舒芬太尼对去卵巢骨质疏松大鼠骨小梁结构及骨代谢的影响

Effects of sufentanil on bone trabecula structure and bone metabolism in ovariectomized-induced osteoporosis rats

目的探究舒芬太尼对去卵巢骨质疏松大鼠骨小梁结构及骨代谢的影响。方法采用双侧卵巢切除术(OVX)建立绝经后骨质疏松大鼠模型,将50只雌性SD大鼠随机分为5组假手术组、OVX组、OVX+低、中、高剂量舒芬太尼组(1、2、4μg/kg),每组10只。取股骨行uCT扫描测量骨小梁厚度(Tb.Th)、骨小梁分离度(Tb.Sp)、骨体积分数(BV/TV),并观察骨小梁数量及软骨数量;采用HE染色观察股骨组织损伤情况;采用TRAP染色检测破骨细胞数量变化情况;采用实时荧光PCR和蛋白印迹检测骨匀浆液中骨标记蛋白成骨相关转录因子抗体(Osx)、胶原蛋白Ⅰ(Colla1)和远中缺失同源盒基因2(Dlx2)和骨吸收功能相关蛋白(破骨细胞相关受体OSCAR、TRAP)及mRNA的表达。结果与假手术组比较,OVX组骨小梁结构明显破坏,排列无规则,BV/TV、Tb.Th、骨小梁数量、软骨数量及Osx、Colla1、Dlx2 mRNA和蛋白表达均减少/降低(P<0.05),Tb.Sp、破骨细胞数量及OSCAR、TRAP mRNA和蛋白表达均增加/升高(P<0.05);与OVX组比较,OVX+中、高剂量舒芬太尼组骨小梁结构相对完整,排列趋于紧密,BV/TV、Tb.Th、骨小梁数量、软骨数量及Osx、Colla1、Dlx2 mRNA和蛋白表达均增加/升高(P<0.05),Tb.Sp、破骨细胞数量及OSCAR、TRAP mRNA和蛋白表达均减少/降低(P<0.05)。结论舒芬太尼可改善骨小梁微结构、促进骨形成、抑制骨吸收,从而减轻骨质疏松大鼠骨小梁损伤并进行骨重塑。

Objective To explore the effects of sufentanil on bone trabecula structure and bone metabolism in ovariectomized-induced osteoporosis rats.Methods The postmenopausal osteoporosis rat models were constructed by bilateral ovariectomy(OVX).A total of 50 female SD rats were randomly divided into 5 groups:the sham operation group,the OVX group,the OVX+low-dose,medium-dose and high-dose sufentanil groups(1,2,4μg/kg),with 10 rats in each group.The uCT scan for femurs was conducted to measure the trabecular thickness(Tb.Th),trabecular separation(Tb.Sp)and bone volume/total volume(BV/TV).The numbers of bone trabecula and cartilages were observed.The femoral tissue damage was observed by HE staining.The changes in number of osteoclasts were detected by TRAP staining.The expressions of bone marker proteins[osteogenesis-related transcription factor antibody(Osx),collagenⅠ(Colla1),distal-less homeobox(Dlx2)]and bone resorption function related proteins(osteoclast-related receptors OSCAR,TRAP)and mRNA expressions in bone homogenate were detected by real-time fluorescent PCR and Western blot.Results Compared with the sham operation group,the bone trabecular structure was obviously damaged with irregular arrangement in the OVX group,BV/TV,Tb.Th,numbers of bone trabecula and cartilages,and mRNA and protein expression of Osx,Colla1 and Dlx2 were all decreased in the OVX group(P<0.05),while Tb.Sp,number of osteoclasts,and mRNA and protein expression of OSCAR and TRAP were increased in the OVX group(P<0.05).Compared with the OVX group,the bone trabecular structure was relatively intact with dense arrangement in the OVX+medium-dose and high-dose sufentanil groups,BV/TV,Tb.Th,numbers of bone trabecula and cartilages,and mRNA and protein expression of Osx,Colla1 and Dlx2 were all increased in the OVX+medium-dose and high-dose sufentanil groups(P<0.05),while Tb.Sp,numbers of osteoclasts,and mRNA and protein expression of OSCAR and TRAP were decreased in the OVX+medium-dose and high-dose sufentanil groups(P<0.05).Conclusion Sufentanil may reduce trabecular bone damage and conduct bone remodeling in osteoporotic rats by improving microstructure of bone trabecula,promoting bone formation and inhibiting bone resorption.