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1.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 254-264, 2023.
Article in Chinese | WPRIM | ID: wpr-996832

ABSTRACT

Paridis Rhizoma, a traditional valuable Chinese herbal medicine, has the functions of clearing heat and removing toxin, relieving edema and pain, cooling liver and calming convulsion, which can be used to treat various diseases such as mumps, abscess, burn, bleeding, and tumor. It has been used in folk medicine for a long time and is the main raw material of various Chinese patent medicines such as Gongxuening Capsules and Yunnan Baiyao. Polyphyllin Ⅰ, an isospirostanol saponin and one of the main active components in Paridis Rhizoma, is distributed in the rhizome, pericarp, and leaves of Paris polyphylla. With high polarity, polyphyllin Ⅰ is mainly extracted by n-butanol extraction and macroporous adsorption resin chromatography, separated by silica gel column chromatography and preparative high performance liquid chromatography, and purified with the combination of methods. With anti-tumor, anti-inflammatory, antibacterial, and anti-virus effects, it is generally employed to treat liver cancer, lung cancer, gastric cancer and other cancers as well as arthritis, influenza, sore toxin, and bacterial infection. However, polyphyllin Ⅰ may cause stomach irritation, hemolysis, liver damage, kidney damage, heart damage, and other adverse reactions. Pharmacokinetic studies show that it has problems such as low bioavailability and poor intestinal absorption and permeability, which affect the clinical application of polyphyllin Ⅰ. This paper summarizes the research on the plant sources, extraction and separation methods, pharmacological effects, adverse reactions, and pharmacokinetics of polyphyllin Ⅰ in recent years, which is expected to provide a reference for the rational clinical application and other in-depth research work of polyphyllin Ⅰ.

2.
Journal of Pharmaceutical Analysis ; (6): 39-54, 2023.
Article in Chinese | WPRIM | ID: wpr-991123

ABSTRACT

Polyphyllin Ⅰ(PPⅠ)and polyphyllin Ⅱ(PⅡ)are the main active substances in the Paris polyphylla.However,liver toxicity of these compounds has impeded their clinical application and the potential hepatotoxicity mechanisms remain to be elucidated.In this work,we found that PPⅠ and PⅡ exposure could induce significant hepatotoxicity in human liver cell line L-02 and zebrafish in a dose-dependent manner.The results of the proteomic analysis in L-02 cells and transcriptome in zebrafish indicated that the hepa-totoxicity of PPⅡ and PⅡwas associated with the cholesterol biosynthetic pathway disorders,which were alleviated by the cholesterol biosynthesis inhibitor lovastatin.Additionally,3-hydroxy-3-methy-lglutaryl CoA reductase(HMGCR)and squalene epoxidase(SQLE),the two rate-limiting enzymes in the choles-terol synthesis,selected as the potential targets,were confirmed by the molecular docking,the over-expression,and knockdown of HMGCR or SQLE with siRNA.Finally,the pull-down and surface plasmon resonance technology revealed that PPⅠ could directly bind with SQLE but not with HMGCR.Collectively,these data demonstrated that PPⅠ-induced hepatotoxicity resulted from the direct binding with SQLE protein and impaired the sterol-regulatory element binding protein 2/HMGCR/SQLE/lanosterol synthase pathways,thus disturbing the cholesterol biosynthesis pathway.The findings of this research can contribute to a better understanding of the key role of SQLE as a potential target in drug-induced hepatotoxicity and provide a therapeutic strategy for the prevention of drug toxic effects with similar structures in the future.

3.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 257-265, 2023.
Article in Chinese | WPRIM | ID: wpr-964967

