| Objectibe: To investigate the effects of the decoction of Radix Rehmanniae Preparata(RR), Rhizoma Dioscorea septemlobae(RD), Herba Epimedii(HE) and Radix Astragali(RA) on the bone metabolism in ovariectomized rats and the effects on the proliferation and differentiation of osteoblasts in vitro.Methods: 1.In vivo:35 female, 3-month-old Wistar rats were randomly divided into 4 groups: sham(sham-operation), OVX(ovariectomy), OVX+E(OVX +diethylstilbestrol), OVX+Chinese herbal medicine(OVX+CH), the group of OVX+CH was divided into 4 subgroups: OVX+RR, OVX+RD, OVX+HE and 0VX+RA(n=5 in a group). The rats in OVX+RR, OVX+RD, OVX+HE, OVX+ RA group were given lml/d of the 500mg/ml herbal decoction by gavage, the rats in OVX-E group were given 0.86mg/kg/d diethylstilbestrol dissolved in 1 ml tap water by gavage, and the rats in sham group and OVX were given the tap water by gavage equal volum, once a day, respectively. After 12-week period continuous, the body-weight of all rats was weighted and the urinary samples were collected before they were killed, and then, the blood samples were collected. The samples of urine and blood were taken for determination of serum estrodiol(E2), calcium (Ca), phosphorus(P), bone glaprotein(BGP), alkaline phosphatase(ALP), urinary calcium/creatinine(Ca/Cr), phosphorus/creatinine(P/Cr) and deoxypyridioline/ creatinine(DPD/Cr). The uteri removed and weighed, the bone mineral density(BMD) and the biomechanical parameters of the femur of the rats in all groups were determined, the morphology of the uteri and femur were observed, the parameters of bone morphometry were determined, respectively. 2.In vitro: Neonatal rat calvarial osteoblasts were isolated, the third passage osteoblasts were used. The cells were divided into 2 groups: the serum containing Chinese herbal medicine group and the decoction group. The group of serum containing Chineseherbal medicine was divided into 15 subgroups: 5%, 10%, 15% OVX serum control group; 5%, 10%, 15% serum containing Chinese herbal medicine of RR, RD, HE, RA, respectively. The group of decoction was divided into 13 subgroups: the 1640 culture medium control group,and the different dose of decoction of RR, RD, HE, RA at the concentration of 10, 100, lOOOug/mL, respectively. Cell proliferarion was measured at the 48th hour after culture; the lysate of osteoblasts was used for the measurement of ALP activity and BGP concentration at 96 hour after culture,respectively.Results: I.in vivo: The coefficient of uteri in the OVX group was 0.1843±0.0227(mg/g), significantly lower than that in the sham group(P<0.05); in the group of OVX+RR, OVX+RD, OVX+HE, OVX+RA, the coefficient was 0.277010.0362, 0.5391+0.0820, 0.3183+0.0788, 0.2701±0.0571(mg/g), respectively, it was significantly higher in the group of OVX+RD,OVX+HE than that in the OVX group(P<0.05), and the coefficient in the group of OVX+E was significantly higher than that in the sham group(JP<0.05).The concentration of serum E2 in the OVX group was 13.02±3.59(pg/ml), it was significantly lower than that in the sham group(P<0.05); the concentration in the group of OVX+E was 20.06±4.75(pg/ml), it was significantly higher than that in the OVX group(/><0.05), the concentration of serum E2 was also slightly elevated in the group of OVX+RR, OVX+RD, OVX+HE, OVX+RA, respectively. The concentration of serum ALP, BGP and urinary DPD/Cr, Ca/Cr in the OVX group was 153±20(U/L), 2.73+0.58(ng/ml), 0.64±0.20(umol/L/mmol/L), 0.38±0.12 (mmol/L/mmol/L), it was significantly higher than that in the sham group(P<0.05), respectively; the concentration of serum BGP in the group of OVX+RR, OVX+RD, OVX+HE, OVX+RA was 2.36±0.55, 2.51±0.67, 2.61±0.51, 2.62±0.36 (ng/ml), the concentration of serum ALP in this group was 126±41, 125±44,127±36, 131±34(U/L), it was significantly higher than that in the sham group(P<0.05), respectively; the concentration of urinary Ca/Cr was 0.22±0.05, 0.2010.08, 0.18+0.06, 0.23+0.07(mmol/L/mmol/L), it was lower than in the OVX group,respectively. There was no significantly difference in the concentration of serum Ca, P and urinary P/Cr in every group, respectively. The BMD in the OVX group was 0.032±0.007(g/cm), it was significantly lower than in the sham group(P<0.01); the value was 0.047±0.011, 0.044±0.007, 0.044±0.009, 0.045±0.010(g/cm) in the group of OVX+RR, OVX+RD, OVX+HE, OVX+RA, it was significantly higher than that in the OVX groupf/'O.OS), respectively. The maximum loading, deflection, the maximum strain of the femur in the OVX group was 125.78±15.48(N), 1.87±0.22(mm), 9.34 +1.10(%), it was significantly lower than in the sham group(P<0.05), respectively; the maximum loading and maximum stress was increased in different extent in the group of OVX+RR, OVX+RD, OVX+HE, OVX+RA, respectively. The histological observation of the uteri and the femur tissue showed a atrophic uter in the OVX group, a matked hypertrophic uter in the OVX+E group, and a porotic trabecular bone in the OVX group, respectively. The trabecular bone volume in the OVX group was 25.06±5.28(%), it was significantly lower than in the sham group(PO.Ol); the volume in the group of OVX+RR, OVX+RD, OVX+HE, OVX+RA was 40.36±7.45, 43.48±7.74, 37.71+10.70, 37.04±9.20(%), it was significantly higher than that in the OVX group(P<0.05), respectively. 2. In vitro: At 48 h after culture the OD with MTT in the group of 10%, 15% serum containing Chinese herbal medicine of RR, RD, HE, RA and in the group of lOOug/ml decoction of RR, RD, HE was 0.476±0.077, 0.512+0.064, 0.461±0.076, 0.430±0.044; 0.574±0.052, 0.661±0.054, 0.578±0.051, 0.536±0.092 and 0.403±0.055, 0.432±0.016, 0.392±0.016, it was significantly higher than that in control, respectively^^.05). At 96h after culture...
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