The osteoprotective effect of Radix Dipsaci extract in ovariectomized rats

Aim

The objective of the present study was to systematically evaluate the effects of Radix Dipsaci extract (RDE) on postmenopausal osteoporosis.

Materials and methods

OVX or sham operations were performed on sixty 3-month-old virgin Sprague–Dawley rats that were divided into six groups: sham control group (sham, n = 10); OVX control group (OVX, n = 10); 17β-estradiol treatment group (E2, n = 10); three Radix Dipsaci extract treatment groups RDE100 (n = 10), RDE300 (n = 10) and RDE500 (n = 10). The treatment began 4 weeks after the surgery and lasted for 16 weeks. Bone mass, bone turnover and strength were analyzed by DEXA, biochemical markers and three-point bending test. The trabecular bone microarchitecture was evaluated by MicroCT.

Results

16 weeks treatment of RDE slowed down the body weight gain and prevented the loss of bone mass induced by the OVX. The prevention effect on bone loss was due to altering the rate of bone remodeling, which could be inferred from the decreased level of bone turnover markers, such as serum ALP, OC and urinary DPD. The changes of urinary calcium and phosphorus excretion provided the same evidence. The treatment could also enhance the bone strength and prevent the deterioration of trabecular microarchitecture.

Conclusions

Our study provides evidence that Radix Dipsaci extract will have potential to be used for treatment of postmenopausal osteoporosis.     

续断药材指纹图谱的研究

目的建立续断药材的高效液相色谱(HPLC)指纹图谱,为科学评价及有效控制续断药材质量提供可靠方法。方法采用高效液相色谱法,选用Agilent HC-C18色谱柱,流动相为乙腈(A)-(0.05mol/L)KH2PO4-H3PO4缓冲溶液pH=3.0(B),线性洗脱梯度,检测波长为220nm;流速为1.0mL/min。结果分析了10批不同时间不同产地的道地续断药材,建立了续断药材的HPLC指纹图谱,确立了17个共有峰,并且利用"中药色谱指纹图谱相似度评价软件"对不同样品之间的相似度进行计算。结论该方法准确可靠,重复性好,为续断药材的质量控制提供科学准确的依据。     

The evaluation on embryotoxicity of Dipsaci Radix with mice and embryonic stem cells

Ethnopharmacological relevance

Dipsaci Radix (DR), the dried root of Dipsacus asper Wall. ex Henry, has been used to treat pregnant disorders for thousands of years, and currently has been ranked as the first selective herb for prevention of miscarriage clinically; however, there is no sufficient evidence so far to assess its safety. The purpose of this study was to examine the impacts of DR aqueous extracts on embryonic development with mice and embryonic stem cells (ESCs).

Material and method

In a segment II study, pregnant ICR mice were randomly assigned into 5 groups, i.e. mice were orally treated with DR aqueous extracts at dosages of 0 (distilled water, as negative controls (G1 group)), 2, 8, 32 g/kg/d (G2, G3, G4 group), and vitamin A (as positive controls (G5 group)) respectively. Maternal and embryo-fetal parameters were evaluated after cesarean section. The fetal skeletal development was further assessed with the alizarin red S and H&E staining and fluorescent imaging. Meanwhile, IC₅₀ values for both ESCs and 3T3 cells were detected with MTT assays.

Result

Compared to G1 group, the maternal body-weight in G3 and G4 groups was significantly lower (P<0.05–0.001), and the fetal malformation rate increased in G2, G3 and G4 groups as a dose-dependent manner, although a statistical significance was only reached in G4 group (P<0.001). The morphologic and histochemistry abnormalities of fetal skeletal development such as delayed osteogenesis and mineralization in the cartilaginous tissue were found after DR treatments (32 g/kg/d). There was no significant difference between IC_{50 ESC} (6.826±0.311 mg/ml) and IC_{50 3T3} (5.132±0.142 mg/ml, P>0.05).