ABSTRACT

As a rare Chinese medicinal material, Paridis Rhizoma is mainly distributed in Yunnan, Guangxi, and Guizhou in southwestern China, with the effect of clearing heat and detoxifying, alleviating edema and relieving pain, cooling liver and tranquilizing mind. It is particularly effective for injuries from falls, fractures, contusions and strains, snake bites, cold wind-induced convulsion, and other diseases, which has been used for more than 2 000 years. According to modern research, polyphyllin Ⅱ, one of the main active components of Paridis Rhizoma, belongs to diosgenin in structure. It has the anti-tumor, anti-inflammatory, antiviral, antibacterial, immune-regulating, antioxidant, and multidrug resistance-reversing activities, showing good application prospect. Especially, the anti-tumor effect of polyphyllin Ⅱ has attracted wide attention, and the mechanism is inhibiting proliferation, migration, and invasion of tumor cells, inducing cell cycle arrest, apoptosis, and autophagy, suppressing angiogenesis, and modulating tumor microenvironment. However, the pharmacokinetic results show that polyphyllin Ⅱ has low bioavailability in vivo due to the low solubility, poor absorption, unsatisfactory distribution, and slow metabolism, which limit the clinical application. In recent years, there has been an explosion of research on the adverse reactions of polyphyllin Ⅱ, such as the strong hemolytic activity and obvious cytotoxicity to liver, kidney, myocardium and cardiovascular cells. Thus, papers were retrieved from "CNKI", "VIP", "Wanfang Data", "PubMed", "Web of Science", and "Elsevier SD" with "Paris saponin Ⅱ", "Polyphyllin Ⅱ" as the main keywords, and the pharmacological activities and mechanisms, pharmacokinetics, and adverse reactions were summarized. The findings are expected to serve as a reference for the in-depth research, development, and utilization of polyphyllin Ⅱ.

4.
China Journal of Chinese Materia Medica ; (24): 3774-3785, 2023.
Article in Chinese | WPRIM | ID: wpr-981510

ABSTRACT

In this study, the authors cloned a glycosyltransferase gene PpUGT2 from Paris polyphylla var. yunnanensis with the ORF length of 1 773 bp and encoding 590 amino acids. The phylogenetic tree revealed that PpUGT2 belonged to the UGT80A subfamily and was named as UGT80A49 by the UDP-glycosyltransferase(UGT) Nomenclature Committee. The expression vector pET28a-PpUGT2 was constructed, and enzyme catalytic reaction in vitro was conducted via inducing protein expression and extraction. With UDP-glucose as sugar donor and diosgenin and pennogenin as substrates, the protein was found with the ability to catalyze the C-3 hydroxyl β-glycosylation of diosgenin and pennogenin. To further explore its catalytic characteristic, 15 substrates including steroids and triterpenes were selected and PpUGT2 showed its activity towards the C-17 position of sterol testosterone with UDP-glucose as sugar donor. Homology modelling and molecule docking of PpUGT2 with substrates predicted the key residues interacting with ligands. The re-levant residues of PpUGT2-ligand binding model were scanned to calculate the corresponding mutants, and the optimized mutants were obtained according to the changes in binding affinity of the ligand with protein and the surrounding residues within 5.0 Å of ligands, which had reference value for design of the mutants. This study laid a foundation for further exploring the biosynthetic pathway of polyphyllin as well as the structure of sterol glycosyltransferases.


Subject(s)
Ligands , Glycosyltransferases/genetics , Sterols , Phylogeny , Ascomycota , Liliaceae/chemistry , Melanthiaceae , Diosgenin , Sugars , Glucose , Uridine Diphosphate
5.
China Pharmacy ; (12): 1686-1690, 2023.
Article in Chinese | WPRIM | ID: wpr-978958

ABSTRACT

OBJECTIVE To investigate the effects of polyphyllin Ⅵ(PPⅥ) on the proliferation and apoptosis of glioma cells and potential mechanism. METHODS Using human glioma LN229 cells as objects, MTT assay was used to detect the survival rate after treated with different concentrations of PPⅥ [0 (control group), 1, 2, 4, 8, 16, 32, 64 μmol/L] for different time (24, 48, 72 h). The clone formation experiments were adopted to detect the number of cell clones and clone formation rate after being treated with different concentrations of PPⅥ [0 (control group), 2, 4, 8 μmol/L] for 14 days. The flow cytometry and Western blot assay were used to detect the apoptotic rate of cells, the expressions of apoptosis-related protein [B cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), cleaved caspase-3], and the expressions of related proteins of Fas/Fas ligand (FasL) death receptor pathway and protein kinase B (Akt)/glycogen synthesis kinase-3β (GSK-3β) pathway after being treated with different concentrations of PPⅥ [0(control group), 4, 8 μmol/L] for 24 h. RESULTS Compared with the control group, the survival rate of cells, the number of clones and clone formation rate, the protein expression of Bcl-2, and the phosphorylation levels of Akt and GSK-3β protein were decreased significantly in different concentration groups of PPⅥ (P<0.05 or P<0.01). The apoptotic rate, the protein expressions of Bax, cleaved caspase-3, Fas, FasL and cleaved caspase-8 were increased significantly (P<0.05 or P< 0.01). CONCLUSIONS PPⅥ can inhibit the proliferation and induce the apoptosis of human glioma LN229 cells, which may be related to the activation of the Fas/FasL death receptor pathway and the inhibition of the Akt/GSK-3β pathway.

