Shuli Man

Chemical study and medical application of saponins as anti-cancer agents.

Saponins are a group of naturally occurring plant glycosides, characterized by their strong foam-forming properties in aqueous solution. The presence of saponins has been reported in more than 100 families of plants out of which at least 150 kinds of natural saponins have been found to possess significant anti-cancer properties. There are more than 11 distinguished classes of saponins including dammaranes, tirucallanes, lupanes, hopanes, oleananes, taraxasteranes, ursanes, cycloartanes, lanostanes, cucurbitanes and steroids. Due to the great variability of their structures, saponins always display anti-tumorigenic effects through varieties of antitumor pathways. In addition, there are a large amount of saponins that still either remain to be trapped or studied in details by the medicinal chemists. This article reviews many such structures and their related chemistry along with the recent advances in understanding mechanism of action and structure-function relationships of saponins at the molecular and cellular levels. These aglycones have been described and their classification and distribution have been listed in the review. Some special saponins with strong antitumor effects have also been exhibited. Ginsenosides, belonging to dammaranes, have been found beneficial targeted on inhibition of tumor angiogenesis by suppressing its inducer in the endothelial cells of blood vessels, and then on prevention of adhering, invasion, and metastasis of tumor cells. Dioscin, one of the steroidal saponins, and its aglycone diosgenin also have been extensively studied on its antitumor effect by cell cycle arrest and apoptosis. Other important molecules discussed include oleanane saponins such as avicins, platycodons, saikosaponins, and soysaponins along with tubeimosides.

Antitumor and antimetastatic activities of Rhizoma Paridis saponins.

Rhizoma Paridis saponins (RPS) have been found to show strong antitumor activity. However, few studies have yet investigated its role on pulmonary metastasis treated with this herb. To investigate the molecular mechanisms related to metastasis, we studied RPS-treated T739 mice using histopathology, immunohistochemistry and reverse transcription polymerase chain reaction. Apoptosis was measured by TUNEL assay. As a result, RPS inhibited tumor growth by inducing apoptosis and upregulated the expression of TIMP-2 and down-regulated the level of MMP-2 and MMP-9. In conclusion, RPS is a potent anticancer agent that elicits programmed cell death and inhibits metastases in murine lung adenocarcinoma in vivo.

Anticancer Drugs from Traditional Toxic Chinese Medicines

Many anticancer drugs are obtained from natural sources. Nature produces a variety of toxic compounds, which are often used as anticancer drugs. Up to now, there are at least 120 species of poisonous botanicals, animals and minerals, of which more than half have been found to possess significant anticancer properties. In spite of their clinical toxicity, they exhibit pharmacological effects and have been used as important traditional Chinese medicines for the different stages of cancer. The article reviews many structures such as alkaloids of Camptotheca acuminata, Catharanthus roseus and Cephalotaxus fortunei, lignans of Dysosma versipellis and Podophyllum emodi, ketones of Garcinia hanburyi, terpenoids of Mylabris and Ginkgo biloba, diterpenoids of Tripterygium wilfordii, Euphorbia fischeriana, Euphorbia lathyris, Euphorbia kansui, Daphne genkwa, Pseudolarix kaempferi and Brucea javanica, triterpenoids of Melia toosendan, steroids of Periploca sepium, Paris polyphylla and Venenum Bufonis, and arsenic compounds including Arsenicum and Realgar. By comparing their related phytochemistry, toxic effects and the recent advances in understanding the mechanisms of action, this review puts forward some ideals and examples about how to increase antitumour activity and/or reduce the side effects experienced with Chinese medicine. Copyright

Characterization of steroidal saponins in saponin extract from Paris polyphylla by liquid chromatography tandem multi-stage mass spectrometry