Conclusion

DR aqueous extracts at the dosage of 8 or 32 g/kg/d (4.3 or 17.2 folds of recommended daily-dosage for adult human respectively) might cause adverse impacts in maternal healthy and embryo-fetal development. It suggests that high-dose and long-term administration of DR preparations should be unsafe to pregnant women.     

续断对去势大鼠骨密度的影响

目的：探讨续断对去势大鼠骨密度的影响。方法：取10月龄雌性SD大鼠60只,随机分为正常组、模型组、雌激素组和续断低、中、高剂量组,每组10只。除正常组外,其余各组大鼠均行双侧卵巢切除术,术后1周分别给予生理盐水、续断和雌激素进行干预,共90d,观察各组大鼠全身、股骨及腰椎骨密度的变化。结果：正常组和雌激素组大鼠全身骨密度均显著高于其他各组（P〈0.05）;续断低、中、高剂量组大鼠全身骨密度与模型组比较,差异无统计学意义（P〉0.05）;续断低、中、高剂量组大鼠股骨骨密度显著高于模型组（P〈0.05）,续断中、高剂量组大鼠腰椎骨密度显著高于模型组（P〈0.05）。结论：续断能明显抑制去势大鼠股骨及腰椎骨密度的降低,对去势大鼠全身骨密度无明显影响。     

Systemic effect of crude extract from rhizome of Drynaria fortunei on bone formation in mice

The structure of the bones of mice with and without consumption of Gusuibu extracts were compared. Twenty 8-week-old male BALB/c mice were divided into two groups. In the control group ten mice were daily fed a normal diet and distilled water for drinking. In the Gusuibu group ten mice were daily fed a normal diet and distilled water mixed with Gusuibu extract for drinking. The mice were kept for 5 weeks and were then killed. Using micro-computed tomography, 20 micro-tomographic slices with an increment of 0.25 mm were acquired to cover the proximal end of the left tibia of each mouse. Quantitative morphometry of the bone structure was performed. The results showed that consumption of Gusuibu extract increased the bone volume/tissue volume ratio by 6.45%. The bone trabeculae were increased by 10.00% in number so that the bone density was increased. To conclude, Gusuibu extract taken orally increased bone density.     

生长激素联合续断对去垂体幼龄大鼠生长及骨代谢的影响

目的探究生长激素（GH）联合中药酒制续断对去垂体幼龄大鼠干预后的促生长作用及骨矿含量（BMC）的影响。方法采用经颅咽去垂体术建立生长激素缺乏大鼠模型,筛选合格动物分为假手术组（正常组,15只）、阴性对照组（模型组,13只）、GH干预组（GH组,13只）、GH联合中药续断组（联合组,12只）,GH组和联合组每日相同时间给予GH（0.25 mg/kg）颈部皮下注射,其他组注射等量生理盐水,联合组同时给予续断煎剂（0.7 mL/100 g）灌胃,其他组灌胃等量生理盐水,干预期间测量体重、尾长、身长,干预14天后采血,取股骨、胫骨,检测GH、类胰岛素生长因子I（IGF-1）、碱性磷酸酶（ALP）、骨钙素（OC）等水平,测量胫骨骨骺板宽度,用双能X线测量同侧股骨的骨密度（BMD）及BMC。结果干预2周后,与模型组比较,用药组体重、身长、胫骨长、股骨长、骨骺板宽度值、GH、IGF-1、ALP、OC升高,差异有统计学意义（P〈0.01）,尾长值增高,但差异无统计学意义（P〉0.05）,两组间以上指标比较差异无统计学意义（P〉0.05）;与模型组比较,联合组BMD值升高,差异有统计学意义（P〈0.01）。与模型组比较,用药组BMC值升高（P〈0.01）,联合组最高,差异有统计学意义（P〈0.01）。结论 GH联合中药酒制续断对促生长作用不明显,但能提高BMD和BMC,改善骨代谢。     

