The aim of this paper was to study the effect of total flavones of Clematis filamentosa Dunn(TFCD) post-conditioning against myocardial ischemia-reperfusion injury (MIRI) and the role of PI3K/Akt-eNOS signaling pathway. Forty male SD rats were divided randomly into five groups: Sham group, model group (I/R), TFCD post-conditioning group (TFCD), TFCD post-condition-ing+LY294002 (a PI3K/Akt signaling pathway inhibitor) group (TFCD+LY), and LY294002 group (LY). At the end of reperfusion, hemodynamic parameters were recorded, morphology changes of myocardial tissue were evaluated by using HE staining, and myocardial infarct size were observed, blood samples were obtained to determine plasma activation of lactate dehydrogenase (LDH), creatine kinase (CK) nitric oxide (NO), endothelial nitric oxide synthase (eNOS), superoxide dismutase (SOD), maleic dialdehyde (MDA) and glutathione peroxidase (GSH-Px). The expressions of Akt, p-Akt, eNOS and p-eNOS proteins were assessed by using Western blot, and eNOS and inducible nitric oxide synthase (iNOS) mRNA was measured by RT-PCR. The results showed that, compared with the model group, TFCD post-conditioning remarkably improved hemodynamics function and myocardial structure, reduced myocardial infarct size and enhanced the contents of NO, eNOS, SOD and GSH-Px, and decreased the contents of LDH, CK and MDA, increased the levels of phosphorylation of Akt and eNOS protein expression, eNOS and iNOS mRNA expression significantly(P＜0.05 or P＜0.01). These effects were inhibited by LY294002, a blocker of PI3K/Akt signaling pathway. The above experiments indicated that TFCD post-conditioning could significantly reduce MIRI in rats, the mechanism of which may be associated with increasing antioxidation, scavenging oxygen free radicals, regulating NO generation and activating PI3K/Akt-eNOS signaling pathway.
Animals ; Clematis ; Flavones ; Male ; Myocardial Reperfusion Injury ; Nitric Oxide Synthase Type III ; Phosphatidylinositol 3-Kinases ; Proto-Oncogene Proteins c-akt ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

Four new triterpene saponins, mandshunosides F-I (1-4), together with five known compounds (5-9), were isolated from the roots and rhizomes of Clematis mandshurica. Their structures were elucidated on the basis of spectroscopic evidences and hydrolysis products. Bisdesmosidic saponin (3-9) showed modest suppression of NO production with the inhibition ratios in the range of 51.3%- 64.6% at 50 μmol·L, whereas monodesmosidic saponins with a free carboxyl group at C-28 (1 and 2) showed potent inhibitory activities with IC values being 12.7 and 8.3 μmol·L, respectively.
Animals ; Clematis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Macrophages ; drug effects ; metabolism ; Mice ; Molecular Structure ; Nitric Oxide ; metabolism ; RAW 264.7 Cells ; Rhizome ; chemistry ; Saponins ; chemistry ; isolation & purification ; pharmacology ; Triterpenes ; chemistry ; isolation & purification ; pharmacology