6.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 126-135, 2023.
Article in Chinese | WPRIM | ID: wpr-984590

ABSTRACT

ObjectiveTo study the inhibitory effect of polyphyllin Ⅰ (PPI) on the growth of colorectal cancer cells and its molecular mechanism. MethodRKO cells were cultured and divided into a blank group and PPI treatment groups with concentrations of 0.6, 0.8, 1.0 μmol·L-1, respectively. HRT18 cells were cultured and divided into a blank group and PPI treatment groups with concentrations of 1.2, 1.4, 1.6 μmol·L-1, respectively. The effects of PPI on the proliferation and morphology of colorectal cancer were detected by cell proliferation toxicity assay, trypan blue exclusion assay, plate clone formation assay, and confocal high-intension cell imaging analysis system. Flow cytometry was used to detect the apoptosis rate of colorectal cancer cells. The pQCXIP-GFP-LC3 plasmid transfection assay was used to detect the formation of autophagosomes in colorectal cancer cells after PPI treatment. Western blot was used to detect the expression of apoptosis-related proteins Caspase-3, Caspase-8, and poly ADP ribose polymerase (PARP), the expression of autophagy related protein LC3Ⅱ, and the expression and phosphorylation of Hippo signaling pathway proteins LATS1 and YAP. In the plvx-Flag-YAP plasmid transfection assay, YAP was overexpressed and treated with PPI, and the proliferation of colorectal cancer cells was detected by cytotoxicity assay. The expression of LC3Ⅱ and PARP in colorectal cancer cells was detected by Western blot. SwissADME predicted pharmacokinetic parameters of PPI. ResultAs compared with the blank group, the survival rate and clone formation ability of colorectal cancer cells in the PPI group were significantly decreased (P<0.01), the cell area of colorectal cancer cells in the PPI group was significantly decreased, and the roundness of colorectal cancer cells was significantly increased (P<0.01). As compared with the blank group, the apoptosis rate of colorectal cancer cells in PPI treatment groupw was significantly increased (P<0.01), the expression of apoptotic proteins Caspase-3 and Caspase-8 protein precursor in PPI treatment groups was decreased, and the cleavage of PARP was increased (P<0.01). As compared with the blank group, the expression level of autophagy-related protein LC3Ⅱ in colorectal cancer cells in PPI treatment groups was significantly increased, and the formation of autophagosomes was promoted (P<0.01). As compared with the blank group, the expression of YAP protein in colorectal cancer cells in PPI treatment groups was significantly decreased, and the expressions of phosphorylated LATS1 and YAP were significantly increased (P<0.01). As compared with the blank group, overexpression of YAP could significantly antagonize the effect of PPI on apoptosis, autophagy activation, and proliferation inhibition of colorectal cancer cells. SwissADME simulation results showed that PPI had good drug like activity. ConclusionPPI can induce apoptosis and autophagy of colorectal cancer cells through targeted activation of Hippo signaling pathway, thereby inhibiting their proliferation.

7.
China Journal of Chinese Materia Medica ; (24): 1650-1657, 2022.
Article in Chinese | WPRIM | ID: wpr-928095

ABSTRACT

The present study investigated the mechanism of polyphyllin A(PPA) in inhibiting gastric cancer(GC) cells. GC cells(SGC7901 and MGC803 cell lines) were treated with PPA at different concentrations. The effect of PPA on the proliferation of GC cells was detected by MTT assay, real-time cell analysis(RTCA) assay, and clone-forming assay, respectively. Reactive oxygen species(ROS) of GC cells was detected by flow cytometry. The change of mitochondrial membrane potential was detected by JC-1 assay. The expression and phosphorylation levels of apoptosis-related proteins(caspase-9, caspase-3, and PARP) and proteins related to the signaling pathway(ETS-1, CIP2 A, and Akt) were detected by Western blot. The binding sites of PPA to ETS-1 were analyzed by molecular docking. The affinity of PPA and ETS-1 was detected by drug affinity responsive target stability(DARTS) assay. PPA had a significant inhibitory effect on the proliferation and colony formation of GC cells at a low concentration. The PPA groups showed increased ROS and decreased mitochondrial membrane potential. PPA down-regulated the precursor expression of caspase-9 and caspase-3 and promoted the cleavage of PARP, suggesting that PPA induced the apoptosis of GC cells through the mitochondrial pathway. PPA significantly reduced expression levels of CIP2 A and the phosphorylation of downstream Akt. Molecular docking showed that PPA bound to the ETS domain of ETS-1, the transcription factor of CIP2 A, and formed hydrogen bonds with Pro319 and Asp317. DARTS assay further confirmed that PPA significantly prevented the hydrolysis of ETS-1 by pronase, which was inductive of the direct binding effect of PPA and ETS-1. PPA inhibits the proliferation and induces the apoptosis of GC cells by directly targeting ETS-1 to down-regulate the ETS-1/CIP2 A/Akt signaling pathway.


