益骨胶囊对绝经后骨质疏松模型大鼠骨组织DBH和ADRB2的影响
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摘要
目的:
观察纯中药复方制剂BBC对PMOP模型大鼠骨组织DBH及ADRB2的表达、分布的变化,探讨PMOP模型大鼠骨组织与交感神经及其递质关系的可能机制,初步研究补肾活血中药复方防治PMOP作用靶点和可能作用机理。
方法:
1.60只10月龄SD雌性大鼠随机分成模型组和假手术组,模型组48只,假手术组12只。模型组大鼠以切除双侧卵巢后饲养12周的方法复制PMOP模型;双能X线吸收测定法测定BMD以确定造模成功。
2.造模12周时将模型组造模成功大鼠再随机分成模型空白组、BBC低剂量组、BBC中剂量组和BBC高剂量组,对BBC低、中、高剂量组大鼠给予相当于成人临床用药量1、3、9倍的BBC药液灌胃12周治疗,假手术组和模型空白组给予相应等量的生理盐水。
3.运用免疫组织化学方法检测各组大鼠股骨远端骨小梁表面细胞的DBH和ADRB2。
4.采用SPSS13.0统计软件,运用秩和检验方法对等级资料进行组间比较。
结果:
1.在造模12周时,模型组和假手术组比较,模型组大鼠多部位骨组织的BMD均较假手术组显著下降(P＜0.05)。
2.造模24周时检测各组大鼠骨组织骨小梁表面细胞ADRB2的表达水平和分布,ADRB2在大鼠股骨远端组织骨髓腔靠近骨小梁表面第一层细胞的细胞膜上表达,模型组和假手术组比较显著上升(P＜0.05);BBC低、中、高剂量组显著低于模型组(P＜0.05);BBC中剂量组和高剂量组间没有显著性差异(P＞0.05)。
3.造模24周时检测各组大鼠骨组织骨小梁表面细胞DBH表达水平,DBH在大鼠股骨远端组织骨髓腔靠近骨小梁表面第一层细胞的细胞上表达,模型空白组和假手术组比较显著上升(P＜0.05);各治疗组间与模型空白组比较无显著性差异(P＞0.05)。
结论:
1.10月龄SD雌性大鼠切除双侧卵巢饲养12周成功复制出PMOP实验动物模型。
2.在PMOP的形成过程中骨组织内交感神经兴奋性提高及表面细胞DBH和ADRB2表达水平增高,可能激活RANKL/RANK/NF-?B信号通路起到了重要的作用。
3.一定剂量依赖性地抑制交感神经兴奋性及降低骨组织表面细胞ADRB2表达水平,可能失活RANKL/RANK/NF-?B信号通路是BBC治疗PMOP的作用机制之一。
Objective:
The research was to observe changes of Benefiting-bone capsule(BBC),a pure Chinese medicine compound preparation,on expression and distribution of dopamine-β-hydroxylase (DBH) andβ2-adrenergic receptor(ADRB2) in cells of bone tissue surface of model rats with postmenopausal osteoporosis(PMOP),explore some possible esoteric mechanism of connection among bone tissue,sympathetic nerve and its neurotransmitters,and preliminary study on targets and the possible mechanisms of Chinese medicine compound preparation treating PMOP.
Methods:
1.60 10-month-old Sprague-Dawley(SD) female rats were divided uniformly into model group and sham group.There are 48 rats in model group and 12 rats in sham group.Rats in the model group were duplicated with the method of breeding for 12 weeks after bilateral ovariectomy.The bone mineral density(BMD) of these rats was checked by Dual energy X-ray absorptiometry(DEXA) to assure that the experimental animal model of osteoporosis was replicated successfully.
