferent frome the control group on stimulating the proliferation, differentiation and mineralization of rat calvarial osteoblastic cells.The soybean isoflavone group had significant effcts on stimulating the proliferation , differentiation of osteoblasts and increase the forming of the mineral nodes of osteoblasts in rat. calcium and soybean isoflavone group had more benefit for forming mineral nodes. Conclusion The soybean isoflavone stimulates the proliferation, differentiation and mineralization of rat calvarial osteoblastic cells, and increase calcium in the same time is more benefit for mineralization. 4. Effect of soybean isoflavonet on bone morphogenetic protein 2 expression ofcultured osteoblasts from ratObjective To study the effect of soybean isoflavone on bone morphogenetic protein 2 (BMP₂)expression of cultured osteoblasts from rat.Methods The osteoblstic cells from cranial bone of newborn rat was cultured. Cultured rat osteoblasts were treated with soybean isoflavone in difierent concentrations. Total RNA was extracted the from the cells. BMP₂ a were amplified by RT—PCR. β -actin cDNA was also amplified at the same time and used as an internal standard. The SP method was used to detect the effect of soybean isoflavone on BMP₂ expression of cultured osteoblasts. The expression level was monitored with HPIAS₂000 photograph analysismeter. The pictures of PCR production was scanned. The specific values of density of BMP₂/β -actin were calculated, and the expression levels of BMP₂ were determined.Results Soybean isoflavone increased the level of BMP₂ in rat osteoblasts and increased the expressions of BMP₂ genes at the level of transcripaon and revealed dose-effect relation, (p<0.01) .Conclusion The soybean isoflavone induces the proliferation and differentiated of osteoblast probably by increasing the expressions of BMP₂ genes. Part III The effect of soy isoflavonord and flavonoid of celosia cristataon bone metabolism index in disuse osteoporosis and ovariectomized rat1. The effect of calcium and extraction of cristata flavonoid on disuse osteoporosis in ratsObjective To research the effect of calcium and extraction of cristata flavonoid on disuse osteoporosis in rats.Method 40 healthy , male SD rats were selected, and randomly divided into 5 groups. The group 1 (normal group) and group 2(positive group) were given with distilled water by intragastrition. Group 2-5 rats were created to the model of disuse osteoporosis. Group 3, 4 and 5 were difference given with cristata flavonoid, calcium carbonate, cristata flavonoid with calcium by intragastrition. After 8 weeks experiment, the bone mineral density(BMD), femur computer tomography (CT), serum malondialdehyde (MDA), superoxide dismutase (SOD), and urine calcium, urine hydroxyproline (HOP), urine creatinine were detected.Results It was showed that the femur weight, ash weight , CT value in positive group were significantly lower than that of normal group and cristata flavonoid group(P<0.05 or P<0.01). Compare whit positive groups , urinary calcium and HOP were lower and serum calcium, femur BMD, serum SOD were higher in cristata flavonoid and cristata flavonoid with calcium groups(P<0.05 or P<0.01).Conclusion It was suggested that calcium and cristata flavonoid can effect the bone matrix metabolism of disuse osteoporosis in rats, cristata flavonoid with calcium had more effect on disuse osteoporosis rats. 2. Effect of cristata L flavonoid on prevetive osteoporosis in ovariectomized rats Objective To study the effect of cristata L flavonoid on urine mineral and the expression of bone BMP₂ , and phagocytosis function of Mononuclear macrophage in ovariectomized rats.Methods Twenty one female SD rats were randomly divided into three groups: sham, ovariectomized(OVX), ovariectomized with cristata L flavonoid (CRIST, 100mg/kg^-· d). The content of urinary calcium, sodium, potassium were measured after ten weeks. The Streptavidin-Peroxidase Histostain^TM-Plus Kits was used to detect the expression of BMP₂ in bone. And phagocytosis test of mononuclear macrophage in vitro, carbon clearance measurement were carried out to determine the phagocytotic functions of mononuclear macrophage.Results Compared with sham group the calcium, sodium in urine expression of ovariectomized group were higher and the potassium of urine and the expression of BMP₂ , and the phago- percentage, and phagocytic index and the carbon clearance index were significantly decreased in OVX group(P<0.05 or <0.01). Compare with OVX group the urine calcium, sodium expression, and the phago- percentage, carbon phagocytic index were significantly in CRIST group, but the bone BMP₂ expression was significantly higher than that of OVX group (P<0.05 or<0.01 ) . Conclusion The cristat L flavonoid could decrease the urinary calcium loss, and increased the expression of BMP₂ , and increased the phagocytotic functions of mononuclear macrophage. It may be advantageous to prevent the postmenopausal Osteoporosis, and to facilitate health care in menopause woman. 3. Effect of soybean isoflavone and calcium on bone mineral density , mineral , BMP₂, TGF-β₁, IGF-1 and IGF-1 mRNA express in ovariectomized rats Objective To study the effect of wheat embryo extract on bone mineral density and mineral in ovariectomized rats.Methods Twenty eight female SD rat were randomly divided into four groups:sham,ovariectomized, ovariectomized added calcium [Ca, 50 mg/kg · d] , ovariectomized added soybean isoflavone [Isof, 100 mg/kg · d], . The experiment proceed ten weeks.Results The bone mineral density decreased, and urine calcium increased significantly in ovariectomized group; The bone BMP₂, TGF-β₁, IGF-1 and IGF-1 mRNA expression in ovariectomized group decreased significantly. The bone mineral density and BMP2, TGF-β₁, IGF-1 and IGF-1 mRNA expression in soybean isoflavone group increased significantly than ovariectomized group; The urine calcium decreased significantly. The recovery of these markers was not significantly in merely supply Calcium group than ovariectomized group.Conclusion The soybean isoflavone may adjustment IGF-1 of growth factor expression, and prevent the lose of calcium of bone and the increase of bone mineral density induced by ovariectomy.4. Effect of soybean isoflavone and calcium on bone mineral density and mineralmatter and kidney TGF β₁ expression in ovariectomized ratsObjective To explore the effect of soybean isoflavone and calcium on bone mineral density and trace elements in ovariectomized rats.Methods Twenty eight female SD rats were randomly divided into four groups:sham, ovariectomized, ovariectomized added calcium (50mg.·kg^-1·d^-1), ovariectomized added soybean isoflavone (100mg.·kg^-1·d^-1).The content of bone mineral density and mineral , and serum, urine and bone calcium, zinc, copper , iron were determined after ten weeks.Results After test , the bone mineral density and bone mineral , and bone calcium, zinc, copper , iron, and serum zinc, iron in ovariectomized group decreased significantly; The serum calcium , copper, ALP and urine calcium, copper, iron increased.The bone mineral density and bone mineral , serum zinc , bone calcium, zinc, copper in ovariectomized added soybean isoflavone group increased significantly than ovariectomized group; The serum calcium, copper, ALP and urine calcium,copper decreased significantly ( P<0.01 or P<0.05 ) . Compare with sham group, theOVX group kidney weight/body weight(%)increased significantly (P<0.01). Comparewith OVX group, the SIFV group kidney weight/body weight(%)decreased significantly(P<0.01) . The glomcrulus TGFβ₁ proteinum expression increased significantly in OVXgroup than sham group and SIFV group(.P<0.01).Conclusion The soybean isoflavone may prevent the lose of bone calcium, zinc ,copper, and restraint the glomcrulus TGFβ₁ proteinum overexpression and prevent theosteoporosis induced by ovariectomy.