Subject(s)
Humans , Apoptosis , Cell Line, Tumor , Cell Proliferation , Molecular Docking Simulation , Stomach Neoplasms/metabolism
8.
China Journal of Chinese Materia Medica ; (24): 721-729, 2022.
Article in Chinese | WPRIM | ID: wpr-927955

ABSTRACT

This study aims to investigate the molecular mechanism of polyphyllin Ⅰ(PPⅠ) inhibiting proliferation of human breast cancer cells. Human breast cancer BT474 and MDA-MB-436 cells were treated with different concentrations of PPⅠ, and then the effect of PPⅠ on cell proliferation was detected by MTT assay, trypan blue dye exclusion assay, real-time cell analysis, and clone forming assay, respectively. The apoptosis was detected by Annexin V-FITC/PI staining and then analyzed by flow cytometry. The change of mitochondrial membrane potential was detected by flow cytometry after fluorescent probe JC-1 staining. Western blot was used to detect protein expression and phosphorylation. Molecular docking was performed to detect the binding between PPⅠ and EGFR. The affinity between PPⅠ and EGFR was determined by drug affinity responsive target stability assay. The results indicated that PPⅠ inhibited the proliferation and colony formation of BT474 and MDA-MB-436 cells in a time-and concentration-dependent manner. The PPⅠ treatment group showed significantly increased apoptosis rate and significantly decreased mitochondrial membrane potential. PPⅠ down-regulated the expression of pro-caspase-3 protein, promoted the cleavage of PARP, and significantly reduced the phosphorylation levels of EGFR, Akt, and ERK. Molecular docking showed that PPⅠ bound to the extracellular domain of EGFR and formed hydrogen bond with Gln366 residue. Drug affinity responsive target stability assay confirmed that PPⅠ significantly prevented pronase from hydrolyzing EGFR, indicating that PPⅠ and EGFR have a direct binding effect. In conclusion, PPⅠ inhibited the proliferation and induced apoptosis of breast cancer cells by targeting EGFR to block its downstream signaling pathway. This study lays a foundation for the further development of PPⅠ-targeted drugs against breast cancer.


Subject(s)
Female , Humans , Apoptosis , Breast Neoplasms/genetics , Cell Line, Tumor , Cell Proliferation , Diosgenin/analogs & derivatives , ErbB Receptors , Molecular Docking Simulation
9.
Chinese Journal of Natural Medicines (English Ed.) ; (6): 255-266, 2021.
Article in English | WPRIM | ID: wpr-881069

ABSTRACT

Polyphyllin I (PPI) purified from Polyphyllarhizomes displays puissant cytotoxicity in many kinds of cancers. Several researches investigated its anti-cancer activity. But novel mechanisms are still worth investigation. This study aimed to explore PPI-induced endoplasmic reticulum (ER) stress as well as the underlying mechanism in non-small cell lung cancer (NSCLC). Cell viability or colony-forming was detected by MTT or crystal violet respectively. Cell cycle, apoptosis, reactive oxygen species (ROS) and mitochondrial membrane potential were assessed by flow cytometry. Gene and protein levels were evaluated by qRT-PCR and immunoblotting respectively. Protein interaction was determined by immunoprecipitation or immunofluorescence assay. Gene overexpression or silencing was carried out by transient transfection with plasmids or small interfering RNAs. The Cancer Genome Atlas (TCGA) database was used for Gene Set Enrichment Analysis (GSEA), survival analysis, gene expression statistics or pathway enrichment assay. PPI inhibited the propagation of NSCLC cells, increased non-viable apoptotic cells, arrested cell cycle at G2/M phase, induced ROS levels but failed to decrease mitochondrial membrane potential. High levels of GRP78 indicates poor prognosis in NSCLC patients. PPI selectively suppressed unfolded protein response (UPR)-induced GRP78 expression, subsequently protected CHOP from GRP78-mediated ubiquitination and degradation. We demonstrated that the natural product PPI, obtained from traditional herbal medicine, deserves for further study as a valuable candidate for lead compound in the chemotherapy of NSCLC.

