秦皮对去卵巢所致大鼠骨质疏松症的治疗作用及机理探讨
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摘要
目的:初步探讨秦皮治疗骨质疏松症的作用机理。方法:健康雌性SD大鼠40只按随机数字表法分为空白对照组、假手术组各10只、造模组20只,造模组大鼠行双侧卵巢切除术制作骨质疏松症模型,造模结束后再将造模组大鼠按随机数字表法分为模型组和秦皮组各10只,开始给予相应药物。给药3个月后取左侧胫骨检测骨组织形态计量学指标,取右侧胫骨检测大鼠胫骨骨髓中OPG和RANKL的蛋白表达。结果:模型组大鼠胫骨TBV%、胫骨骨髓中OPG蛋白表达显著低于假手术组,而RANKL蛋白表达及TRS%、TFS%、OSW和MAR均明显高于假手术组。与模型组比较,秦皮组大鼠胫骨TBV%、OPG蛋白表达明显增高,但仍显著低于假手术组;而RANKL蛋白表达、TRS%、TFS%、OSW和MAR均明显降低。结论:秦皮对去卵巢所致的大鼠骨质疏松症具有一定的治疗作用,其作用途径是对OPG/RANKL通路进行调节,促进OPG蛋白表达而抑制RANKL蛋白表达,从而抑制过度增强的破骨细胞活性,防止骨量的快速丢失。
Objective: Explore the effect and mechanism of Cortex Fraxini in treatment osteoporosis. Methods: Forty SPF female SD rats were randomly divided into three groups: blank group( n = 10),sham operation group( n = 10) and modeling group( n = 20). Rats in the modeling group were treated with bilateral ovariectomized( OVX) to make osteoporosis models. Ten days after surgery,the rats in the modeling group were divided into two groups: Model group and Cortex Fraxini Group,10 rats in each group. Then feed the drug and water to rats of each group accordingly. After three months of drugs feeding,we measured the histomorphometry indexes of rats tibia,and the protein expression of OPG and RANKL. Results: Compared with sham operation group,the TBV% of the tibia in Model group was significantly decreased,well the TRS%,TFS%,OSW and MAR were increased obviously. Compared with Model group,the TBV% of the tibia was significantly increased in Cortex Fraxini group,the TRS%,TFS%,OSW and MAR were decreased obviously. Compared with sham operation group,the protein expression of OPG decreased significantly but the protein expression of RANKL increased significantly in model group. The expression of OPG protein in Cortex Fraxini group were significantly higher than those in the model group,while the expression of RANKL was significantly lower than that of model group,but there was still a significant difference compared with the sham operation group. Conclusion: The prescription of Cortex Fraxini has therapeutic effects on osteoporosis rats. The common pathway is that Cortex Fraxini can regulate the expression of OPG and RANKL,the expression of OPG up-regulated and RANKL down-regulated. Thus,the osteoclast activity was inhibited and the rapid loss of bone mass was prevented.
Objective: Explore the effect and mechanism of Cortex Fraxini in treatment osteoporosis. Methods: Forty SPF female SD rats were randomly divided into three groups: blank group( n = 10),sham operation group( n = 10) and modeling group( n = 20). Rats in the modeling group were treated with bilateral ovariectomized( OVX) to make osteoporosis models. Ten days after surgery,the rats in the modeling group were divided into two groups: Model group and Cortex Fraxini Group,10 rats in each group. Then feed the drug and water to rats of each group accordingly. After three months of drugs feeding,we measured the histomorphometry indexes of rats tibia,and the protein expression of OPG and RANKL. Results: Compared with sham operation group,the TBV% of the tibia in Model group was significantly decreased,well the TRS%,TFS%,OSW and MAR were increased obviously. Compared with Model group,the TBV% of the tibia was significantly increased in Cortex Fraxini group,the TRS%,TFS%,OSW and MAR were decreased obviously. Compared with sham operation group,the protein expression of OPG decreased significantly but the protein expression of RANKL increased significantly in model group. The expression of OPG protein in Cortex Fraxini group were significantly higher than those in the model group,while the expression of RANKL was significantly lower than that of model group,but there was still a significant difference compared with the sham operation group. Conclusion: The prescription of Cortex Fraxini has therapeutic effects on osteoporosis rats. The common pathway is that Cortex Fraxini can regulate the expression of OPG and RANKL,the expression of OPG up-regulated and RANKL down-regulated. Thus,the osteoclast activity was inhibited and the rapid loss of bone mass was prevented.
引文
[1]鞠大宏,张丽坤,赵红艳,等.滋阴补肾法对卵巢切除所致骨质疏松大鼠骨髓基质细胞IL-1和成骨细胞IL-6表达的影响[J].中国中医基础医学杂志,2005,11(7):20-23.
[2]鞠大宏,于福禄,张丽坤,等.滋阴补肾法对卵巢切除所致骨质疏松大鼠成骨细胞COX-2蛋白和Mrna表达的影响[J].中国中医基础医学杂志,2006,14(12):44-46.
[3]张颖,Gary Guishan Xiao,荣培晶,等.杜仲、千年健对去卵巢大鼠骨质疏松症的治疗作用及其机理探讨[J].中国中医基础医学杂志,2011,17(9):960-962.
