Protective Effects of Total Flavonoids from Epimedium on the Male Mouse Reproductive System Against Cyclophosphamide‐Induced Oxidative Injury by Up‐Regulating the Expressions of SOD3 and GPX1

Total flavonoids of Epimedium (TFE) is the main active composition of Epimedium that has been used to treat male reproductive problems. The present aim was to investigate the protective effects of TFE on male mice reproductive system against cyclophosphamide (CP)‐induced oxidative injury. The animals were treated with CP to make testicular injury model and the protective effects of TFE were observed. In the CP‐treated group, testicular and epididymal weights, sperm count and motility significantly decreased relative to the control group (P < 0.05 and P < 0.01, respectively). Compared with the CP‐treated group, TFE (200 and 400 mg/kg) treated mice increased testicular weights by 21.6% and 28.4% (P < 0.05), sperm counts by 81.7% and 148.3% (P < 0.01) and sperm motility by 47.2% and 61.3% (P < 0.01). Meanwhile, the CP‐treated group showed enhancement of lipid peroxidation leading to testicular reproductive toxicity. TFE restored these oxidative damages by up‐regulating the expression of antioxidant enzymes, especially SOD3 and GPX1. TUNEL assay and histopathological observations provided supportive evidence for above results, and when the dose of TFE increased, the aforesaid improvement became more and more strong. These results demonstrated that TFE exerted beneficially protective effects on the structural and functional damage of male mice reproductive system and reduced apoptosis in spermatogenic cells by inhibiting CP‐induced oxidative stress. Copyright © 2013 John Wiley & Sons, Ltd.     

淫羊藿苷促进胶原水凝胶三维立体培养成骨细胞的成熟分化

目的研究淫羊藿苷对胶原水凝胶三维立体培养的新生大鼠颅骨成骨细胞(ROB)分化和成熟的影响。方法取新生SD大鼠颅骨, 酶消化法培养ROB, 将传代1次的ROB包埋于2 mg/mL的Ⅰ型鼠尾胶原中进行三维立体培养, 48 h后进行双乙酸荧光素/碘化丙啶(FDA/PI)细胞染色、苏木素-伊红(HE)染色, 并用扫描电镜观察细胞生长状态。分别采用终浓度为1×10 ^-4、1×10 ^-5、1×10 ^-6和1×10 ^-7 mol/L的淫羊藿苷进行干预, 培养3、6、9 d后分别检测细胞内碱性磷酸酶活性, 筛选最佳药物浓度; 以最佳浓度淫羊藿苷干预三维凝胶中的ROB 12、24、36、48 h后, 用实时定量PCR和蛋白质印迹法分别检测骨形态发生蛋白2(BMP-2)、Runt相关转录因子2(RUNX-2)、Osterix mRNA及其蛋白表达情况。 结果胶原水凝胶三维立体培养的ROB采用FDA/PI染色、HE染色以及扫描电镜结构显示, ROB在胶原水凝胶中分布均匀, 生长状态良好。终浓度为1×10 ^-5、1×10 ^-6、1×10 ^-7 mol/L的淫羊藿苷可显著提高三维凝胶中ROB的碱性磷酸酶活性, 其中以1×10 ^-6 mol/L浓度淫羊藿苷干预后ROB的碱性磷酸酶活性最高。1×10 ^-6 mol/L淫羊藿苷能显著提高BMP-2、RUNX-2、Osterix mRNA及其蛋白表达量。 结论淫羊藿苷可显著促进三维立体培养的ROB成骨性相关因子的表达, 提示淫羊藿苷对胶原水凝胶三维立体培养的ROB具有较强的促骨形成活性。     

淫羊藿、肉蓉、巴戟天对果蝇寿命影响的研究

目的研究淫羊藿、肉蓉、巴戟天3种药物对果蝇寿命的影响。方法采用在实验组的培养基中适量加入上述的3种药物,而在对照组的培养基中则不加药物,观察各组的生存时间。结果实验组果蝇的平均寿命高于对照组果蝇的平均寿命。结论适量应用淫羊藿、肉蓉、巴戟天,具有延缓衰老的功效。     

高效液相色谱法测定骨疏灵颗粒中淫羊藿苷含量

目的建立HPLC法测定骨疏灵颗粒中淫羊藿苷的含量。方法采用Cosmosil C18(250 mm×4.6 mm,5μm)为色谱柱,流动相:乙腈-水(30∶70),流速:1.0 m L/min,检测波长:270 nm。结果淫羊藿苷在0.01~0.25 mg/m L范围内呈良好的线性关系(r=0.999 9),回收率为99.99%(RSD=0.75%,n=6)。结论本方法简单、准确,灵敏度高,重现性好,可用于骨疏灵颗粒中淫羊藿有效成分淫羊藿苷的含量测定。     

藿草壮骨合剂中淫羊藿和熟地黄的薄层色谱鉴别

目的：建立藿草壮骨合剂的质量控制方法。方法采用薄层色谱法对处方中淫羊藿、熟地黄进行鉴别。结果在薄层色谱上鉴别出制剂中淫羊藿、熟地黄的特征斑点,且斑点清晰,无干扰。结论该法结果可靠,方法简便,专属性强,重现性、耐用性好,可为藿草壮骨合剂的质量控制提供参考。     

Efficient removal of the organochlorine pesticide and heavy-metal residues in<Emphasis Type="Italic">Epimedium brevicornum Maxim</Emphasis> by supercritical fluid extraction