10.
China Pharmacy ; (12): 325-329, 2020.
Article in Chinese | WPRIM | ID: wpr-817338

ABSTRACT

OBJECTIVE:To establish the method for content dete rmination of polyphyllin Ⅱ,Ⅵ,Ⅶ in Ypsilandra thibetica , and to compare the differences of 3 saponins in different parts of Y. thibetica from different producing areas. METHODS :HPLC method was adopted to determine the contents of polyphyllin Ⅱ,Ⅵ,Ⅶ in whole grass part and underground part of Y. thibetica from 10 producing areas. The determination was performed on Kromasil C 18column with mobile phase consisted of acetonirile-water (gradient elution )at the flow rate of 1.0 mL/min. The column temperature was 35 ℃,and detection wavelength was set at 203 nm;sample size was 10 μL. With the contents of 3 saponins as the index ,20 batches of Y. thibetica were analyzed by cluster analysis and PLS-DA analysis ;the aggregation of samples was analyzed and determined the primary difference components. RESULTS:The linear range of polyphyllin Ⅱ,polyphyllin Ⅵ and polyphyllin Ⅶ were 0.051-2.04,0.007-0.28,0.168-6.72 μg, respectively(r≥0.999 5);the detection limits were 1.92,1.75,1.87 ng,respectively;and the quantitative limits were 6.40,5.87, 6.23 ng,respectively;RSD of precision ,reproducibility and stability tests (24 h)were all lower than 2%(n=6);the average recovery rates were 99.29%,101.38% and 99.64%,with RSDs of 1.17%,2.64%,0.75%(n=6),respectively. The content of polyphyllin Ⅱ was 0.615-1.875 mg/g,that of polyphyllin Ⅵ was 0-0.095 mg/g,and that of polyphyllin Ⅶ was 3.158-12.354 mg/g. Cluster analysis showed that 20 batches of samples were clustered into two groups ,batch S 9-S12 were clustered in to one group,and the other 16 batches of samples were clustered into another group. PLS-DA analysis showed that 20 batches of samples were divided into 3 areas,batch S 1,S2,S8,S14,S16,S20 were included in area Ⅰ;batch S 9-S12 included in area Ⅱ;and the rest of the samples included in area Ⅲ. The quality of Y. thibetica from different habitats was different ,and there was no difference in the saponin composition between the whole grass and the underground part. Weight ranking found that mail:cscdtcm@126.com polyphyllin Ⅶ was the main difference component in Y. thibetica,and the content of polyphyllin Ⅶ in Y. thibetica from Pengzhou city and Dayi county was the highest. CONCLUSIONS :The established method is simple ,convenient and sensitive. It can be used for the content determination of 3 saponins in Y. thibetica . The content of active components is higher and the quality is better in Y. thibetica from Pengzhou city and Dayi county.

11.
Chinese Traditional and Herbal Drugs ; (24): 2501-2508, 2020.
Article in Chinese | WPRIM | ID: wpr-846462

ABSTRACT

Objective: To observe the effect of polyphyllin I (PPI) on osteoblasts injuries induced by tricalcium phosphate (TCP) wear particles in vitro, and explain its regulation mechanisms. Methods: Primary osteoblasts obtained from the calvaria of neonatal SD rat by the series of digestion were identified with ALP staining. The osteoblasts were treated with TCP wear particles (TCP, 0.1 mg/mL) for 48 h to establish an in vitro injuries model of the calvarial osteoblasts. The experiment was randomly divided into control group, model (TCP, 0.1 mg/mL) group, PPI (30 μg/mL) group and PPI (100 μg/mL) group. CCK-8 and chemical colorimetry were used to examine cell viability and lactic dehydrogenase (LDH) content in culture media; Real-time PCR was performed to detect mRNA levels of ALP, Collagen I and RUNX2 in osteoblasts; The flow cytometry was used to examine apoptosis of osteoblasts using Annexin V/PI double staining; When the osteoblasts were treated for 14 d, mineral nodules formation was observed with alizarin S staining; Western blot was applied to examine proteins expression of Bax, Bcl-2, cleaved Caspase-3, Atg5, p62, and microtubule associated protein 1 light chain3 (LC-3). Results: Compared with control group, model group showed that the cell viability of osteoblasts, mRNA levels of ALP, Collagen I and RUNX2, and mineral nodules formation were significantly decreased; LDH content, percentage of apoptosis and proteins expression of Bax, cleaved Caspase-3, Atg5, LC-3 and the ratio of LC-3II/LC-3I were obviously increased in calvarial osteoblasts, whereas proteins expression of Bcl-2 and p62 was remarkably decreased. Compared with model group, PPI groups indicated that cell viability of osteoblasts, mRNA levels of ALP, Collagen I and RUNX2, and mineral nodules formation were dramatically increased; LDH content, percentage of apoptosis, protein expressions of Bax, cleaved Caspase-3, Atg5, and LC-3 and the ratio of LC-3II/LC-3I were obviously decreased, but Bcl-2 and p62 expression were obviously increased. Conclusion: PPI alleviates osteoblasts injuries induced by TCP wear particles via inhibition of autophagy.