OBJECTIVE: To investigate the anti-proliferative effects of saponins prepared from Plena Clematis (PC) cultured in Fujian Province, China on 4 human tumor cell lines and its possible anti-tumor mechanism. METHODS: The growth inhibition assays of saponins on human esophageal squamous carcinoma cell line (EC9706), human hepatoma cell line (HepG-2), human oral cancer cell line (KB) and human gastric cancer cell line (BGC-823) were evaluated in vitro by thiazolyl blue (MTT) method. The inhibitory effects on EC9706 treated with different concentrations of saponins (15.62, 31.25, 62.50, 125, 250 and 500 μg/mL) were performed in vitro by MTT method. The morphology and nuclear staining with acridine orange/ethidium bromide of EC9706 cells treated with saponins were illustrated under an inverted phase fluorescence microscope. The apoptotic effects of saponins were further evaluated by annexin-V/propidium iodide dual staining experiment to examine the occurrence of phosphatidylserine externalization onto the cell surface by a flflow cytometer. RESULTS: MTT assay showed that the saponins could inhibit the proliferation of 4 tumor cell lines. Among them, the maximum inhibition rate of 73.1% was detected in EC9706 cells at the saponins concentration of 250 μg/mL for 24 h. Further investigation indicated that the saponins induced EC9706 cells apoposis. The EC9706 cells presented apoptotic characteristics when treated with saponins, including that the morphologies of EC9706 cells were appeared round-shaped with higher refraction, and the cell nuclear stained orange with EB after 250 μg/mL saponins exposure. The flow cytometry analysis results showed that the induction of cell cycle arrest in apoptotic system may participate in the anti-proliferative activity of saponins on EC9706 cells. CONCLUSION The saponins from PC exhibited significant cytotoxicity against human EC9706, KB, BGC-823, and HepG-2 cells and might be beneficial to development of ethnic pharmaceutical plant for potential anti-tumor drugs.
Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Cycle Checkpoints ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Clematis ; chemistry ; Humans ; Saponins ; pharmacology

A new stereoisomeric monoterpene glycoside and five already-known compounds were isolated from the n-BuOH soluble fraction of Clematis heracleifolia leaves. On the basis of spectral data, the structures of the isolated compounds were identified as protocatechuic acid (1), ferulic acid (2), caffeic acid (3), aesculin (4), (6Z)-9-hydroxylinaloyl glucoside (5), and 9-hydroxylinaloyl glucoside (6) and these were isolated for the first time from this plant. Among these compounds, (6Z)-9-hydroxylinaloyl glucoside (5) is a newly isolated from plant source.
Clematis* ; Esculin ; Plants ; Stereoisomerism*

L9 (3(4)) orthogonal experiment was used to design the extraction technology of compound Clematidis Radix spray. Weight coefficients of active ingredients and dry extract rate were solved by information entropy. Support vector machine (SVM) was established and the model parameters were optimized through the genetic algorithm. Grid search algorithm was used for optimization of extraction technology of Clematidis Radix spray. The optimal extraction technology was to extract Clematidis Radix spray in water with 6 times the weight of herbal medicine for 3 times, with 2 h once. Bias of value between real and predicted by SVM was 1.23%. SVM was compared with traditional intuitive analysis of orthogonal design. It indicates that the new method used to optimize the extraction parameters of compound Clematidis Radix spray is more accurate and reliable.
Clematis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Support Vector Machine ; Technology, Pharmaceutical

Microscopic identification and NIRS methods were applied to identify Clematidis Radix et Rhizoma of two different origins. The results showed that both methods could identify the samples. NIRS could identify the two samples nondestructively, and provides a basis for establishment of a standard herbs radix clematidis NIRS fingerprint in the future.
China ; Clematis ; chemistry ; classification ; Drugs, Chinese Herbal ; analysis ; chemistry ; Rhizome ; chemistry ; Spectroscopy, Near-Infrared ; methods

Quality survey of different species of Clematidis Radix et Rhizoma was made by determining the content of hederagenin and oleanolic acid from Clematidis Radix et Rhizoma. The result showed that only a few samples of Clematis chinensis met the quality standard for Clematidis Radix et Rhizoma in Chinese Pharmacopoeia 2010 Edition.
Clematis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; standards ; Oleanolic Acid ; analogs & derivatives ; chemistry ; Rhizome ; chemistry

OBJECTIVETo observe the effectiveness and safety of Tongyan spray composed of Chinese medicine for post-stroke dysphagia patients.

METHODOne hundred and twenty-two post-stroke dysphagia patients were randomly assigned to the treatment group (61 cases) and the control group (61 cases). Basic treatment was given to both groups, with Tongyan spray additionally used in oropharynx for the treatment group, and the placebo used for the control group. After 28-day treatment, the clinical effect and safety were evaluated according to the standard swallowing assessment (SSA) scale.

