Aqueous extract of Epimedium sagittatum mitigates pulmonary fibrosis in mice / 中国中药杂志

This study aims to investigate the intervention effect of the aqueous extract of Epimedium sagittatum Maxim on the mouse model of bleomycin(BLM)-induced pulmonary fibrosis, so as to provide data support for the clinical treatment of pulmonary fibrosis. Ninety male C57BL/6N mice were randomized into normal(n=10), model(BLM, n=20), pirfenidone(PFD, 270 mg·kg~(-1), n=15), and low-, medium-, and high-dose E. sagittatum extract(1.67 g·kg~(-1), n=15; 3.33 g·kg~(-1), n=15; 6.67 g·kg~(-1), n=15) groups. The model of pulmonary fibrosis was established by intratracheal instillation of BLM(5 mg·kg~(-1)) in the other five groups except the normal group, which was treated with an equal amount of normal saline. On the day following the modeling, each group was treated with the corresponding drug by gavage for 21 days. During this period, the survival rate of the mice was counted. After gavage, the lung index was calculated, and the morphology and collagen deposition of the lung tissue were observed by hematoxylin-eosin(HE) and Masson staining, respectively. The levels of reactive oxygen species(ROS) in lung cell suspensions were measured by flow cytometry. The levels of glutathione peroxidase(GSH-Px), total superoxide dismutase(T-SOD), and malondialdehyde(MDA) the in lung tissue were measured. Terminal-deoxynucleoitidyl transferase-mediated nick-end labeling(TUNEL) was employed to examine the apoptosis of lung tissue cells. The content of interleukin-6(IL-6), chemokine C-C motif ligand 2(CCL-2), matrix metalloproteinase-8(MMP-8), transforming growth factor-beta 1(TGF-β1), alpha-smooth muscle actin(α-SMA), E-cadherin, collagen Ⅰ, and fibronectin in the lung tissue was measured by enzyme-linked immunosorbent assay(ELISA). The expression levels of F4/80, Ly-6G, TGF-β1, and collagen Ⅰ in the lung tissue were determined by immunohistochemistry. The mRNA levels of CCL-2, IL-6, and MMP-7 in the lung tissue were determined by qRT-PCR. The content of hydroxyproline(HYP) in the lung tissue was determined by alkaline hydrolysation. The expression of α-SMA and E-cadherin was detected by immunofluorescence, and the protein levels of α-SMA, vimentin, E-cadherin in the lung tissue were determined by Western blot. The results showed the aqueous extract of E. sagittatum increased the survival rate, decreased the lung index, alleviated the pathological injury, collagen deposition, and oxidative stress in the lung tissue, and reduced the apoptotic cells. Furthermore, the aqueous extract of E. sagittatum down-regulated the protein levels of F4/80 and Ly-6G and the mRNA levels of CCL-2, IL-6, and MMP-7 in the lung tissue, reduced the content of IL-6, CCL-2, and MMP-8 in the alveolar lavage fluid. In addition, it lowered the levels of HYP, TGF-β1, α-SMA, collagen Ⅰ, fibronectin, and vimentin, and elevated the levels of E-cadherin in the lung tissue. The aqueous extract of E. sagittatum can inhibit collagen deposition, alleviate oxidative stress, and reduce inflammatory response by regulating the expression of the molecules associated with epithelial-mesenchymal transition, thus alleviating the symptoms of bleomycin-induced pulmonary fibrosis in mice.

A new flavonoid glycoside from Epimedium sagittatum / 药学学报

Fourteen flavonoids were isolated and purified from Epimedium sagittatum by various chromatography techniques such as macroporous adsorbent resin, silica gel, ODS, Sephadex LH-20, HW-40C and semi-preparative HPLC. Their structures were identified by analysis of physicochemical properties and spectral data, and determined as 3′-hydroxy-baohuoside-Ⅱ (1), huazhongilexone-7-O-β-D-glucopyranoside (2), kaempferol-3-O-α-L-rhamnoside (3), baohuoside-Ⅱ (4), icariside-Ⅱ (5), kaempferol 3,7-di-O-α-L-rhamnopyranoside (6), (+)-aromadendrin (7), kaempferol 3-O-(2-O-β-D-apiofuranosyl)-α-L-rhamnopyranoside (8), sagittatoside A (9), 2″-O-rhamnosyl icariside-II (10), apigenin-7-O-β-D-glucoside (11), quercetin 3-O-β-D-apiofuranoyl-(1→2)-α-L-rhamnopyranoside (12), kaempferol (13), icariin (14). Among them, compound 1 is a new compound, while compounds 2, 6-8, 11, and 12 were isolated from E.sagittatum for the first time.