2.12 weeks later the rats with model replicated successfully in the model group were divided uniformly into untreated model group,low dose BBC group,middle dose BBC group and high dose BBC group.The rats in the low dose BBC group,middle dose BBC group and high dose BBC group were respectively treated with gastric perfusion of BBC solution that amounted to 1 time,3 times and 9 times dose of BBC used in clinic adult patients,and those in the sham-operated group and untreated model group received gastric perfusion of equal saline, once a day for 12 weeks.
3.Then DBH and ADRB2 in cells on bone trabecula surface of distal femur of rats in every group were checked with immunohistochemistry method.
4.Statistic software SPSS 13.0 was used and rank sum test was adopted for comparison of rank data among groups.
Results:
1.12 weeks after operation the BMD of many bone positions of rats in model group was significantly lower than that of rats in sham-operated group(P＜0.05).
2.24 weeks after operation the expression levels of ADRB2 on the membrane of the first layer of cells in rat distal femur bone marrow cavity near the surface of bone trabecula of untreated model group were significantly higher than that of sham-operated group(P＜0.05); that of low dose BBC group,middle dose BBC group and high dose BBC group were significantly lower than that of untreated model group(P＜0.05);there were not significant difference between middle dose BBC group and high dose BBC group(P＞0.05).
3.24 weeks after operation the expression levels of DBH in the cytoplasm of the first layer of cells in rat distal femur bone marrow cavity near the surface of bone trabecula of untreated model group were significantly higher than that of sham-operated group(P＜0.05);other treated groups were not significantly different from that of untreated model group(P＞0.05).
Conclusions:
1.10-month-old SD female rats had been made successfully into experimental animal model of PMOP by breeding for 12 weeks after bilateral ovariectomy.
2.The increasing excitability of sympathetic nerve and expression of DBH and ADRB2 in cells on bone tissue surface and activing signal pathway of RANKL/RANK/NF-?B in bone tissue may be involved in the abnormality of bone rebuilding and contribute to the morbidity of PMOP.
3.Dose-dependent declining excitability of sympathetic nerve and expression of ADRB2 in cells on bone tissue surface and suppressing signal pathway of RANKL/RANK/NF-?B in bone tissue may be the mechanism of BBC treating PMOP.
观察纯中药复方制剂BBC对PMOP模型大鼠骨组织DBH及ADRB2的表达、分布的变化,探讨PMOP模型大鼠骨组织与交感神经及其递质关系的可能机制,初步研究补肾活血中药复方防治PMOP作用靶点和可能作用机理。
方法:
1.60只10月龄SD雌性大鼠随机分成模型组和假手术组,模型组48只,假手术组12只。模型组大鼠以切除双侧卵巢后饲养12周的方法复制PMOP模型;双能X线吸收测定法测定BMD以确定造模成功。
2.造模12周时将模型组造模成功大鼠再随机分成模型空白组、BBC低剂量组、BBC中剂量组和BBC高剂量组,对BBC低、中、高剂量组大鼠给予相当于成人临床用药量1、3、9倍的BBC药液灌胃12周治疗,假手术组和模型空白组给予相应等量的生理盐水。
3.运用免疫组织化学方法检测各组大鼠股骨远端骨小梁表面细胞的DBH和ADRB2。
4.采用SPSS13.0统计软件,运用秩和检验方法对等级资料进行组间比较。
结果:
1.在造模12周时,模型组和假手术组比较,模型组大鼠多部位骨组织的BMD均较假手术组显著下降(P＜0.05)。
2.造模24周时检测各组大鼠骨组织骨小梁表面细胞ADRB2的表达水平和分布,ADRB2在大鼠股骨远端组织骨髓腔靠近骨小梁表面第一层细胞的细胞膜上表达,模型组和假手术组比较显著上升(P＜0.05);BBC低、中、高剂量组显著低于模型组(P＜0.05);BBC中剂量组和高剂量组间没有显著性差异(P＞0.05)。
3.造模24周时检测各组大鼠骨组织骨小梁表面细胞DBH表达水平,DBH在大鼠股骨远端组织骨髓腔靠近骨小梁表面第一层细胞的细胞上表达,模型空白组和假手术组比较显著上升(P＜0.05);各治疗组间与模型空白组比较无显著性差异(P＞0.05)。
结论:
1.10月龄SD雌性大鼠切除双侧卵巢饲养12周成功复制出PMOP实验动物模型。
2.在PMOP的形成过程中骨组织内交感神经兴奋性提高及表面细胞DBH和ADRB2表达水平增高,可能激活RANKL/RANK/NF-?B信号通路起到了重要的作用。
3.一定剂量依赖性地抑制交感神经兴奋性及降低骨组织表面细胞ADRB2表达水平,可能失活RANKL/RANK/NF-?B信号通路是BBC治疗PMOP的作用机制之一。
Objective:
