目的 探討瑞舒伐他汀對骨質(zhì)疏松大鼠腺苷酸活化蛋白激酶（AMPK）/沉默信息調(diào)節(jié)因子1（Sirt1）/核轉(zhuǎn)錄因子-κB（NF-κB）通路和骨微結(jié)構(gòu)的影響。方法 將雌性SD大鼠隨機分為假手術(shù)組、模型組、雌二醇（0.05 mg/kg）組、瑞舒伐他?。?.0、2.0、4.0 mg/kg）組，每組12只。采用雙側(cè)卵巢切除術(shù)復(fù)制大鼠骨質(zhì)疏松癥模型。術(shù)后第9周雌二醇組和瑞舒伐他汀各劑量組ip相應(yīng)藥物，1次/d，連續(xù)給藥12周。假手術(shù)組和模型組ip等體積生理鹽水。ELISA法檢測血清骨代謝標志物成骨特異性轉(zhuǎn)錄因子（CBF-α1）、Ⅰ型膠原交聯(lián)羧基端肽（CTXI）、Ⅰ型前膠原氨基端原肽（PINP）、骨鈣素（OC）水平和血清雌二醇（E₂）、白細胞介素6（IL-6）、腫瘤壞死因子α（TNF-α）、白細胞介素1β（IL-1β）水平；Micro-CT掃描大鼠右側(cè)股骨觀察大鼠股骨遠端骨微結(jié)構(gòu)變化，分析骨小梁骨密度（BMD）、骨體積分數(shù)（BV/TV）、骨小梁數(shù)量（Tb.N）、骨小梁間隔（Tb.Sp）、骨小梁厚度（Tb.Th）；蘇木精–伊紅（HE）染色觀察骨組織形態(tài)學(xué)變化；Western blotting法檢測骨組織Sirt1、AMPK、p-AMPK、NF-κB p65、NF-κ Bp65（acetyl K310）蛋白表達。結(jié)果 與假手術(shù)組相比，模型組大鼠血清E₂、骨代謝相關(guān)標志物CBF-α1、CTX-Ⅰ、PINP、OC水平、BMD明顯降低，炎癥因子IL-6、TNF-α、IL-1β水平明顯升高，骨小梁微結(jié)構(gòu)參數(shù)BV/TV、Tb.N、Tb.Th明顯降低，Tb.Sp明顯升高（P＜0.05），骨小梁出現(xiàn)斷裂，排列疏松，骨組織p-AMPK/AMPK、Sirt1蛋白表達明顯降低，NF-κB p65（acetyl K310）/NF-κB p65蛋白表達明顯升高（P＜0.05）；與模型組相比，瑞舒伐他汀2.0、4.0 mg/kg組大鼠血清E₂、骨代謝相關(guān)標志物CBF-α1、CTX-Ⅰ、PINP、OC水平、BMD明顯升高（P＜0.05），炎癥因子IL-6、TNF-α、IL-1β水平下降，骨小梁微結(jié)構(gòu)參數(shù)BV/TV、Tb.N、Tb.Th增加，Tb.Sp明顯降低（P＜0.05），新生骨小梁，形態(tài)相對完整；骨組織p-AMPK/AMPK、Sirt1蛋白表達明顯升高，NF-κB p65（acetyl K310）/NF-κB p65蛋白表達明顯降低（P＜0.05）。結(jié)論 瑞舒伐他汀可改善骨質(zhì)疏松大鼠的骨微結(jié)構(gòu)，對骨質(zhì)疏松具有保護作用，其作用機制可能與激活A(yù)MPK和Sirt1蛋白表達，降低NF-κB p65乙?；接嘘P(guān)。

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 9th 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 (E₂), 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 E₂, 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 expression of p-AMPK/AMPK and Sirt1 in bone tissue was significantly lower, and the protein expression of NF-κB p65 (acetyl K310)/NF-κB p65 was significantly higher (P < 0.05). Compared with those in the model group, the serum E₂, CBF-α1, CTX-Ⅰ, PINP, OC, and BMD were significantly higher in the rosuvastatin 2.0 and 4.0 mg/kg groups, the levels of inflammatory factors IL-6, TNF-α, and IL-1β were significantly lower, bone trabecular microstructure parameters BV/TV, Tb.N, and Tb.Th were significantly higher, while Tb.Sp was significantly lower (P < 0.05), and new bone trabeculae could be seen with relatively complete shape. The protein expression of p-AMPK/AMPK and Sirt1 in bone tissue was significantly higher, but the protein expression of NF-κB p65 (acetyl K310)/NF-κB p65 was significantly lower (P < 0.05).Conclusion Rosuvastatin can improve the bone microstructure and protect osteoporosis in rats, which may be related to the activation of AMPK and Sirt1 protein expression and the decrease of NF-κB p65 acetylation level.

R965

河南省醫(yī)學(xué)科技攻關(guān)項目（202102310507）