Distribution of Epimedium sagittatum and its research progress on quality characteristics / 中草药

To promote the rational development and utilization of Epimedium sagittatum, and to propose the utilization strategies and future research suggestions. The distribution was illuminated based on the examination of herbarium specimens and our field investigation. At the same time, the quality characteristics of different harvest time, different parts and different producing areas were summarized according to the literature review and our previous researches. In terms of the distribution, E. sagittatum is a widely distributed species in Epimedium. However, there were many errors in the identification of herbarium specimens. At least 13 taxa were wrongly identified as E. sagittatum. The distribution of E. sagittatum was not as extensive as previously recorded in the literature. Although the distribution of E. sagittatum has been recorded in 12 provinces and in Chongqing Municipality, it was mainly distributed in Anhui, Jiangxi, Hunan and Hubei provinces, and the resources in each region were not abundant. In terms of quality characteristics, there were great differences in both composition and contents of the active components among different germplasm. Some of the germplasm in Hunan, Hubei and Sichuan provinces were excellent. However, the stability and homogeneity were not ideal. Therefore, the utilization of E. sagittatum should depend on the quality characteristics. It was found that there were great differences among populations and which were closely related to the genetic basis. The effect of physiological ecology on quality and its mechanism, as well as the molecular mechanism of quality formation should be paid more attention in the future.

Evaluation of metabolism-effect/toxicity of Epimedium sagittatum based on zebrafish M-Act/Tox integration method / 中草药

Objective To evaluate the metabolism, anti-osteoporosis efficacy and toxicity of Epimedium sagittatum based on zebrafish M-Act/Tox integration method. Methods The safety of E. sagittatum (crude drug 200, 500, 1 000, 1 500, and 2 000 μg/mL) was evaluated with 1-6 dpf zebrafish, the morphology of fish organs was observed and the number of deaths was counted and the half death concentration of zebrafish (LC50) was calculated; The 5 dpf zebrafish were exposed to E. sagittatum (crude drug 250 μg/mL ) for 24 h, six representative flavonoids, including epimedin A, epimedin B, epimedin C, icariin, sagittatoside C, and baohuoside I, were analyzed; The zebrafish osteoporosis model was induced with 25 μmol/L prednisolone, microscopic detection and digital imaging of zebrafish larvae of each group cultured to 8 dpf were performed using alizarin red, and the bone staining area was quantitatively analyzed by image software to evaluate the anti-osteoporosis activity of E. sagittatum. Results E. sagittatum caused zebrafish poisoning at crude drug 500 μg/mL and above concentration (organs deformation or death), and toxicity was related to drug concentration and exposing time; After the action of zebrafish, epimedin A, epimedin B, epimedin C, and icariin were almost completely transformed, and sagittatoside C and baohuoside I were the main metabolites; E. sagittatum had significant anti-prednisolone-induced bone loss in zebrafish at a certain concentration (crude drug 6.25, 12.5, and 25 μg/mL). Conclusion The metabolism, anti-osteoporosis efficacy and toxicity of E. sagittatum are evaluated using zebrafish. The zebrafish M-Act/Tox integration method integrates the advantages of zebrafish metabolism, osteoporosis model and toxicity evaluation method, and realizes the effect/toxicity evaluation based on in vivo process, providing new ideas and method for screening anti-osteoporosis Chinese medicine.

Quality evaluation of Epimedium sagittatum in different populations based on icariin contents / 中草药

Objective: In order to evaluate the quality of Epimedium sagittatum and to provide the references for screening germplasm as well as for the utilization of resources. The concentration variations of main flavonoids, epimedins A-C, and icariin among 15 representative populations of E. sagittatum and one population of E. sagittatum var. glabratum were assessed by HPLC. Methods: The chromatographic separation was performed on a Zorbax SB-C8 column (250 mm × 4.6 mm, 5 μm). The mobile phase was a mixture of acetonitrile-1.44% acetic acid aqueous solution in gradient elution. The flow rate was 1.0 mL/min, the detection wavelength was 272 nm, and the column temperature was 25 ℃. Results: Remarkable variations within and among the populations were observed in epimedin A (1.00 - 16.64 mg/g), epimedin B (1.00 - 17.21 mg/g), epimedin C (1.00 - 76.21 mg/g), and icariin (1.00 - 46.19 mg/g). As far as the content of icariin concerned, only five populations (HBHF, HBLT, HNLS, HNZJ, and JXWN) had acceptable quality recorded in Chinese Pharmacopoeia, four populations were lower than the standard, and seven populations were even lower than the detectability. Conclusion: The present study suggests that E. sagittatum should not be recorded in Chiese Pharmacopoeia indiscriminately, but the populations of HNZJ, HBLT, HBHF, and HNLS are outstanding and excellent, which were the best candidates for germplasm selection. Meanwhile, their habitat can also provide the reference for cultivation.