Rhizoma Paridis saponins are bioactive steroidal saponins derived from Paris polyphylla. Optimization of the ionization process was performed with electrospray ionization tandem mass spectrometry in both positive and negative-ion modes. Negative-ion ESI was adopted for generation of the precursor deprotonated molecules to achieve the best ionization sensitivity for the analytes. Positive ionization was used to choose the most abundant fragment ion. Furthermore, according to the characteristic fragmentation behavior of known steroidal saponins isolated from this plant (polyphyllin D, formosanin C, gracillin, Paris H, Paris VII, and dioscin), 23 constituents were structurally characterized on the basis of their retention time and ESI analyses, including four pairs of naturally occurring isomers. Five of these 23 constituents were new compounds. The analytical method of LC–MSn in positive and negative-ion modes has been developed for the direct structural elucidation of steroid saponins of this kind in plant extracts.

Qualitative and quantitative determination of major saponins in Paris and Trillium by HPLC-ELSD and HPLC–MS/MS

High-performance liquid chromatographic (HPLC) with evaporative light scattering detection (ELSD) and HPLC with electrospray ionization multistage tandem mass spectrometry (HPLC-ESI-MS(n)) were used to identify and quantify steroid saponins in Paris and Trillium plants. The content of the known saponins such as Paris I, II, III, V, VI, VII, H, gracillin and protodioscin in Paris and Trillium plants was determined simultaneously using the developed HPLC-ELSD method. Furthermore, other 12 steroid saponins were identified by HPLC-ESI(+/-)-MS(n) detection. In the end, a developed analytical procedure was proved to be a reliable and rapid method for the quality control of Paris and Trillium plants. In addition, the alternative resources for Paris yunnanensis used as a traditional Chinese medicine were discovered according to the hierarchical clustering analysis of the saponin fraction of these plants.

Identification of chemical constituents in extracts and rat plasma from Fructus Aurantii by UPLC‐PDA‐Q‐TOF/MS

INTRODUCTION  Fructus Aurantii, as a component of several compound formulae, shows many bioactivities. As is well known, the constituents of traditional Chinese medicines are very complex and multiple constituents are responsible for the therapeutic effect. However, the concrete compounds are difficult to confirm. Therefore, studies on the constituents absorbed into serum and their metabolites are necessary. OBJECTIVE To search for the active constituents in Fructus Aurantii. METHODOLOGY An ultra-performance liquid chromatography-photodiode array detector-quadrupole time-of-flight mass spectrometry (UPLC-PDA-Q-TOF/MS) method was established to analyse the aqueous extract in Fructus Aurantii and the constituents absorbed into blood. Compound identification was made by matching the empirical molecular formula with those of the reported compounds and UV spectra, and further elucidated by lower energy CID mass spectra. RESULTS Twelve flavonoid O-glycosides were detected, and nine compounds were tentatively identified as polymethoxylated flavones. Six parent compounds were identified and four metabolites were observed in rat plasma, two of which were identified as naringenin glucuronide and hesperetin glucuronide, respectively. CONCLUSION The approach developed has proved useful in the study of the active constituents in traditional Chinese medicines.

Formosanin C-inhibited pulmonary metastasis through repression of matrix metalloproteinases on mouse lung adenocarcinoma.

Formosanin C (FC) isolated from Rhizoma Paridis, showed pro-apoptosis and immunoregulation with antitumor activity in cultured cells and animal systems. However, there is no report about its anti-metastatic effect on cancer cells. This research used the wound healing and the migration assay to detect the anti-invasive effect of FC on LA795 cells. Through the gelatin zymography assay and immunofluorescence analysis, FC showed suppression of enzyme activity of MMP-2 and MMP-9, and protein expression of MMP-1, -2, -3, -9 and -14 excreted from LA795 cells. Finally, FC exhibited much more effective inhibition of tumor growth and pulmonary metastasis in T739 mice than cisplatin did. Overall, the strong inhibition of MMP expression by FC might provide a potential therapeutic modality for lung tumors.

Anti-fibrosis and anti-cirrhosis effects of Rhizoma paridis saponins on diethylnitrosamine induced rats.

