“补肾活血方”对绝经后骨质疏松症影响的实验研究

发布时间：2018-03-21 12:46

本文选题：补肾活血方　切入点：绝经后骨质疏松症　出处：《南京中医药大学》2016年博士论文　论文类型：学位论文

【摘要】：目的：研究江苏省中医院骨伤科经验方“补肾活血方”对绝经后骨质疏松症的疗效机制。方法：分别基于“补肾活血”方药治疗骨质疏松症随机对照临床试验文献的系统评价和Meta分析；“补肾活血方”对去卵巢骨质疏松症大鼠模型“高转换型”骨代谢失衡的调控作用；“补肾活血方”含药血清对人成骨细胞增殖、分化和矿化能力的干预作用三个方面,探索“补肾活血”治疗绝经后骨质疏松症的疗效机制。①文献系统评价和Meta分析：关键词检索(补肾、补肝肾、活血、健脾活血、骨质疏松症、临床试验、临床研究),检索数据库为维普、万方、中国知网,纳入2000年01月-2016年9月时间区间内以补肾活血为法则的方药治疗骨质疏松症的随机对照临床试验。应用Review Manager 5.3软件进行Meta分析,所选用主要结局指标为：主观症状改善、骨密度改变、腰背痛VAS评分改善。通过Meta分析数据说明“补肾活血”方药治疗骨质疏松症确有临床疗效；并基于Jadad文献质量评分体系,分析临床研究存在的不足。②“补肾活血方”对造模大鼠“高转换型”骨代谢失衡的调控作用：设立四组动物(空白对照组、模型对照组、治疗组、假治疗组),每组25只。造模方法为人工手术去除大鼠双侧卵巢制作骨质疏松症动物模型,治疗组造模2周后开始给药(补肾活血汤浓缩液灌胃),假治疗组给予生理盐水灌胃。分别于4周、8周、12周每组取5只大鼠经中心静脉取血检测骨代谢相关指标：抗酒石酸酸性磷酸酶5b(TRACP-5b)、骨碱性磷酸酶(BALP)、I型前胶原N端前肽(PINP)、I型前胶原羧基端肽(PICP)、I型胶原C端肽(CTX-i)、I型胶原N末端肽(NTX)；第12周中心静脉取血后麻醉各组大鼠,检测L2-L6椎体骨密度和骨矿含量。③“补肾活血方”含药血清对人成骨细胞增殖、分化和矿化能力的干预作用：取骨质疏松症合并股骨颈骨折患者松质骨进行成骨细胞培养,构建成骨细胞分化模型：制备低、中、高三种浓度的补肾活血方含药血清；采用MTT法、细胞上清液碱性磷酸酶测定和茜素红染色观察含药血清对成骨细胞的影响。结果：①文献系统评价和Meta分析：结果显示“补肾活血”方药治疗骨质疏松症在主观症状改善、骨密度提高、疼痛VAS评分改善较对照治疗存在优效性,但此类临床研究的随机化方案设计、盲法实施等科研方案设计方面存在不规范性,需要后续更严谨的大样本、多中心、随机对照临床试验证据的进一步支持；②动物实验：造模后4W模型对照组、治疗组、假治疗组大鼠血清BALP、CTX、 NTX和TRACP含量显著高于空白对照组；造模后8周和12周,模型对照组、假治疗组上述指标的增高趋势仍然维持,而治疗组则在第8周和12周出现较模型对照组和假治疗组下降的趋势。模型对照组、治疗组、假治疗组大鼠血清PICP和PINP水平则在造模后4周较空白对照组显著下降,造模后8周和12周模型对照组和假治疗组的PICP和PINP水平仍然呈下降趋势,而治疗组大鼠的血清PICP和PINP水平则在造模后第8W即出现回升,造模后12W也保持了增高的趋势,但含量较空白对照组仍较低。③细胞实验：加入含药血清干预细胞生长48h后,MTT检测显示中、高剂量组出现不同程度的细胞增殖现象,并存在一定程度的剂量依赖；作用72h后,低、中、高三个剂量组ALP活性增高,高剂量组茜素红染色钙盐沉积明显增多。结论：江苏省中医院骨伤科经验方“补肾活血方”对绝经后骨质疏松症的疗效与以下作用机制有关：①对绝经后骨质疏松症“高转换型”骨代谢失衡紊乱的调节,即对异常增高的骨转换速率起到了抑制性作用；②有提高人成骨细胞内ALP活性和促进成骨细胞增殖、分化及矿化的作用。
[Abstract]:Objective: To study the Jiangsu Province Traditional Chinese Medicine Hospital orthopedic experience of "Bushen Huoxue Decoction" on the therapeutic mechanism of postmenopausal osteoporosis. Methods: Based on "Bushen Huoxue Prescription" treatment of osteoporosis in randomized controlled clinical trial literature system evaluation and Meta analysis; the regulation effect of tonifying kidney Activating Blood Prescription on rat model of "imbalance ovarian osteoporosis high conversion" bone metabolism; "Bushen Huoxue Decoction" on serum osteoblast proliferation, three aspects of the intervention effect of differentiation and mineralization, exploration of "Bushen Huoxue" in the treatment of postmenopausal osteoporosis. The therapeutic mechanism of evaluation and analysis of Meta document system: keyword search (kidney, liver and kidney, promoting blood circulation, invigorating spleen and promoting blood circulation, osteoporosis, clinical trials, clinical research, database retrieval) for VIP, Wanfang, China HowNet, in 2000 01 months -2016 year in September the time zone In the kidney and activating blood in the treatment of osteoporosis is the law of prescription of randomized controlled clinical trials. The application of Review Manager 5.3 Meta analysis software, the main outcome measures were: subjective symptoms, bone mineral density, low back pain VAS score improvement. Through the analysis of Meta data shows that "Bushen Huoxue Prescription" treatment of osteoporosis osteoporosis is clinical curative effect; and based on Jadad document quality scoring system, analysis of the lack of clinical research there. The regulation of "Bushen Huoxue Decoction" on the imbalance of rats "high conversion type" bone metabolism: set up four groups of animals (blank control group, model control group, treatment group, sham treatment group), 25 rats in each group. The modeling method for the artificial surgical removal of both ovaries of rats produced osteoporosis animal