正交试验优选续断的水提工艺

目的:优选续断的水回流提取工艺。方法:以川续断皂苷VI转移率为评价指标,采用L9(34)正交试验考察液料比、提取时间、提取次数对续断提取工艺的影响。利用HPLC测定川续断皂苷VI含量,流动相乙腈-水(30∶70),检测波长212 nm。结果:最佳提取工艺为加15,10倍量提取2次,每次2 h;平均转移率(94.83±0.31)%。结论:该工艺简单、稳定、提取率高,适用于工业化大生产。     

Systemic effect of Fructus Psoraleae extract on bone in mice

Fructus Psoraleae extract is used in China for the treatment of bone diseases. The objective of the study was to investigate the systemic effect of Fructus Psoraleae extract consumption on bone histomorphology. Sixteen 8‐week‐old male BALB/c mice were divided into control and experimental groups. In the control group, eight mice were fed daily with distilled water. In the experimental group, eight mice were fed daily with distilled water mixed with Fructus Psoraleae extract. The mice were kept for 5 weeks and then killed. Using micro‐computed tomography, 20 micro‐tomographic slices with a separation of 0.25 mm were acquired to cover the proximal end of the left tibia of each mouse. Quantitative morphometry of the bone structure was performed. The results showed that consumption of Fructus Psoraleae extract significantly increased the bone volume/tissue volume ratio by 11.8%. The bone trabeculae increased by 7.1% in thickness so that the bone density was increased. To conclude, Fructus Psoraleae extract taken orally increases bone density and alters bone histomorphology. Copyright © 2010 John Wiley & Sons, Ltd.     

川断总皂苷对大鼠局灶性脑缺血损伤保护作用的实验研究

目的 研究川断总皂苷抗脑缺血的作用.方法 将40只大鼠随机分成为正常组、模型组、川断总皂苷低剂量组(10mg/kg)和高剂量组(50mg/kg).采用插线法阻塞大鼠大脑中动脉,制造大鼠局灶性脑缺血模型.作脑匀浆,测定脑匀浆中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性以及丙二醛(MDA)含量等指标.结果 川断皂苷低、高剂量组均能明显减少脑缺血大鼠的脑梗死面积,提高脑缺血脑中SOD、GSH-Px活性,并降低其MDA的含量.结论 川断总皂苷对大鼠局灶性脑缺血损伤有明显的保护作用.     

中药藤梨根抗肿瘤研究△

藤梨根是一种传统的清热解毒类中药,近几年关于藤梨根抗肿瘤的作用逐渐引起了人们的广泛关注,现将本课题组及国内有关专家对藤梨根的抗肿瘤作用研究近况进行综述,以期为进一步研究开发该药材提供思路。     

基于近红外光谱主成分分析-马氏距离法的发汗与未发汗续断的快速鉴别

为了快速准确的鉴别续断发汗与否, 以续断发汗和未发汗样品为实验材料, 采用近红外光谱法结合主成分分析-马氏距离判别分析方法建立了定性鉴别模型。选取了129个未发汗样品和86个发汗样品的近红外光谱图,应用主成分分析-马氏距离法进行判别分析,选择谱段为9 881.46~4 119.20 cm^-1,采用 "标准正则变换 +原始光谱+ 二阶求导" 组合对原始光谱进行预处理,主成分数为14,建立定性鉴别模型;并经预测集验证,鉴别准确率达到100%。说明近红外光谱结合模式识别方法进行续断"发汗"与否定性鉴别在技术上是可行的,可以作为续断产地加工"发汗"定性鉴别的一种辅助手段。     

续断饮片与伪品牛蒡根的FTIR区别鉴定及相关性研究

目的：为了直接准确地鉴别续断饮片及其伪品牛蒡根的真伪。方法：采用博里叶变换红外光谱法并借助OMNI采样器直接测定续断饮片及其伪品牛蒡根的红外光谱。结果：续断饮片及其伪品牛蒡根的红外光谱吸收差别较大。结论：可以采用FTIR法直接、快速、准确地对续断饮片及其伪品牛黄根进行区别鉴定。     

