蛇床子素/壳聚糖衍生物胶束对去卵巢骨质疏松大鼠骨微结构和骨吸收的影响

目的观察蛇床子素/壳聚糖衍生物胶束(Osthole-loaded N-octyl-O-sulfonyl chitosan micelles,NSC-OST)对去卵巢骨质疏松大鼠骨微结构和骨吸收的影响,初步探讨其抗骨质疏松的分子机制。方法将40只雌性SD大鼠按照随机原则分为5组:假手术组(Sham)、模型组(Ovx)、尼尔雌醇组(Nil)、蛇床子素(OST)、蛇床子素/壳聚糖衍生物胶束组(NSC-OST),除了假手术组以外均通过摘除双侧卵巢形成骨质疏松模型,连续给药12周后,通过HE染色和Micro-CT检测分别从二维、三维形态学角度观察药物对骨微结构的影响;通过抗酒石酸酸性磷酸酶染色观察药物对体内破骨细胞生成和分化的影响;通过免疫组化手段检测药物对破骨特异分子NFATc1、c-fos、CTSK表达的影响。结果 HE染色结果显示药物组均能明显改善造模引起的组织骨质疏松样改变,其中Nil组改善骨微结构的效果最明显,其次为NSC-OST组,最后是OST组;通过Micro-CT分析发现NSC-OST可显著提高大鼠股骨的BMD和BV/TV(P0.05),并明显改善Tb.N、Tb.Sp、Tb.Th三项指标(P0.05),效果要优于OST组,但略逊于Nil组; TRAP染色提示NSC-OST可以明显抑制N.Oc/BS和Oc.S/BS两项破骨生成参数(P0.05),效果要优于OST组,但略逊于Nil组;免疫组化实验提示:NSC-OST可以抑制大鼠骨组织中的破骨特异分子NFATc1、c-fos、CTSK的表达,前两项指标NSC-OST与Nil抑制作用相当,抑制CTSK时NSC-OST的作用要弱于Nil,均强于OST。结论 NSC-OST可以显著改善去卵巢大鼠的骨微结构,其抗骨质疏松作用的发挥可能与其抑制破骨细胞的生成和分化相关。

Effects of osthole-loaded N-octyl-O-sulfonyl chitosan micelles on bone microstructures and bone resorption in ovariectomized osteoporotic rats

Objective To observe the effects of osthol-loaded N-octyl-O-sulfonyl chitosan micelles (NSC-OST) on bone microstructures and bone resorption in ovariectomized osteoporotic rats, and to explore its molecular mechanism of anti-osteoporosis. Methods Forty female SD rats were randomly divided into 5 groups: sham group (Sham), model group (Ovx), nilestriol group (Nil), osthol (OST), osthol/chitosan derivative micelle group (NSC-OST). Except for sham operation group, osteoporosis models were established by removing bilateral ovaries. After 12 weeks of continuous administration, osteoporosis models were established. The effects of drugs on bone microstructures was observed with HE staining and micro-CT detection on two-dimensional and three-dimensional morphological perspectives, respectively. The effects of drugs on osteoclast formation and differentiation were observed with tartrate resistant acid phosphatase (TRAP) staining. The effects of drugs on specific molecules NFATc1, c-fos and CTSK expression were detected using immunohistochemistry. Results HE staining results showed that the osteoporosis-like changes induced by modeling in the drug groups improved significantly, and the effect in Nil group was the most obvious, followed by NSC-OST group and OST group. Micro-CT analysis showed that NSC-OST significantly improved BMD and BV/TV of the femur in rats (P< 0.05), and significantly improved Tb. The three indexes Tb.N, Tb.Sp, and Tb.Th (P<0.05) were better than those in OST group, but slightly worse than those in NIL group. TRAP staining showed that NSC-OST significantly inhibited the osteoclast formation parameters N.Oc/BS and Oc.S/BS (P<0.05), and the effect was better than that in OST group, but slightly worse than that in NIL group. Immunohistochemical results showed that NSC-OST inhibited the expression of osteoclast-specific molecules NFATc1, c-fos, and CTSK. The inhibitive effect on the first two indicators was similar in NSC-OST group and in Nil group. When CTSK was inhibited, the efficacy in NSC-OST group was less than that in Nil group but more than that in OST group. Conclusion NSC-OST significantly improves the bone microstructures in ovariectomized rats, and its anti-osteoporosis effect may be related to the inhibition of osteoclast formation and differentiation.