model, rats in the treatment group 2 weeks after administration (BSHX concentrated liquid gavage), sham treatment group received saline Saline. At 4 week, 8 weeks, 12 weeks, 5 rats from each group by central venous blood bone metabolism index detection: tartrate resistant acid phosphatase 5b (TRACP-5b), bone alkaline phosphatase (BALP), type I procollagen N peptide (PINP), before the end of precollagen I carboxy terminal peptide (PICP), type I collagen C terminal peptide (CTX-i), type I collagen N terminal peptide (NTX); central venous blood after twelfth weeks all rats were anaesthetized, detection of L2-L6 vertebral bone mineral density and bone mineral content. The "Bushen Huoxue Decoction" on serum osteoblast proliferation, intervention of differentiation and mineralization ability: osteoporosis complicated with femoral neck cancellous bone osteoblast culture fracture patients, construct osteoblast differentiation model: preparation of low, high concentration of Bushenhuoxue Decoction medicated serum by MTT method; cell supernatant alkaline phosphatase staining and alizarin red medicated serum on osteoblasts The influence of cells. Results: Meta analysis and Literature: the results show that the system of "Bushen Huoxue Decoction" in the treatment of osteoporosis disease in subjective symptoms, increase bone density, pain VAS score better than the control treatment are effective, but the clinical research of such randomized scheme design, there is no normative aspects of design for the blind the implementation of the law and other research programs need more rigorous follow-up of large sample, multicenter, randomized controlled clinical trials to further support the evidence; the animal experiment: after modeling, 4W model control group, treatment group, serum BALP group, sham treatment of CTX, NTX and TRACP was significantly higher than the control group after modeling; 8 and 12 weeks, the model control group, sham treatment group increased the index remained unchanged, while the treatment group in eighth weeks and 12 weeks compared with model control group and sham treatment group decreased. The model control group, sham treatment group. The treatment of serum PICP and PINP levels of rats in 4 weeks after surgery than the control group significantly decreased at 8 weeks and 12 weeks model group and sham treatment group of PICP and PINP levels are decreased, while the treatment group rats serum PICP and PINP levels in the model the 8W is a rebound after modeling 12W also maintained a rising trend, but the content is still lower than the control group. The cells in the experimental intervention: serum containing 48h cell growth after MTT assay, cell proliferation phenomenon was found in different degree in high dose group, and there is a certain degree of dose dependence; 72h, low, high dose group increased the activity of ALP, the high dose group of alizarin red staining calcium deposition increased significantly. Conclusion: the Jiangsu Province Traditional Chinese Medicine Hospital orthopedic experience of "Bushen Huoxue Decoction" on the treatment of postmenopausal osteoporosis and the mechanism: (1) Regulation of the "high transformation" bone metabolic disturbance disorder in postmenopausal osteoporosis, that is, it plays an inhibitory role in the abnormal increase of bone turnover rate. It has the effect of enhancing ALP activity and promoting osteoblast proliferation, differentiation and mineralization in human osteoblasts.

【学位授予单位】：南京中医药大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R259

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