Taxonomic research on Epimedium sagittatum species complex and discussion / 中草药

Epimedium sagittatum and its related species are the most controversially defined in Epimedium taxonomy, and are referred as E. sagittatum species complex. So far 11 taxa are included in the complex, but the taxonomical status of most species/varieties is probably open to question. New species based on a single of solely on vegetative characters are of frequent occurrence. Species based on plasticity of phenotype variation should not be regarded as "good species". Based on the investigation in the herbaria, literature, and fields, we found that the morphological variations between some taxa were continuous. Thus E. sagittatum var. guizhouense and E. pudingense were treated as a synonym of E. sagittatum and E. sagittatum var. glabratum, respectively. Six taxa, namely E. coactum, E. coactum var. longtouhum, E. myrianthum var. jianheense, E. multiflorum, E. lobophyllum, and E. jingzhouense were integrated into E. myrianthum. Therefore, the taxonomic problems of E. sagittatum species complex now are back to the relationship between E. myrianthum and E. sagittatum. In morphology, E. myrianthum closely resembles E. sagittatum with only obvious difference in the form of inflorescence, and might be easily confused. Furthermore, in terms of several key taxonomic characters, there was overlapping question between the two taxa, and further research on taxonomy based on more extensive field investigation were needed. A correct species concept stems from a correct and thorough understanding of the nature of variation pattern of plants and its taxonomical value. In general, lack of scientific field investigation and morphological observation, inadequate study on variation range, variation pattern, taxonomical value, and narrow species concept are the reasons for the taxonomical difficulty to deal with.

Effects of total flavonoids of Epimedium sagittatum on the mRNA expression of the estrogen receptor α and β in hypothalamus and hippocampus in ovariectomized rats / 中南大学学报(医学版)

Objective To explore the effect of total flavonoids of Epimedium sagittatum (TFE) on the mRNA expressions of the estrogen receptor alpha (ERα) and estrogen receptor beta(ERβ) in hippocampus and hypothalamus in ovariectomized (OVX) Sprague-Dawley(SD) rats, and the mechanism of TFE against postmenopausal osteoporosis. Methods Forty-eight female SD rats,aged 10-11 months old, were randomly divided into 4 groups: a sham group, an ovariectomy group (rats were bilaterally ovariectomized), a TFE group, and a 17β-estradiol group (rats were fed with TFE and 17β-estradiol for 4 months, respectively). The RT-PCR was performed to determine the mRNA expression of ERα and ERβ in hypothalamus and hippocampus. Results Serum estradiol level, bone mineral density (BMD) of vertebra,wet weight of uterus, and the mRNA expressions of ERα and ERβ in hypothalamus and hippocampus were markedly decreased in OVX rats, all of which were reversed by 17β-estradiol treatment except the mRNA expression of ERβ. Similar results were achieved by TFE treatment except the wet weight of uterus. Conclusion TFE can improve BMD of vertebra in the OVX rats without side effects on the uterus. The mechanism may be related to increasing the mRNA expressions of ERα and ERβ in hypothalamus and hippocampus.

Studies on Extraction of Icariin in Herba Epimedii / 中药新药与临床药理

Objective To optimize the extraction process for icariin from Herba Epimedii. Methods The content of icariin in the extracts obtained by different extraction methods was determined with HPLC. The effect of solvents, boiling time, extracting time and temperature, and the size of the medicinal powder was observed. Results Different extraction methods had great influence on the extraction rate of icariin. Conclusion This research can provide evidence for the extraction of active component from Herba Epimedii in industrial production.

Effect of total flavonoids of Epimedium sagittatum and its metabolites on osteoblast proliferation and function expression / 中国药理学通报

Aim To investigate the effect of total flavonoids of Epimedium sagittatum(TFE)and its metabolites on the proliferation and function expression of newborn rats calvarial osteoblasts(ROB).Methods TFE was supplemented into the culture medium of ROB at 0.2,2,20,100 and 200 mg?L-1 respectively.The serum of rats administered TFES(SRAT) was also added into the medium in a parallel treatment at 2%,4%,8% and 16% respectively.Their effects on cell proliferation and function expression were studied by MTT and the analysis of osteogenic differentiation marks.Results TFE had no effect on cell proliferation and function expression at any concentration.However,2% and 4% SRAT stimulated cell proliferation and,4% SRAT promoted the maturation and function of osteoblast by improving the alkaline phosphatase(ALP) activity,bone gla protein(BGP) secretion,calcium deposition and the number of mineralized nodular structures.Conclusions The effective substance of herba epimedii treating on osteoporosis is not TFE itself but certain of metabolites derived from TFE and the products induced by these metabolites in serum.The metabolites in serum of TFE may enhance the cell function expression and proliferation.