12.
Tumor ; (12): 163-171, 2020.
Article in Chinese | WPRIM | ID: wpr-848201

ABSTRACT

Objective: To investigate the molecular mechanism of polyphyllin (PP) inhibiting proliferation and inducing apoptosis of human gl i oma U251 cel l s by di rectl y targeti ng si gnal transducer and activator of transcription 3 (STAT3). Methods: U251 cells were treated with different concentrations of PP, then the effect of PP on the proliferation of glioma U251 cells was detected by MTT assay, trypan blue dyeing and clone forming assay, respectively. Apoptosis of glioma U251 cells was determined by flow cytometry after Annexin-FITC/PI double staining. The change of mitochondrial membrane potential was detected by flow cytometry after fluorescent probe JC-1 staining. SwissTargetPrediction online prediction software was used to detect the targeting proteins of PP. Western blotting was used to detect the expressions of STAT3 signaling pathway-related proteins. Molecular docking simulated the binding and action modes of PP and STAT3. Results: PPinhibited the proliferation of U251 cells in a time and concentration-dependent manner (all P 0.05). The hydrogen atom of PP was capable of forming hydrogen bonds with the oxygen atom of aspartic acid (Asp)-334 on STAT3, thereby enhancing its ability of targeting STAT3. Conclusion: PP can inhibit the proliferation of glioma U251 cells and induce apoptosis by directly targeting STAT3.

13.
Chinese Pharmaceutical Journal ; (24): 875-882, 2020.
Article in Chinese | WPRIM | ID: wpr-857680

ABSTRACT

OBJECTIVE: To study the fingerprints of rhizomes of Paris polyphylla Smith var. polyphylla and Paris polyphylla Smith var. yunnanensis (Franch.) Hand. -Mazz from different origins in Dali and the differences of seven main steroidal saponins. METHODS: The fingerprints of rhizomes of Paris polyphylla var. polyphylla and Paris polyphylla var. yunnanensis from different origins in Dali were established by HPLC. The similarity of fingerprints and seven main steroidal saponins were compared and analyzed. RESULTS: There were 14 common peaks in the fingerprints of rhizomes of Paris polyphylla var. polyphylla from different origins in Dali, and the similarity of the fingerprints of the samples from different habitats except Jinhua Weishan and Longjie Weishan was greater than 0.9. There were 13 common peaks in the fingerprints of rhizomes of Paris polyphylla var. yunnanensis from different origins in Dali, and the similarity of the samples from different origins was greater than 0.92. There were 11 common peaks in the fingerprints of rhizomes of Paris polyphylla var. polyphylla and Paris polyphylla var. yunnanensis from different origins in Dali, and the similarity of the fingerprints was very low, between 0.057 and 0.225. Polyphyllin Ⅰ, polyphyllin Ⅱ, polyphyllin Ⅵ and polyphyllin Ⅶ are detected in the rhizomes of Paris polyphylla var. polyphylla and Paris polyphylla var. yunnanensis from different origins in Dali. The average peak areas of polyphyllin Ⅰ, polyphyllin Ⅱ, polyphyllin Ⅵ and polyphyllin Ⅶ in Paris polyphylla var. yunnanensis from different origins in Dali were higher than those of Paris polyphylla var. polyphylla. Polyphyllin H was detected in the rhizomes of Paris polyphylla var. yunnanensis from Leqiu Nanjian, Jinhua Weishan, Fengyu Eryuan, Dengchuan Eryuan, Hongyan Midu, Xiyi Heqing and the rhizomes of Paris polyphylla var. polyphylla from Wanqiao Dali, Fengyi Dali, Xiyi Heqing, Hongyan Midu and Leqiu Nanjian. Polyphyllin Ⅲ was detected in the rhizomes of Paris polyphylla var. polyphylla from the origins except Longjie Weishan. Polyphyllin Ⅴ was also detected in the rhizomes of Paris polyphylla var. polyphylla in Fengyu Eryuan and Xiyi Heqing. CONCLUSION: Polyphyllin Ⅰ, polyphyllin Ⅱ, polyphyllin Ⅵ and polyphyllin Ⅶ are detected in the rhizomes of Paris polyphylla var. polyphylla and Paris polyphylla var. yunnanensis from different origins in Dali. The similarity of the HPLC fingerprints of the rhizomes of Paris polyphylla var. polyphylla and Paris polyphylla var. yunnanensis is very low, and there are great differences in seven main steroidal saponins. The contents of polyphyllin Ⅰ, polyphyllin Ⅱ, polyphyllin Ⅵ and polyphyllin Ⅴ in Paris polyphylla var. yunnanensis are higher than those in Paris var. polyphylla.

