巴戟天对骨质疏松破骨细胞表面Ⅰ型跨膜受体蛋白的影响

Effects of Morinda on receptor activator of nuclear factor-kappa B expression in osteoclasts in osteoporosis

背景:研究显示巴戟天能够直接刺激体外成骨细胞增加,促进成骨细胞分泌碱性磷酸酶和骨钙素,促进成骨细胞表达转化生长因子β1mR NA。但是,对于巴戟天对骨质疏松大鼠破骨细胞RANK的影响机制尚缺乏报道。目的:分析巴戟天对骨质疏松大鼠破骨细胞表面的Ⅰ型跨膜受体蛋白的影响机制。方法:采用随机对照方法将30只SD大鼠分为巴戟天组和17β-雌二醇组,每组15只。采用腹腔注入盐酸氯胺酮麻醉,切除双侧卵巢,建立大鼠骨质疏松动物模型,造模后大鼠正常饮食。17β-雌二醇组灌胃给予5 m L浓度10-6 mmol/L的17β-雌二醇;巴戟天组灌胃给予5 mL浓度为1.0 mmol/L巴戟天煎剂,连续给药3个月后,体外分离培养大鼠原代破骨细胞,培养第3,6,9天进行破骨细胞TRAP染色并计数;观察两组大鼠股骨近远端骨密度值;检测两组大鼠尿Ca2+水平、血清Ca2+以及血清P水平;检测两组大鼠RANK表达情况。结果与结论:(1)破骨细胞培养第3,6,9天后,巴戟天组大鼠破骨细胞融合相对较少,并且破骨细胞融合程度减少,细胞体积相对较大,酶活性部位呈现红色。17β-雌二醇组大鼠细胞数相对较多,细胞也相对比较成熟;(2)巴戟天组大鼠左右侧股骨近、远端骨密度显著大于17β-雌二醇组(P<0.05);(3)巴戟天组尿Ca2+水平、血清Ca2+以及血清P水平,显著高于17β-雌二醇组(P<0.05);(4)巴戟天组大鼠RANK A值、RANK mR NA表达显著低于17β-雌二醇组(P<0.05)。(5)结果提示,巴戟天能降低骨质疏松大鼠骨细胞RANK表达,抑制骨质疏松的发生、发展,从而发挥对骨质疏松的保护作用。

BACKGROUND：Morinda has been reported to promote the proliferation,the secretion of alkaline phosphatase and osteocalcin,and m RNA expression of transforming growth factor of osteoblasts.However,little information is available addressing the effects of Morinda on receptor activator of nuclear factor-κB expression in osteoclasts in rats with osteoporosis.OBJECTIVE：To study the effects of Morinda on receptor activator of nuclear factor-κB expression in osteoclasts of osteoporosis rats.METHODS：Thirty Sprague-Dawley rats were equally and randomly divided into Morinda and 17β-estradiol groups.Rat models of osteoporosis were established by bilateral ovariectomy,and then 5 mL of Morinda decocta（1.0 mmol/L） and 17β-estradiol（1×10-6 mmol/L） were administered intragastrically to rats in Morinda and 17β-estradiol groups for 3 consecutive months,respectively.Primary osteoclasts were isolated from rats in both groups,and then cultured for 3,6 and 9 days followed by TRAP staining and cell counting.Bone mineral density of the proximal and distal femur,urine and serum levels of Ca2＋ and progesterone,and receptor activator of nuclear factor-κB expression in osteoclasts of rats in both groups were determined.RESULTS AND CONCLUSION：Osteoclast fusion was reduced in Morinda group.In contrast,number of osteoclasts was increased and cells became more mature in the 17β-estradiol group.Bone mineral density of the proximal and distal femur bilaterally,urine and serum levels of Ca2＋ and progesterone were significantly increased,while receptor activator of nuclear factor-κB expression was significantly decreased in osteoclasts in Morinda group compared with 17β-estradiol group（P〈0.05）.These results indicate that Morinda reduces receptor activator of nuclear factor-κB expression in osteoclasts in osteoporosis rats,thereby inhibiting the development and progression of osteoporosis.