续断水提液对小鼠急性毒性和遗传毒性的影响

目的:探讨续断水提液对小鼠的急性毒性和遗传毒性。方法:采用最大耐受剂量法评价续断水提液对小鼠的急性毒性,观察小鼠体重及心脏、脾脏、肝脏、肾脏指数的变化;采用微核实验和单细胞凝胶电泳实验评价其遗传毒性,将40只小鼠随机分成空白组(ig 10 m L·kg~(-1)生理盐水),环磷酰胺组(第4天开始注射0.1 g·kg~(-1)环磷酰胺),续断水提液高、中、低剂量组(ig 32,16,8 g·kg~(-1)续断水提液)。各组小鼠给药5 d后处死并解剖,取骨髓细胞检测嗜多染红细胞的微核率;取胸腺、肝脏及脾脏细胞检测续断水提液对小鼠DNA的损伤。结果:续断水提液经口给药小鼠的最大给药量为80 g·kg~(-1),该剂量对小鼠体重及心脏、脾脏、肝脏、肾脏指数均无显著影响。中、低剂量组小鼠嗜多染红细胞的微核率与空白组比较无显著性差异;但续断高剂量组微核率明显高于空白组(P0.05)。高、中、低剂量组胸腺、脾脏、肝脏细胞尾部DNA百分率,尾矩略高于空白组,但均无显著性差异。结论:续断水提液对小鼠最大给药剂量为80 g·kg~(-1),属无毒级;中和低剂量续断水煎液对小鼠无遗传毒性作用,高剂量续断水提液可诱发小鼠骨髓嗜多染红细胞微核率升高,存在潜在的遗传毒性。     

星点设计-响应面法优化续断总皂苷提取工艺

目的：优选续断总皂苷的提取工艺。方法：以单因素实验为基础,选择提取浓度、提取时间和溶剂用量为自变量,以续断总皂苷含量和续断皂苷遇含量的“总评归一值”为响应值,采用星点设计-响应面法对数据进行多元线性和二项式拟合,应用响应面法优选续断总皂苷的提取工艺并对最佳工艺预测分析。结果：二项式拟合方程（R2=0.964 2）优于多元线性回归方程（R2=0.663 0）,确定最佳提取工艺为：7倍量55%乙醇回流提取3次,每次2.8 h。经最佳提取工艺提取得到续断总皂苷含量为0.201 1 g·g-1,与预测值偏差1.83%;续断皂苷遇含量为0.015 4 g·g-1,与预测值偏差3.37%。结论：星点设计-响应面法优化的提取工艺简便可行,且预测性良好,为续断总皂苷的进一步开发利用提供了依据。     

续断酒炙前后无机元素含量及其灰关联度对比分析

目的:研究续断酒炙前后样品中多种无机元素的含量,并用灰关联度法对其无机元素含量进行对比分析。方法:采用原子吸收光谱法和原子荧光光谱法测定续断片和酒续断中无机元素的含量。结果:续断酒炙前后样品中所含铁(Fe),锰(Mn),铜(Cu),锌(Zn),铅(Pb),铬(Cd),砷(As),汞(Hg)无机元素的含量各有不同,炮制后较炮制前不存在整体上升或下降趋势。结论:续断经炮制后无机元素的含量及分布发生了改变,酒炙后Fe,Mn和Zn含量增加,尤以Mn特别显著,这可能是酒炙品补肝肾的原因之一。     

