淫羊藿黄酮对被动吸烟大鼠骨量与骨代谢生化指标的影响及其相关性研究

目的 探讨淫羊藿黄酮干预对被动吸烟大鼠骨量与骨代谢生化指标的影响及其相关性。方法选用2月龄Sprague—Dawley（SD）大鼠60只，随机分为6组，每组10只，各组雌雄各半，雌雄分笼饲养。A组：空白对照组，常规方法饲养，不给予被动吸烟；B组：被动吸烟组，不给予灌服药物；C组：被动吸烟＋钙组，灌服高效钙（75mg·ks^-1·d^-1）＋维生素D321IU·kg^-1-d^-1；D组：被动吸烟＋低剂量灌服淫羊藿黄酮（75mg·kg^-1·d^-1）]；E组：被动吸烟＋中剂量灌服淫羊藿黄酮（150mg·kg^-1·d^-1）]；F组：被动吸烟＋高剂量灌服淫羊藿黄酮（300mg·kg^-1·d^-1）]。按“密室熏烟法”给予实验的（B、C、D、E、F组）大鼠被动吸烟4个月。实验动物干预8周和4个月时测定血Ca、血P、血ALP、尿Ca、尿P、血清骨钙素、抗酒石酸酸性磷酸酶、尿脱氧吡啶啉、股骨和腰椎骨密度，并分析骨密度与骨代谢生化指标之间的相关性。结果（1）股骨和腰椎骨密度、BGP在第8周和在4个月末时血ALP指标A、C、D、E、F组较B组为高，其差异有统计学意义（P〈0．05）；D、E、F组随淫羊藿黄酮剂量的增加而增加，但其相互之间差异元统计学意义（P〉0．05）；其它各组之间相互比较其差异无统计学意义（P〉0．05）。TRACP的活性、Dpd浓度、在第8周和在4个月末时尿Ca、尿Hop指标A、C、D、E、F组较B组为低，其差异有统计学意义（P〈0．05）；D、E、F组随淫羊藿黄酮剂量的增加而降低，但其相互之间差异无统计学意义（P〉0．05）；其它各组之间相互比较其差异无统计学意义（P〉0．05）。（2）在第8周及在4个月末时尿Ca、尿Hop和TRACP及Dpd／Cr指标A、C、D、E、F组较B组为低，其差异有统计学意义（P〈0．05）；D、E、F组随淫羊藿黄酮剂量的增加而降低，但其相互之间差异无统计学意义（P〉0．05）；其它各组之间相互比较其差异无统计学意义（P〉0．05）。股骨和腰椎骨密度与BGP、ALP呈明显正相关（r=0．784及0．816，P〈0．05；r=0．743及0．807，P〈0．05）。股骨和腰椎骨密度与TRACP的活性、Dpd浓度、尿Ca、尿Hop呈明显负相关（r分别=-0．617、-0．608、-0．587、-0．611，P〈0．05；r分别=-0．614、-0．621、-0．583、-0．617，P〈0．05）。结论淫羊藿黄酮干预可明显改善被动吸烟大鼠的骨密度和骨生化指标，且骨密度与骨生化指标之间存在明显的相关性。

Effect of EPF on the bone mineral density and biochemical markers of bone metabolism in the passive smoking rats and the correlations between BMD and biochemical markers

Objective To explore the pharmacological effects of epimedium pubeseen flavonoid （EPF） on the hone mineral density（ BMD）, the biochemical markers of hone metabolism in the passive smoking rats, and the correlations between BMD and the biochemical markers. Methods Sixty Sprague-Dawley （SD） rats （ aged 2-monthold） were randomly divided into six groups（n = 10） ,including sham control group（group A） ,model group（group B） ,eontrol group（group C） ,treated 1 group（group D） ,treated 2 group（group E） and treated 3 group（group F）. There were five male rats and five female rats in each group, and the male and female rats had been fed in different cage. All the rats were given the same feedstuff. Meanwhile, group C were given calcium 75 mg · kg^-1· d^-1 com bined with WitD3 21 IU · kg^-1· d^-1 by gastrogavage for4 months; group D EPF 75 mg · kg^-1· d^-1group E EPF 150 mg · kg^-1· d^-1; group F EPF 300 mg · kg^-1· d^-1. Except the sham control group（group A）, the other groups had been exposed daily to passive cigarette smoking for four months basing on the adytum fuming method. The experimental rats＇ urine in 24 hours had been collected through the metabolize cage means and the vein blood had been collected through to cut off the rats＇ trail at the end of the eighth weeks and the fourth months. These had been measured i. e. （1）The serum Ca,serum serum ALP,urine Ca,urine P,and urine Hop at the end of the eighth week and the fourth month; （2）The Serum BGP,Serum TRACP,Urine Dpd/Cr; （3） the BMD of the femur and lum bar vertebra body. The correlations had been analyzed between the BMD and the biochemical markers of the bone for mation and resorption. Results （ 1 ） The BMD of the femur and lumbar vertebra body, the serum BGP（ng/ml）, the serum ALP at the end of the eighth weeks and at the end of the fourth months had increased markedly in group A, group C, group D,group E and group F compared with group B, and the difference between group A, group C, group D,group E,group F and group B had statistical significance（P 〈0.05）. There had dose-dependently increase in group D,group E and group F, and the difference among them had no statistical signifieance（P 〉0. 05）. The differenee had no statistical significance（P 〉0. 05） among the other groups. The serum TRACP（U/L） and urine Dpd/Cr （nmol/mmol） had decreased markedly in group A,group C, group D,group E and group F compared with group B, and the difference between group A, group C, group D, group E, group F and group B had statistical significance（ P 〈 0. 05）. There had dose-dependently decreased in group D,group E and group F, and the difference among them had no statistical significance （ P 〉 0. 05 ）. The difference had no statistical significance （ P 〉 0. 05 ） among the other groups; （2） The urine Ca and urine Hop at the end of the fourth months and at the end of the eighth weeks, the serum TRACP and urine Dpd/Cr had decreased markedly in group A, group C, group D, group E and group F compared with group B, and the difference between group A,group C,group D,group E,greup F and group B had statistical significance（ P 〈 0. 05 ）. There had dose-dependently decreased in group D, group E and group F, and the difference among them had no statistical significance （P 〉 0. 05 ）. The difference had no statistical significance （P 〉 0.05 ） among the other groups. The serum ALP had increased markedly in group A, group C, group D, group E and group F compared with group B, and the difference between group A,group C,group D,group E,group F and group B had statistical significance（P 〈0. 05） at the end of the eighth weeks and at the end of the fourth months. There had dose-dependently decreased in group D, group E and group F, and the difference among them had no statistical significance （P 〉 0. 05 ）. The difference had no statistical significance （P 〉 0. 05 ） among the other groups. The difference of the other biochemical indices had no statistical significance （ P 〉 0. 05 ） among the other groups. A positive correlation was found between the BMD of the femur and lumbar vertebra body and serum BGP, serum ALP （r=0. 784,0. 816, P 〈 0. 05 ;r = 0. 743,0. 807, P 〈 0. 05 ）. There were also a negative correlation between the BMD of the femur and lumbar vertebra body and serum TRACP, Urine Dpd/Cr, urine Ca, urine Hop（ r = - 0. 617, - 0. 608, - 0. 587, - 0.611,P〈0.05;r = -0.614, -0.621, -0.583, -0.617,P 〈0.05）. Conclusion The EPF could prevent and treat effectively the decreasing of BMD and the worsening of biochemical markers of bone metabolism resulted from passive smoking in the experimental rats. There are significant correlations between the BMD and the biochemical markers of the bone formation and resorption.