A method involving depuration of 12 organochlorine pesticides (OCPs) and 7 heavy metals from Epimedium brevicomum Maxim was developed using supercritical fluid extraction (SPE). The pesticides in the study consisted of alpha, beta-, gamma-, and delta-Benzene hexachloride, Pentachloronitrobenzene (PCNB), Pentachloroaniline (PCA), Heptachlor (HEPT), Methyl -pentachlorophenyl sulfide (MPCPS), pp'-DDE[1,1-dichloro-2, 2-bis (p-chlorophenyl) ethylene], op'-DDT [1,1,1-trichloro-2-(o-chlorophenyl)-2-(p-chlorophenyl)ethane], pp'-DDD [1,1-dichloro-2-2-bis(p-chlorophenyl)]ethane, pp'-DDT [1,1,1-trichloro-2,2-bis(p-chlorophenyl) ethane]. A series of experiments was conducted to optimize the final extraction conditions as following: pure CO2, extraction pressure of 15 Mpa, extration temperature of 60 degrees C, extraction time of 10 min, flow rate at 55 kg/h. A GC method with electron capture detection was employed to determination of the OCPs, and an atomic absorption spectrometry (AAS) was designed for the determination of 7 heavy metals including of Pb, Cd, Cu, Fe, Zn, As, Hg in Epimedium brevicomum Maxim. A HPLC method was developed for the quantitative determination of active constituents. The SFE was used to remove the organochlorine pesticide and heavy metals from Epimedium brevicornum Maxim, receiving high decontamination rate of pesticide residue and low loss of active constituents.     

Effect of lipid-based suspension of Epimedium koreanum Nakai extract on sexual behavior in rats

Ethnopharmacological relevance

Herba of Epimedium koreanum is used in traditional Chinese and Korean herbal medicine as a potent enhancer of erectile function. Icariin, the main active component of Epimedium koreanum, possesses many biological effects, such as improving cardiovascular function, hormone regulation, immunological function modulation, and anti-tumor activity.

Aim of the Study

This study supports the traditional use of extracts from Epimedium species in erectile dysfunction.

Materials and Methods

The Epimedium koreanum dry extract was suspended in wheat germ oil using lecithin and bee wax for oral administration. The effect of oral administration of two compositions (E-01 and E-02) standardized by their icariin content on the number of complete intromissions, the number of ejaculations, and the latent period of ejaculation (LPE) in rats were evaluated. E-01 and E-02 were administered orally for 10 days to the experimental animals. The control animals received olive oil for 10 days. On day 10, 0.5 h after the dose was administered to male rats, one virgin female rat was placed with one male rat.

Results

The number of complete intromissions increased to 23.3 ± 2.6 in the E-01 and E-02 group (dose 300 mg/kg body weight) (b.wt) and to 20.1 ± 2.3 in the E-02 group (dose 750 mg/kg b.wt) compared with 15.2 ± 2.4 in the control group of aged rats. The number of ejaculations increased from 1.1 ± 0.3 in the control-aged group to 2.6 ± 0.4 in the E-01 group. The LPE of male rats was 14.2 ± 1.8 min in the control-aged group. The LPE of the aged group was reduced to 9.8 ± 1.5 min, 9.8 ± 1.6 min, and 11.4 ± 1.8 min when treated with E-01 at a dose of 300 mg/kg b.wt, and E-02 at a dose of 300 mg/kg b.wt and 750 mg/kg b.wt, respectively.

Conclusion

It was established that oral administration of lipid-based suspension of dry extract of Epimedium koreanum in wheat germ oil improved erectile function of aged rats.     

淫羊藿药材的指纹图谱

目的:建立淫羊藿药材的高效液相指纹图谱分析方法。方法:采用Agilent ODS-C18(4.6mm×150mm,5μm)色谱柱,柱温:40℃,流速:1mL/min。以乙腈、水为流动相进行梯度洗脱,检测波长为270nm;以淫羊藿苷为参照物。结果:通过对8个不同产地的淫羊藿药材样品的测定,标定了10个共有峰特征,并通过"中国色谱指纹图谱相似度评价系统2004A版软件",计算出其相似度。结论:该方法可为更好的控制淫羊藿药材的内在质量提供科学依据。     

“抗衰复春片”质量标准的研究

目的：研究抗衰复春片的质量标准。方法：增加TLC法鉴别何首乌、五味子、淫羊藿及RP-HPLC法测定橙皮苷的含量。含量采用Waters C18色谱柱（4.6mm×200mm,5μm）;流动相为甲醇-0.1%磷酸溶液（35∶65）,流速1mL·min-1;检测波长：283nm;用外标法定量。结果：鉴别：阴性对照均无干扰;含量：0.2084～4.1680μg范围内有良好的线性关系（r=0.9999）,平均加样回收率为98.4%（n=6）。结论：所建方法简单、可靠,可用于抗衰复春片的质量控制。     

淫羊藿属植物PCR-RFLP遗传多样性研究

目的建立淫羊藿植物PCR-RFLP标记系统关系图。方法对主要来自于四川、贵州的17种淫羊藿植物进行了系统的PCR-RFLP标记。结果在8个引物的PCR-RFLP标记分析中,有7个标记能得到1条清晰的谱带;利用12种限制性内切酶对7个标记的扩增产物消化后,在84种引物/酶组合中,共检测到129条酶切片段,其中44条具有多态性;遗传相似系数为0.550～0.988,平均为0.821。结论该属植物遗传关系与其地理分布关系密切。