仙灵骨葆预防卵巢切除大鼠的腰椎间盘退变

Xianling Gubao prevents lumbar intervertebral disc degeneration in ovariectomized rats

背景:仙灵骨葆主要成分为黔岭淫羊藿、补骨脂、断续等组成,临床上常被用于治疗骨质疏松症。目的:验证仙灵骨葆对去卵巢大鼠椎间盘退变预防作用并探讨其作用机制。方法:3月龄雌性SD大鼠被随机均分为3组。双侧卵巢切除+仙灵骨葆组切除双侧卵巢后,250mg/(kg?d)仙灵骨葆灌胃,1次/d,连续用药12周。假手术组暴露卵巢但不切除;双侧卵巢切除+盐水组切除双侧卵巢后给予等量生理盐水。以上3组均于术后3个月处死,并于处死前10d和4d分别给予四环素和钙黄绿素双荧光标记。取L2椎体进行骨形态计量学分析,L4～L5腰椎及椎间盘进行VG特殊染色及采用免疫组织化学检测椎间盘基质金属蛋白酶13的表达,观察椎间盘的病理学改变并根据组织学评分系统对腰椎间盘的退变程度(LVD)进行评分。结果与结论:①腰椎骨量:双侧卵巢切除+盐水组显著低于假手术组及双侧卵巢切除+仙灵骨葆组(P<0.05)。②腰椎间盘评分:双侧卵巢切除+盐水组组显著高于假手术组及双侧卵巢切除+仙灵骨葆组(P<0.05)。③免疫组织化学:双侧卵巢切除+盐水组与双侧卵巢切除+仙灵骨葆组、假手术组比较大鼠椎间盘中基质金属蛋白酶13表达显著升高(P<0.05)。结果表明,仙灵骨葆可预防去卵巢大鼠腰椎间盘的退变,其作用机制可能包括维持椎体骨量及微观结构、抑制终板退变从而改善椎间盘的生物力学环境和营养供应,同时降低椎间盘中基质金属蛋白酶13的表达,抑制基质的降解。

BACKGROUND：Xianling Gubao（XLGB） mainly contains icariin,buguzhi,duanxu and so on,and it has been widely used to treat osteoprosis in clinic.OBJECTIVE：To investigate the effects of XLGB on lumbar intervertebral disc degeneration（LVD） in the ovariectomized（OVX） rats.METHODS：Thirty 3-month-old female Sprague-Dawley rats were randomly divided into 3 groups.Bilateral OVX ＋ XLGB group was treated with an intragastric perfusion of 250 mg/（kg？d） XLGB,once a day,for 12 successive weeks.Ovary was exposed but not ovariectomized in the sham-surgery group.Bilateral OVX ＋ saline group was treated with an equal volume of saline.All the rats were sacrificed after 3 months postsurgery.Double labeling was performed with subcutaneous injections of tetracylcin 10 days and of calcein 4 days before sacrifice.Bone histomorphometric analysis was applied to the L2 lumbar spines in all rats.L4-5 lumbar spines and intervertebral disc sections were stained by collagen VG staining and immunostained for MMP-13 expression.The pathological changes of intervertebral discs were observed and degree of lumbar intervertebral disc degeneration（LVD） was scored by a histological scoring system.RESULTS：① Bone histomorphometric analysis：Spine bone mass in the OVX＋ saline group was significantly lower than that in sham-surgery and OVX＋XLGB groups（P〈0.05）.② Histological scoring：Rats in OVX＋ saline group had significantly higher scores than those in sham-surgery and OVX＋XLGB group（P〈0.05）.③ Immunostaining：MMP-13 expression in the OVX＋ saline group was significantly higher than that in sham-surgery and OVX＋XLGB groups（P〈0.05）.XLGB could prevent lumbar intervertebral disc degeneration in the OVX rats,probably from maintaining bone mass and microarchitecture of vertebral body and inhibiting endplate calcifaction thereby improving the biomechanical environment and nutrition supply of intervertebral disc,as well as counteracting extracellular matrix degradation by down-regulation of MMP-13 expression.