The research was to observe changes of Benefiting-bone capsule(BBC),a pure Chinese medicine compound preparation,on expression and distribution of dopamine-β-hydroxylase (DBH) andβ2-adrenergic receptor(ADRB2) in cells of bone tissue surface of model rats with postmenopausal osteoporosis(PMOP),explore some possible esoteric mechanism of connection among bone tissue,sympathetic nerve and its neurotransmitters,and preliminary study on targets and the possible mechanisms of Chinese medicine compound preparation treating PMOP.
Methods:
1.60 10-month-old Sprague-Dawley(SD) female rats were divided uniformly into model group and sham group.There are 48 rats in model group and 12 rats in sham group.Rats in the model group were duplicated with the method of breeding for 12 weeks after bilateral ovariectomy.The bone mineral density(BMD) of these rats was checked by Dual energy X-ray absorptiometry(DEXA) to assure that the experimental animal model of osteoporosis was replicated successfully.
2.12 weeks later the rats with model replicated successfully in the model group were divided uniformly into untreated model group,low dose BBC group,middle dose BBC group and high dose BBC group.The rats in the low dose BBC group,middle dose BBC group and high dose BBC group were respectively treated with gastric perfusion of BBC solution that amounted to 1 time,3 times and 9 times dose of BBC used in clinic adult patients,and those in the sham-operated group and untreated model group received gastric perfusion of equal saline, once a day for 12 weeks.
3.Then DBH and ADRB2 in cells on bone trabecula surface of distal femur of rats in every group were checked with immunohistochemistry method.
4.Statistic software SPSS 13.0 was used and rank sum test was adopted for comparison of rank data among groups.
Results:
1.12 weeks after operation the BMD of many bone positions of rats in model group was significantly lower than that of rats in sham-operated group(P＜0.05).
2.24 weeks after operation the expression levels of ADRB2 on the membrane of the first layer of cells in rat distal femur bone marrow cavity near the surface of bone trabecula of untreated model group were significantly higher than that of sham-operated group(P＜0.05); that of low dose BBC group,middle dose BBC group and high dose BBC group were significantly lower than that of untreated model group(P＜0.05);there were not significant difference between middle dose BBC group and high dose BBC group(P＞0.05).
3.24 weeks after operation the expression levels of DBH in the cytoplasm of the first layer of cells in rat distal femur bone marrow cavity near the surface of bone trabecula of untreated model group were significantly higher than that of sham-operated group(P＜0.05);other treated groups were not significantly different from that of untreated model group(P＞0.05).
Conclusions:
1.10-month-old SD female rats had been made successfully into experimental animal model of PMOP by breeding for 12 weeks after bilateral ovariectomy.
2.The increasing excitability of sympathetic nerve and expression of DBH and ADRB2 in cells on bone tissue surface and activing signal pathway of RANKL/RANK/NF-?B in bone tissue may be involved in the abnormality of bone rebuilding and contribute to the morbidity of PMOP.
3.Dose-dependent declining excitability of sympathetic nerve and expression of ADRB2 in cells on bone tissue surface and suppressing signal pathway of RANKL/RANK/NF-?B in bone tissue may be the mechanism of BBC treating PMOP.
引文
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