[4]方莲花,吕扬,杜冠华.秦皮的药理作用进展[J].中国中药杂志,2008,33(23):2732-2736.
[5]赫军,马秉智,赵铁,等.藤梨根的化学成分研究[J].中国药学杂志,2014,49(3):184-186.
[6]刘世清,贺翎,彭昊,等.秦皮对兔实验性骨关节炎的基质金属蛋白酶-1和一氧化氮及前列腺素E2的作用[J].中国临床康复,2005,9(6):150.
[7] JIMENEZ-OROZCO FA,ROSALES AA,VEGA-LOPEZ A,etal. Differential effects of esculetin and daphnetin on in vitro cellproliferation and in vivo estrogenicity[J]. Eur J Pharmacol,2011,668(1-2):35-41.
[8] YAMADA H, WATANABE K, SAITO T, et al. Esculetin(dihydroxycoumarin)inhibits the production of matrixmetalloproteinases in cartilage explants,and oral administrationof its prodrug, CPA-926, suppresses cartilage destruction inrabbit experimental osteoarthritis[J]. J Rheumatol,1999,26(3):654-662.
[9] KUO P L,HUANGY T,CHANG C H,et a.l Fraxetin inhibitsthe induction of anti-Fas Ig M,tumor necrosis factor-alpha andinter-leukin-lbeta-mediated apoptosis by Fas pathway inhibitionin hu-man osteoblastic cell line MG-63[J]. IntImmunopharmaco,2006,6(7):1167.
[10] KUO P L, HUANG Y T, CHANG C H, et a. l Bonemorphogenetic protein-2 and-4(BMP-2 and-4)mediatesfraxetin-induced mat-uration and differentiation in humanosteoblast-like cell lines[J]. BiolPharm Bull,2006,29(1):119.
[11]刘梅洁,李鸿泓,王少君,等.骨质疏松症脾肾两虚型病证结合动物模型的建立[J].中国中医基础医学杂志,2012,18(8):836-838.
[12]董佳梓,鞠大宏,贾朝娟,等.桑寄生、枸杞子、桑椹对去卵巢大鼠骨质疏松症的治疗作用及其机理探讨[J].2010,16(6):483-486.
[13]车路阳,郭志兰,郭清华,等.优化培养条件和裂解方法提高hRANKL在大肠杆菌内的表达[J].生物工程学报,2017,33(5):849-862.
[2]鞠大宏,于福禄,张丽坤,等.滋阴补肾法对卵巢切除所致骨质疏松大鼠成骨细胞COX-2蛋白和Mrna表达的影响[J].中国中医基础医学杂志,2006,14(12):44-46.
[3]张颖,Gary Guishan Xiao,荣培晶,等.杜仲、千年健对去卵巢大鼠骨质疏松症的治疗作用及其机理探讨[J].中国中医基础医学杂志,2011,17(9):960-962.
[4]方莲花,吕扬,杜冠华.秦皮的药理作用进展[J].中国中药杂志,2008,33(23):2732-2736.
[5]赫军,马秉智,赵铁,等.藤梨根的化学成分研究[J].中国药学杂志,2014,49(3):184-186.
[6]刘世清,贺翎,彭昊,等.秦皮对兔实验性骨关节炎的基质金属蛋白酶-1和一氧化氮及前列腺素E2的作用[J].中国临床康复,2005,9(6):150.
[7] JIMENEZ-OROZCO FA,ROSALES AA,VEGA-LOPEZ A,etal. Differential effects of esculetin and daphnetin on in vitro cellproliferation and in vivo estrogenicity[J]. Eur J Pharmacol,2011,668(1-2):35-41.
[8] YAMADA H, WATANABE K, SAITO T, et al. Esculetin(dihydroxycoumarin)inhibits the production of matrixmetalloproteinases in cartilage explants,and oral administrationof its prodrug, CPA-926, suppresses cartilage destruction inrabbit experimental osteoarthritis[J]. J Rheumatol,1999,26(3):654-662.
[9] KUO P L,HUANGY T,CHANG C H,et a.l Fraxetin inhibitsthe induction of anti-Fas Ig M,tumor necrosis factor-alpha andinter-leukin-lbeta-mediated apoptosis by Fas pathway inhibitionin hu-man osteoblastic cell line MG-63[J]. IntImmunopharmaco,2006,6(7):1167.
[10] KUO P L, HUANG Y T, CHANG C H, et a. l Bonemorphogenetic protein-2 and-4(BMP-2 and-4)mediatesfraxetin-induced mat-uration and differentiation in humanosteoblast-like cell lines[J]. BiolPharm Bull,2006,29(1):119.
[11]刘梅洁,李鸿泓,王少君,等.骨质疏松症脾肾两虚型病证结合动物模型的建立[J].中国中医基础医学杂志,2012,18(8):836-838.
[12]董佳梓,鞠大宏,贾朝娟,等.桑寄生、枸杞子、桑椹对去卵巢大鼠骨质疏松症的治疗作用及其机理探讨[J].2010,16(6):483-486.
[13]车路阳,郭志兰,郭清华,等.优化培养条件和裂解方法提高hRANKL在大肠杆菌内的表达[J].生物工程学报,2017,33(5):849-862.