14.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 119-123, 2020.
Article in Chinese | WPRIM | ID: wpr-873356

ABSTRACT

Objective:To observe the effect of polyphyllin Ⅰ on the expressions of forkhead box Q1(FOXQ1)and epithelial-mesenchymal transition (EMT)-related factors, in order to explore the possible mechanism of polyphyllin Ⅰ in inhibiting the metastasis of colon cancer. Method:After the treatment with 1.25,2.50 μmol·L-1 polyphyllin Ⅰ on HCT116 cells, Western blot and Real-time PCR were used to detect the expressions of FOXQ1,E-cadherin,Vimentin protein and mRNA. Result:Compared with the blank group, relative expressions of FOXQ1 protein and mRNA in low-concentration polyphyllin Ⅰ group were decreased, while relative expression of E-cadherin mRNA was increased, the differences were not statistically significant, and relative expressions of Vimentin protein and mRNA in low-concentration polyphyllin Ⅰ group were decreased (P<0.05), and relative expression of E-cadherin protein in low-concentration polyphyllin Ⅰ group was increased (P<0.01). Compared with the blank group, relative expressions of FOXQ1, Vimentin protein and mRNA in high-concentration polyphyllin Ⅰ group were decreased,while relative expressions of E-cadherin protein and mRNA were increased (P<0.05, P<0.01). Compared with low-concentration polyphyllin Ⅰ group, relative expressions of Vimentin protein and mRNA in high-concentration polyphyllin Ⅰ group were decreased, but the difference was not statistically significant, relative expressions of E-cadherin protein and mRNA in high-concentration polyphyllin Ⅰ group were increased, whereas relative expressions of FOXQ1 protein and mRNA were decreased (P<0.05, P<0.01). Conclusion:Mechanism of polyphyllin Ⅰ inhibiting the metastasis of colon cancer may be related to the decrease of FOXQ1 and Vimentin expressions, and the up-regulation of E-cadherin.

15.
China Journal of Chinese Materia Medica ; (24): 1418-1422, 2020.
Article in Chinese | WPRIM | ID: wpr-1008587

ABSTRACT

Polyphyllin D is a steroid saponin monomer in Polyphyllin, with antibacterial, analgesic, sedative, anti-tumor and other pharmacological effects, but is rarely reported in pancreatic cancer. This study detected apoptosis-relevant indicators, in order to explore the effect of polyphyllin D on the proliferation and apoptosis of human pancreatic cancer Panc-1 cells and relevant mechanisms of action. After pancreatic cancer Panc-1 cells were treated with polyphyllin D(0, 1, 2, 3, 4, 5 μg·μL~(-1)) for 24, 48 and 72 hours, CCK-8 method was used to detect the effect of polyphyllin D on the proliferation of pancreatic cancer Panc-1 cells. Flow cytometry was used to detect cell cycle and changes in mitochondrial membrane potential(MMP). The apoptosis was detected by Annexin V-FITC/PI staining, and Western blot was used to detect the protein expressions of cytochrome C(Cyto C), Bax, Bcl-2, cleaved caspase-3 and cleaved caspase-9. The results indicated that compared with the control group, polyphyllin D could inhibit the proliferative activity of Panc-1 cells in a time and concentration-dependent manner. Flow cytometry results showed that polyphyllin D could block the cells in S and G_2/M phase in a concentration manner, the MMP of the cells was significantly reduced, and the apoptosis rate increased with the concentration of polyphyllin D. Western blot results showed that polyphyllin D could concentration-dependently up-regulate the protein expression levels of Bax, Cyto C, cleaved caspase-3 and cleaved caspase-9, and down-regulate the protein expression level of Bcl-2. The above findings suggested that polyphyllin D could effectively inhibit the proliferation of Panc-1 cells, and its mechanism may be related to the blocking of cell growth cycle and the apoptosis induced by mitochondrial pathway.


