淫羊藿苷通过自噬与凋亡途径保护去卵巢大鼠骨质疏松的机制探究  

Mechanism of Icariin-Mediated Autophagy and Apoptosis Pathway on Protecting Ovariectomized Rats Against Osteoporosis

作　　者：何丽君[1] 黄巧文[1] 马黄璜 章靓[1] 张余 王颖峥[2] HE Lijun;HUANG Qiaowen;MA Huanghuang;ZHANG Jing;ZHANG Yu;WANG Yingzheng(People's Hospital Affiliated to Fujian University of Traditional Chinese Medicine,Fuzhou 350004 Fujian,China;Fujian University of Traditional Chinese Medicine,Fuzhou 350122 Fujian,China)

机构地区：[1]福建中医药大学附属人民医院,福建福州350004 [2]福建中医药大学,福建福州350122

出　　处：《中药新药与临床药理》2023年第2期149-155,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：2018年福建省卫计委中青年骨干项目(2018-ZQN-68)。

摘　　要：目的 探讨淫羊藿苷是否通过自噬与凋亡途径保护去卵巢大鼠骨质疏松及其机制。方法 将48只雌性SD大鼠随机分为假手术组、去卵巢组、阳性对照组(尼尔雌醇1 mg·kg^(-1))和淫羊藿苷低、中、高剂量组(40、80、120 mg·kg^(-1)),每组各8只。除假手术组外,各组均进行去卵巢手术。术后灌胃给药12周后,采用双能X线吸收法检测骨密度,采用三点弯曲力学试验测量生物力学指标,采用酶联免疫吸附实验(ELISA)法检测血清骨代谢指标,采用蛋白免疫印迹法(Western Blot)检测自噬相关蛋白P62、自噬效应蛋白(Beclin-1)、LC3-Ⅱ以及凋亡相关蛋白胱天蛋白酶3(Caspase-3)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)和B淋巴细胞瘤基因-2(B-cell lymphoma-2,Bcl-2)表达。结果 淫羊藿苷低、中、高剂量组大鼠骨密度、最大载荷、最大应力以及刚度均高于去卵巢组(P<0.05);淫羊藿苷低、中、高剂量组大鼠血清抗酒石酸酸性磷酸酶(TRAP)、Ⅰ型胶原交联氨基端肽(NTX-Ⅰ)水平均低于去卵巢组(P<0.05),骨钙素(OC)、骨碱性磷酸酶(BALP)水平均高于去卵巢组(P<0.05);淫羊藿苷低、中、高剂量组大鼠骨组织P62表达均低于去卵巢组(P<0.05),Beclin-1、LC3-Ⅱ表达均高于去卵巢组(P<0.05);淫羊藿苷低、中、高剂量组大鼠骨组织Caspase-3、BAX表达均低于去卵巢组(P<0.05),Bcl-2表达均高于去卵巢组(P<0.05)。结论 淫羊藿苷对去卵巢大鼠骨质疏松具有保护作用,其机制可能与增强骨组织自噬水平以及抑制细胞凋亡有关。Objective To explore the mechanism of icariin (ICA)-mediated autophagy and apoptosis pathway on protecting ovariectomized rats against osteoporosis.Methods Forty-eight female SD rats were randomly divided into sham operation group,ovariectomized group,positive control group(1 mg·kg^(-1)of nilestriol),low-,medium-and high-dose of ICA groups(40,80,120 mg·kg^(-1)),with 8 rats in each group.Ovariectomy was performed in each group(except the sham operation group).After surgery,rats were given drugs by intragastric administration for12 weeks.Dual-energy X-ray absorptiometry was used to detect bone mineral density,three-point bending test was adopted to measure the biomechanical indicators.Enzyme-linked immunosorbent assay (ELISA) was applied to detect bone metabolism indicators in serum,and Western Blot was used to detect the expressions of autophagyrelated proteins P62,autophagy effector Beclin-1 and LC3-II and apoptosis-related proteins Caspase-3,Bcl-2associated X protein(Bax)and B-cell lymphoma-2(Bcl-2).Results The bone mineral density,maximum load,maximum stress and stiffness of the rats in the low-,medium-and high-dose of ICA groups were higher than those in the ovariectomized group(P<0.05).The levels of serum tartrate-resistant acid phosphatase(TRAP) and N-telopeptide of type I collagen(NTX-I)in the low-,medium-and high-dose ICA groups were lower than those in the ovariectomized group(P<0.05),while the levels of osteocalcin(OC)and bone alkaline phosphatase(BALP)were higher than those in the ovariectomized group(P<0.05).The expression of P62 in bone tissues in the low-,medium-and high-dose ICA groups was lower than that in the ovariectomized group(P<0.05) while the expressions of Beclin-1 and LC3-II were higher than those in the ovariectomized group(P<0.05).The expressions of Caspase-3 and Bax in bone tissue of the rats in the low-,medium-and high-dose ICA groups were lower(P<0.05)and the expression of Bcl-2 was higher(P<0.05)than those of the ovariectomized group.Conclusion ICA has a protective effect on osteop

关 键 词：淫羊藿苷 去卵巢大鼠 骨质疏松 自噬 凋亡 

分 类 号：R285.5[医药卫生—中药学]