ETHNOPHARMACOLOGICAL RELEVANCE Paris polyphylla var. yunnanensis as a traditional Chinese medicine has been used in the treatment of liver disease for thousands of years. Rhizoma paridis saponins (RPS), as the main active components of Paris polyphylla, have been used to treat liver injury. Anti-cirrhosis effect of Rhizoma paridis saponins (RPS) has not been known. MATERIALS AND METHODS We analyzed diethylnitrosamine (DEN)-induced metabonomic changes in multiple biological matrices (plasma and urine) of rats by using (1)H-NMR spectroscopy together with clinical biochemistry assessments, oxidative stress test and DNA fragmentation assay. RESULTS Mechanisms of RPS that participated in the inhibition of the fibrotic process included anti-oxidant, anti-apoptosis, and metabolic disturbance such as decreasing lipid oxidation, regulation of TCA cycle, carbohydrate, and amino acid metabolisms in DEN-induced liver tissues. CONCLUSIONS Integrated NMR analysis of serum and urine samples, together with traditional clinical biochemical assays provided a holistic method for elucidating mechanisms of potential anti-fibrotic agent, RPS.

Inhibition of matrix metalloproteinases related to metastasis by diosgenyl and pennogenyl saponins.

ETHNOPHARMACOLOGICAL RELEVANCE Diosgenyl and pennogenyl saponins isolated from Rhizoma Paridis, showed pro-apoptosis and immunoregulation with antitumor activity in cultured cells and animal systems. AIM OF THE STUDY To evaluate their anti-metastatic mechanism on cancer cells and discuss their structure-activity relationship on anti-tumor effect. MATERIALS AND METHODS This research used the wound healing and migration assay to detect their anti-invasive effect on B16 melanoma cells. Through the gelatin zymography assay, immunofluorescence analysis and western blot, saponins exhibited different levels of protein expression inhibition of MMP-1, -2, -3, -9 and -14. RESULTS Through the analysis, diosgenyl and pennogenyl saponins inhibited the metastasis of B16 melanoma cells. Diosgenyl saponins also showed strong suppression of enzyme activity of MMP-2 and -9. Different saponins exhibited different levels of inhibition on MMP expression. CONCLUSIONS 17-α OH increases the sensitivity of diosgenyl saponins to the membrane-bound protease which can stimulate proMMP-2 activation, but it also decreases the anti-metastatic activity of diosgenyl saponin. Furthermore, their combination might provide a potential therapeutic modality for metastasis.

Pharmacological evaluation of sedative–hypnotic activity and gastro-intestinal toxicity of Rhizoma Paridis saponins

ETHNOPHARMACOLOGICAL RELEVANCE Rhizoma Paridis saponins (RPS) have been well studied for antimicrobial, anti-hemorrhagic, and anticancer effects. However, scientific information on RPS regarding the toxic and neuropharmacological effects is limited. In this study, the acute oral toxicity, sedative-hypnotic activity and gastro-intestinal toxicity of RPS were investigated. MATERIALS AND METHODS The acute toxicity was carried out by administering single doses (800-5000 mg/kg) of RPS to adult mice. Rotarod test and sodium pentobarbital-induced hypnosis activity were used to evaluate the neuropharmacological effects on mice. Gastric emptying and intestinal transit were used to investigate the gastric-intestinal system effects. RESULTS A single oral administration of RPS dose-dependently caused adverse effects on the general behavior and mortality rate of mice. LD(50) value of oral acute toxicity was 2182.4 mg/kg, with 95% confidence limit of 1718.4-2807.8 mg/kg. In the test of sleeping mice, RPS acted in synergy with sodium pentobarbital at doses 250 and 500 mg/kg while motor coordination was not influenced within 120 min after treatment with RPS. Regarding the gastric-intestinal toxicity, RPS (100, 250, and 500 mg/kg) significantly inhibited gastric emptying but did not affect the intestinal transit. CONCLUSIONS RPS, which is a hypotoxic anticancer drug, possesses the sedative-hypnotic activity and gastric stimulus side effect.