健骨颗粒对去卵巢骨质疏松模型鼠血清肿坏死因子α及转化生长因子β1的影响

背景在前期实验的基础上,本实验进一步研究健骨颗粒对去卵巢骨质疏松模型鼠血清肿瘤坏死因子α、转化生长因子β1含量的变化及相关性,借以深入探讨其对骨代谢的调节作用.目的观察去卵巢骨质疏松模型鼠血清中肿瘤坏死因子α、转化生长因子β1含量变化以及相关性,分析补肾健脾中药健骨颗粒对去卵巢骨质疏松模型鼠骨代谢的具体调节机制.设计以实验动物为观察对象,通过摸球法进行随机化分组.单位福建中医学院骨伤系、福建中医学院中心实验室.对象对象的样本量为6月龄SD雌性大鼠50只,体重350～400 g,随机分成5组,每组10只,以切除卵巢为模型,用单纯随机化方法分成5组,并规定第1组为假手术组,即行腹部切口但不切除卵巢.第2～5组为造模组,即进行腹部切口双侧卵巢切除术造模.其中第2组为单纯模型组;第3组为生理盐水组;第4组为骨松宝组(骨松宝成份淫羊藿、川芎、牡蛎等);第5组为健骨颗粒组(健骨颗粒成份煅狗骨、淫羊藿、党参等中药).方法假手术组只行腹部切口但不切除卵巢,将第2～5组切除卵巢造模.于术后3个月时腹腔注射麻醉药40mg/kg戊巴比妥处死假手术组和单纯模型组大鼠,经相关指标检测确定模型成立.第3～5组大鼠按照组别分别喂服生理盐水2 mL/(kg·d)、骨松宝1.5 g/(kg·d)和健骨颗粒2g/(kg·d),统一于术后第13周,即模型成立后给药,用药12周后在麻醉状态下统一处死.取颈动脉血检测血清中肿瘤坏死因子-α、转化生长因子-β1及雌二醇的含量,结果用x±s表示,组间比较用t检验.主要观察指标去卵巢后12周及喂服药物12周后各组大鼠血清肿瘤坏死因子α、转化生长因子β1、雌二醇的含量以及其相关性.结果①去卵巢后12周,随着雌二醇水平下降,血清肿瘤坏死因子α含量明显升高,而转化生长因子β1浓度却明显降低.②当喂服健骨颗粒12周后,血清雌二醇和转化生长因子β1浓度(54.05±15.92),(9.12±1.79)ng/L]回升明显,与同期生理盐水组相比(27.24±2.99),(5.53±1.46)ng/L]差异有显著性意义(t=5.234,t=4.907,P＜0.01),而肿瘤坏死因子α含量与生理盐水组相比却出现下降(32.44±3.42),(65.86±35.36)ng/L,t=2.975,P＜0.05].③血清肿瘤坏死因子α与雌二醇含量呈负相关(r=-0.324,P＜0.05);血清转化生长因子β1与雌二醇含量呈高度正相关(r=0.673,P＜0.01);血清肿瘤坏死因子α与转化生长因子β1亦呈负相关(r=-0.387,P＜0.02).结论健骨颗粒通过调整骨质疏松模型鼠体内雌二醇水平,制约肿瘤坏死因子α的合成、分泌及其活性,解除对转化生长因子β1的抑制作用,使破骨细胞的骨吸收活动减弱,成骨细胞的骨形成作用增强,改善骨代谢的负平衡.

Effect of Jiangu granules on serum tumor necrosis factor-α and transforming growth factor-β1 in osteoporotic rats with ovariectomy

BACKGROUND: On the basis of the results of previous experiments, this experiment further examined the changes in serum tumor necrosis factor- α(TNF-α) and transforming growth factor-β1 (TGF-β1) in osteoporotic rats treated with Jiangu granules, and analyzed the correlation between the changes and osteoporosis to evaluate the effect of Jiangu granules in bone metabolism regulation.OBJECTIVE:To observe the changes in serum TNF-α and TGF-β1 in rat models of osteoporosis induced by ovariectomy for analysis of the mechanism by which Jiangu granules, a kidney-tonifying and spleen-invigorating traditional Chinese medicine preparation, regulates bone metabolism.DESIGN: Randomized controlled experiment.SETTING:Department of Orthopedic Injury and Central Laboratory of Fujian College of Traditional Chinese Medicine.MATERIALS :Totally 50 6-month-old female SD rats weighing 350-400 g were randomized equally into 5 groups. One group served as the sham operation group, in which the rats were only subjected to abdominal incision without ovariectomy, and the rats in the other 4 groups received bilateral ovariectmomy to establish rat models of osteoporosis. The latter 4 groups received treatment with normal saline, Gusongbao (a tradition Chinese medicine preparation composed of Epimedium sagittatum, Rhizoma Chuanxiong and oyster shell etc.), Jiangu granules (composed of calcined dog bone, Epimedium sagittatum, and Codonopsis pilosula, etc.), or no treatment (model group).METHODS :Three months after model establishment, the rats in sham operation and simple model groups were anaesthetized by intraperitoneal injection of 40 mg/kg pentobarbital for confirmation of successful model establishment by examination of relevant indexes. From the 13th week after operation, the rats in the other 3 groups were respectively given saline (2 mL/kg), Gusongbao (1.5 g/kg) and Jiangu granules (2 g/kg) on a daily basis for 12 weeks. After the completion of the treatment course, the rats were anaesthetized and the carotid artery blood was taken for the examination of serum TNF-α, TGF-β1 and estradiol contents.MAIN OUTCOME MEASURES:Serum levels of TNF-α, TGF-β1 and estradiol 12 weeks after ovariectomy and 12 weeks after drug administration and their correlation.Treatment with Jiangu granules for 12 weeks resulted in obvious elevation of serum estradiol and TGF-β1 levels (54.05±15.92) ng/L and (9.12±1.79) ng/L,respectively] and reduction of TNF- α (32.44±3.42) ng/L] in comparison with those in saline-treated rats (27.24±2.99) ng/L, (5.53±1.46) ng/L, and (65.86±35.36) ng/L, t=5.234, t=4.907, P ＜ 0.01 for estradiol and TGF-β1was inversely correlated with estradiol (r=-0.324, P ＜ 0.05) and TGF-β1was highly positively correlated with estradiol (r =0. 673, P ＜ 0. 01). Serum TNF-α was also inversely correlated with TGF-β1 (r=-0.387, P＜ 0. 02).CONCLUSION:Jiangu granules modulates the level of estradiol in osteoporotic rats to suppress the synthesis, secretion and activation of TNF-α,alleviate the inhibition of TGF-β1, attenuate the bone-absorption activity of osteobalsts and enhance the osteogenic activity of osteoplasts, thereby correct the negative balance of bone metabolism.