瑞舒伐他汀通过AMPK/Sirt1/NF-κB通路改善骨质疏松雌性大鼠骨微结构的研究  

Effects of rosuvastatin on AMPK/Sirt1/NF-κB pathway and bone microstructure in rats with osteoporosis

作　　者：牛建锋[1] 朱海慧 杨庆宇[1] 刘建华 田明波[2] NIU Jian-feng;ZHU Hai-hui;YANG Qing-yu;LIU Jian-hua;TIAN Ming-bo(Department of Pharmacy,People’s Hospital of Zhengzhou,Zhengzhou 450003,China;Department of orthopedics,People’s Hospital of Zhengzhou,Zhengzhou 450003,China)

机构地区：[1]郑州人民医院药学部,河南郑州450003 [2]郑州人民医院骨科,河南郑州450003

出　　处：《现代药物与临床》2022年第1期25-32,共8页Drugs & Clinic

基　　金：河南省医学科技攻关项目(202102310507)。

摘　　要：目的探讨瑞舒伐他汀对骨质疏松大鼠腺苷酸活化蛋白激酶(AMPK)/沉默信息调节因子1(Sirt1)/核转录因子-κB(NF-κB)通路和骨微结构的影响。方法将雌性SD大鼠随机分为假手术组、模型组、雌二醇(0.05 mg/kg)组、瑞舒伐他汀(1.0、2.0、4.0 mg/kg)组,每组12只。采用双侧卵巢切除术复制大鼠骨质疏松症模型。术后第9周雌二醇组和瑞舒伐他汀各剂量组ip相应药物,1次/d,连续给药12周。假手术组和模型组ip等体积生理盐水。ELISA法检测血清骨代谢标志物成骨特异性转录因子(CBF-α1)、Ⅰ型胶原交联羧基端肽(CTXI)、Ⅰ型前胶原氨基端原肽(PINP)、骨钙素(OC)水平和血清雌二醇(E;)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)水平;Micro-CT扫描大鼠右侧股骨观察大鼠股骨远端骨微结构变化,分析骨小梁骨密度(BMD)、骨体积分数(BV/TV)、骨小梁数量(Tb.N)、骨小梁间隔(Tb.Sp)、骨小梁厚度(Tb.Th);苏木精–伊红(HE)染色观察骨组织形态学变化;Western blotting法检测骨组织Sirt1、AMPK、p-AMPK、NF-κB p65、NF-κBp65(acetyl K310)蛋白表达。结果与假手术组相比,模型组大鼠血清E;、骨代谢相关标志物CBF-α1、CTX-Ⅰ、PINP、OC水平、BMD明显降低,炎症因子IL-6、TNF-α、IL-1β水平明显升高,骨小梁微结构参数BV/TV、Tb.N、Tb.Th明显降低,Tb.Sp明显升高(P<0.05),骨小梁出现断裂,排列疏松,骨组织pAMPK/AMPK、Sirt1蛋白表达明显降低,NF-κB p65(acetyl K310)/NF-κB p65蛋白表达明显升高(P<0.05);与模型组相比,瑞舒伐他汀2.0、4.0 mg/kg组大鼠血清E;、骨代谢相关标志物CBF-α1、CTX-Ⅰ、PINP、OC水平、BMD明显升高(P<0.05),炎症因子IL-6、TNF-α、IL-1β水平下降,骨小梁微结构参数BV/TV、Tb.N、Tb.Th增加,Tb.Sp明显降低(P<0.05),新生骨小梁,形态相对完整;骨组织p-AMPK/AMPK、Sirt1蛋白表达明显升高,NF-κB p65(acetyl K310)/NF-κB p65蛋白表达明显降低(P<0.05)。�Objective To investigate the effects of rosuvastatin on AMP-activated protein kinase(AMPK)/silent information regulator 1(Sirt1)/nuclear factor-κB(NF-κB) pathway and bone microstructure in osteoporosis rats. Methods Female SD rats were randomly divided into sham operation group, model group, estradiol(0.05 mg/kg) group and rosuvastatin(1.0, 2.0, and 4.0 mg/kg) groups, with 12 rats in each group. Osteoporosis(OP) model was established by bilateral ovariectomy. At 9 th week after operation, the estradiol group and rosuvastatin groups were ip administered with corresponding drugs, once daily for 12 weeks. The sham operation group and model group were ip administered with equal volume of normal saline. After 12 weeks of administration, the samples were collected and related indexes were detected. The levels of serum core-binding factor α1(CBF-α1), cross linked C-telopeptide of type I collagen(CTXI), procollagen I N-terminal propeptide(PINP), osteocalcin(OC), estradiol(E;), interleukin-6(IL-6), tumor necrosis factor(TNF-α), and interleukin-1β(IL-1β) were detected by ELISA method. The right femur of rats was scanned by Micro-CT, the changes of bone microstructure of distal femur were observed, and trabecular bone mineral density(BMD), bone volume fraction(BV/TV), trabecular number(Tb. N), trabecular septum(Tb.Sp), and trabecular thickness(Tb.Th) were analyzed. Hematoxylin eosin(HE) staining was used to observe the morphological changes of bone tissue. The protein expression of Sirt1, AMPK, p-AMPK, NF-κB p65, and NF-κB p65(acetyl K310) was detected by Western blotting method. Results Compared with those in the sham operation group, the serum E;, CBF-α1, CTX-Ⅰ, PINP, OC, and BMD were significantly lower in the model group, the levels of inflammatory factors IL-6, TNF-α, and IL-1β were significantly higher, bone trabecular microstructure parameters BV/TV, Tb.N, and Tb.Th were significantly lower,Tb.Sp was significantly higher(P < 0.05), while the trabeculae were broken and arranged loosely, the protein expre

关 键 词：瑞舒伐他汀 骨质疏松症 腺苷酸活化蛋白激酶/沉默信息调节因子1/核转录因子-κB 骨微结构 保护作用 

分 类 号：R965[医药卫生—药理学]