RESULTSOne patient dropped out in each group, and 120 patients reached the final analysis of the study. The total effective rate for the treatment group was 71.7% (43/60), higher than 46.7% (28/60) in the control group (P<0.05), and the improvement on SSA scores of the two groups were significantly different after treatment (P<0.05). For grade 1 dysphagia patients (completely depending on nasogastric tube), the effective rate of the treatment group was 40.9% (9/22), and 12.5% (2/16) of the control group, without significant difference (P>0.05), while the improvement of SSA score was significantly different between the two groups after treatment (P<0.05). For grade 2-3 dysphagia patients (oral and nasogastric tube feeding), the total effective rate of the treatment group was 89.5% (34/38), higher than 59.1% (26/44) in the control group (P<0.05), and also the improvement on SSA scores was significantly different between the two groups after treatment (P<0.05).

CONCLUSIONTongyan spray was an effective and safe method for post-stroke dysphagia patients.

Administration, Inhalation ; Aged ; Aged, 80 and over ; Clematis ; chemistry ; Deglutition Disorders ; drug therapy ; etiology ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Ginger ; chemistry ; Humans ; Male ; Middle Aged ; Stroke ; complications ; Treatment Outcome

To study the chemical constituents of the stems of Clematis parviloba, six compounds were isolated from a 95% ethanol extract by using a combination of various chromatographic techniques including column chromatography over silica gel, ODS, Sephadex LH-20, and semi-preparative HPLC. Two new phenolic glycosides, 2-((E)-3-carboxybut-2-en-yl)-4-hydroxy-3-methyl-phenyl-O-beta-D-glucopyranoside (1) and 4'-hydroxy-phenol-beta-D-[6-O-(4"-hydroxy-3", 5"-dimethoxy-benzoate)] glucopyranoside (2) were isolated, together with a known phenolic glycoside, 4'-hydroxy-3'-methoxy-phenol-beta-D-[6-O-(4"-hydroxy-3", 5"-dimethoxy-benzoate)] glucopyranoside (3) as well as three known megastigmane glycosides, linarionoside A (4), linarionoside C (5), and staphylionoside K (6). Their structures were determined on the basis of spectroscopic analysis and chemical evidence. Among them, compounds 1 and 2 were named as clemaparvilosides A (1) and B (2), respectively, and compounds 3-6 were obtained from Clematis genus for the first time.
Clematis ; chemistry ; Glycosides ; chemistry ; isolation & purification ; Molecular Structure ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry

Progresses in the studies on chemical constituents of Clematis L. (belonging to the family Ranunculaceae) were systematiically reviewed in this article. The plants in this genus have a wide spectrum of constituents as follows: triterpenes, flavonoids, lignans, coumarins, alkaloids, volatile oils, steroids, organic acids, macrocyclic compounds and phenols, etc., among which triterpenoid saponins, flavonoids and lignans are the main components. The triterpenoid saponins are mainly oleanolic type and hederagenin type, most of which are bidesmosidic saponins, substituted with oligosaccharide chains at both C-3 and C-28, and some are substituted with acetyl, caffeoyl, isoferuloyl, p-methoxy cinnamyl and 3,4-dimethoxy cinnamyl groups in the oligosaccharide chains. The flavonoids from Clematis species are mainly flavones, flavonols, flavanones, isoflavones, xanthones and their glucosides (sugar moieties are connected to the aglycone through either the oxygen or the carbon atoms), the aglycones of which are mainly apigenin, kaempferol, luteolin and quercetin. The lignans from Clematis are mainly eupomatene lignans, cyclolignans, monoepoxylignans, bisepoxylignans and lignanolides. Clematis spp. are rich in resources, however, studies on their chemical constituents have only been carried out on twenty or so spp. As a result, it is necessary to expand our study on other spp. from this genus for better utilization of medicinal resources.
Clematis ; chemistry ; Drugs, Chinese Herbal ; chemistry