续断的化学成分研究

目的研究续断的化学成分。方法运用大孔树脂、反相硅胶及制备高效液相等色谱技术进行分离纯化,并根据理化性质和波谱数据鉴定化合物的结构。结果分离得到13个化合物,其中7个环烯醚萜苷和6个木脂素类化合物,分别鉴定为马钱苷（1）、獐牙菜苷（2）、6＇-O-β-D-apiofuranosyl-sweroside（3）、续断苷H（4）、续断苷F（5）、续断苷E（6）、triplostoside A（7）、（7R,8S,7＇R,8＇S）-5-methoxyprinsepiol-4-O-β-D-glucopyranoside（8）、（7R,8S,7＇R,8＇S）-prinsepiol-4-O-β-D-glucopyranoside（9）、acanthoside D（10）、（7R,8S,7＇R,8＇S）-fraxiresinol-4＇-O-β-D-glucopyranoside（11）、（7R,8S,7＇R,8＇S）-8-hydroxypinoresinol-4＇-O-β-D-glucopyranoside（12）、（7R,8S,7＇R,8＇S）-8-hydroxypinoresinol-4-O-β-D-glucopyranoside（13）。结论其中化合物8、9、11、12、13为首次从川续断属植物中分离得到。     

中药四性的生物热动力学研究 --人参和西洋参药性的微量量热学比较

目的：从生物物理化学的角度,阐明中药四性（寒、热、温、凉）的客观真实性。方法：利用微量量热法,测定了大肠杆菌在不同条件下的生长热谱曲线,得到了相应的生物热动力学参数,如生长速率常数（K）、抑制率（Ⅰ）,半抑制率（IC50）、热焓（△H）。根据生物热动力学参数值,结合本草文献报道,综合分析了人参和西洋参药性在。结果;人参和西洋参抑制细菌生长的热谱曲线,其形状基本相同,随着药物浓度的加大,代谢过程的停滞期延长,生长代谢峰后移;其热力学参数热焓（△H）存在较稳定的差异－－人参和西洋参均降低了细菌生长代谢过程中热量的释放。但相同浓度下,人参降低的幅度大于西洋参。结论：微量量热法可作为刻划中药药性的一个新的有效手段,亦可作为中草药活性筛选的新手段。     

基于UPLC-QTOF-MS/MS和TCMIP的续断治疗类风湿性关节炎活性成分筛选及作用机制分析

目的:依托超高效液相色谱-四极杆飞行时间质谱法(UPLC-QTOF-MS/MS)和中医药整合药理学研究平台(TCMIP),探讨续断治疗类风湿性关节炎(RA)的主要活性成分和作用机制.方法:采用UPLC-QTOF-MS/MS,分别在正、负离子扫描模式下对续断醇提液中的化学成分进行定性分析,流动相0.1％甲酸水溶液(A)-...     

红芪对血液流变性的影响

目的：研究红芪对血液流变性的影响。方法：采用正常、寒凝血瘀(肾上腺素 冰水应激)、气虚血瘀(灌服番泻叶)模型大鼠,观察红芪醇提物和水提物对血液粘度、血沉、红细胞压积、电泳时间等血液流变学指标影响,并考察其对ADP诱导家兔体外血小板聚集的影响。结果：红芪醇提物可明显降低正常大鼠高切(80s^-1)和低切(20s^-1)下全血比粘度,红芪水提物可明显减轻正常大鼠体外血栓的干重,显降低肾上腺素加冰水浴所致血瘀模型大鼠体外血栓湿重及干重,对血栓长度、全血比粘度及血沉等指标有一定抑制趋势;二均可明显降低番泻叶所致气虚血瘀模型大鼠红细胞压积。缩短其红细胞电泳速率;体外125、250、500μm／m1可剂量依赖性地抑制ADP引起的家兔血小板聚集。结论：红芪具有一定活血化瘀作用。     

刺五加提取物对实验性急性脑缺血的保护作用

目的 观察刺五加提取物对急性脑缺血大鼠和沙鼠脑毛细血管通透性、脑指数及脑含水量的影响。方法 用结扎双侧颈总动脉法建立动物急性脑缺血模型，股静脉iv伊文思蓝和测脑组织干重的方法观察刺五加提取物对急性脑缺血大鼠和沙鼠脑毛细血管通透性、脑指数及脑含水量的影响。结果 刺五加提取物对急性脑缺血大鼠和沙鼠脑毛细血管通透性、脑指数及脑含水量都有非常明显的降低作用。结论 刺五加提取物对实验性脑缺血有保护作用，且明显优于刺五加注射液。