| Objective Examining the circulating concentrations of BALP,OPG, RANKL and the content of BMC to observe the effects of puerarin in IUGR rats. Discussing the effects and the possible mechanism of purarin to IUGR adulthood osteoporosis.Methods1. Model preparation:Eighteen pregnant rats were divided into two groups:①Group with normal protein diet:Six pregnant rats were fed with21%normal protein diet to establish normal birth weight rats;②Group with low protein diet:Twelve pregnant rats were fed with10%low protein diet to establish IUGR rats. Two groups of pregnant rats were fed with21%normal protein diet during lactation period.2. Group:Rats who were from pregnant rats with low protein feed were divided into IUGR model group(model group) and IUGR model intervention group(intervention group); Rats who were from pregnant rats with normal protein feed were divided into normal birth weight group(normal group); During lactation period, all rats were fed with breast milk. After delactacion, all rats were fed with21%normal protein diet.3. Intervention:At the age of lw,intervention group rats were given puerarin by abdominal subcutaneous injection with50mg/kg.d, normal group and model group were given sterilization water with equivalent amount,continuous intervention two weeks.4. Take specimens:At the age1w,3w,8w, weighed weight,killed rats and leaved serum,bone respectively.5. Measurement:serum was collected to assay BALP, OPG and RANKL, which were detected by ELISA at lweek,3week and8week respectively.6.Determine the bone mineral content:At1week,3week and8week,respectively, taking the right tibial on the electric furnace, weighing dry weigh after carbonization, weighing ash weigh after ashing, the ratio of ash weigh and dry weigh was the tibial bone mineral content.7. Statistical analysis:All data were statisticed by the SPSS17.0. Difference between groups were evaluated by variance analysis, rate comparison between groups were evaluated by chi-square test. Significance was set at P<0.05.Results1.Weight:The birth weight of rat in normal group was higher than the model group and intervention group (P＜0.05).At the age of1w, the weight of rat in normal group was higher than the model group and intervention group (P<0.05); At the age of3w, the weight of rat in intervention group was higher than the model group (P<0.05),but lower the normal group (P<0.05); At the age of8w, there was no difference between intervention group and normal groups (P>0.05), higher than the model group (P<0.05)2. Serum makers:(1)Blood calcium and blood phosphorus:There was no difference among three groups(P<0.05).(2)BALP:At the age of lw,the level of BALP in normal group were higher than model group and intervention group (P<0.05); At the age of3w,the level of BALP in model group and intervention group were higher than normal group (P＜0.05); At the age of8w,the level of BALP in intervention group was lower than model group (P＜0.05)(3)OPG,RANKL and OPG/RANKL:At the age of lw,the level of OPG and OPG/RANKL in intervention group and model group were lower than normal group (P<0.05),the level of RANKL were higher than normal group (P＜0.05); there were no differences between intervention group and model group (P>0.05)At the age of3w, compared with model group and normal group, the level of OPG in intervention group was no differences (P>0.05), the level of RANKL was lower (P<0.05),OPG/RANKL was higher than model group (P<0.05);there were no difference between model group and normal group (P>0.05)At the age of8w,the level of RANKL in model group was higher than normal group and intervention group (P<0.05), the level of OPG and OPG/RANKL in intervention group were higher than model group (P<0.05), there was no difference between normal group and intervention group (P>0.05) 3. At the age of1w,the BMC of normal group was higher than model group and intervention group (P＜0.05); At the age of3w, there was no difference among three groups (P>0.05); At the age of8w, the BMC of normal group and intervention group were higher than model group (P＜0.05)Conclusion1. Pregnancy with low-protein diet can lead to intrauterine growth retardation.2. After birth, IUGR rats appears catch-up growth, the puerarin could promote the weight of IUGR rats to increase.3. At the early stage of growth, the level of BALP of IUGR rats increased significantly. Puerarin can reduce the higher bone turnover of IUGR rats.4. Puerarin may be beneficial to the development of skeletal by increasing the expression of OPG,reducing the expression of RANKL and improving the ratio of OPG/RANKL to promote bone formation and increase bone mineral content. |
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