Subject(s)
Humans , Antineoplastic Agents, Phytogenic/pharmacology , Apoptosis , Caspase 3/metabolism , Caspase 9/metabolism , Cell Line, Tumor , Cell Proliferation , Diosgenin/pharmacology , Pancreatic Neoplasms/pathology , Proto-Oncogene Proteins c-bcl-2/metabolism , Saponins/pharmacology , bcl-2-Associated X Protein/metabolism
16.
International Eye Science ; (12): 2028-2033, 2020.
Article in Chinese | WPRIM | ID: wpr-829699

ABSTRACT

@#AIM: To observe the effects of high glucose-induced environment on Visfatin expression in human retinal pigment epithelial cells and to study the effects of Polyphyllin I on Visfatin expression in high glucose environment. <p>METHODS: Human retinal pigment epithelial cells were cultured in three groups, normal control group, high glucose group and intervention group of high glucose aggravated PolyphyllinⅠ, testing after 12h of intervention culture. Normal control group:5.5mmol/L glucose concentration routine culture; high glucose group:25mmol/L high glucose was added to the medium to establish the model; high glucose aggravated PolyphyllinⅠdrug intervention group: high glucose 25mmol/L, 3μg/L PolyphyllinⅠdrug was added to the medium. Immunofluorescence staining assays to observe expression of the Visfatin and VEGF in human retinal pigment epithelial cells; real-time PCR assays for relative expression of Visfatin and VEGF mRNA in epithelial cells; and western-blot assays for Visfatin and VEGF proteins in epithelial cells. <p>RESULTS: Immunofluorescence detection revealed that Visfatin and VEGF were weakly positive in normal retinal pigment epithelial cells. Visfatin and VEGF were strongly positive in high glucose group. Visfatin and VEGF fluorescence in the drug intervention group was significantly weakened in the higher sugar group. RT-PCR showed that the expression level visfatin mRNA high sugar group was significantly higher than that of normal group and intervention group(<i>t</i>=4.24, 3.89, <i>P</i><0.05). VEGF mRNA expression was significantly higher in high glucose group than in normal group and intervention group(<i>t</i>=3.53, 2.57, <i>P</i><0.05). Western-blot results showed that the protein expression of visfatin and VEGF in high sugar group was significantly higher than that in control group and intervention group(<i>t</i>=3.62, <i>P</i>=0.01; <i>t</i>=3.79, <i>P</i><0.01). <p>CONCLUSION: The high glucose environment can stimulate the increased expression of Visfatin in retinal pigment epithelial cells, Polyphyllin I can inhibit the expression of Visfatin in retinal pigment epithelial cells in high glucose environment, which may provide a new idea for the treatment of diabetic retinopathy.

17.
Chinese Critical Care Medicine ; (12): 746-749, 2019.
Article in Chinese | WPRIM | ID: wpr-754048

ABSTRACT

Objective To investigate the protective effect of PolyphyllinⅠ(PPI) on myocardial ischemia/reperfusion (I/R) injury in rats and its mechanism. Methods The 6-month-old Sprague-Dawley (SD) rats were divided into sham operation group (Sham group), I/R model group, and low, medium, high dose PPI groups according to the random number table method, with 10 in each group. The rat myocardial I/R model was prepared by ligating the left anterior descending branch of the coronary artery by 30 minutes and reperfusion by 120 minutes. Sham group was exposure to open chest without ligation. Low, medium, high dose PPI groups were injected with PPI 75, 150, 300 mg·kg-1·d-1 in front of the film for 4 weeks; dimethyl sulfoxide (DMSO) was gastric infused in the I/R model group. After the end of reperfusion, the myocardial infarction area (IA) was determined by triphenyltetrazole (TTC) and Evans blue (EB) staining;the apoptosis of myocardial cells was detected by TdT-mediated dUTP nick end labeling stain (TUNEL); the expressions of apoptosis related protein (Bax, Bcl-2), and cytoplasmic and nucleus expressions of P65 in nuclear factor-κB (NF-κB) signal pathway were detected by Western Blot. Results Compared with the Sham group, the myocardial IA was significantly increased in the I/R model group, the apoptosis rate of myocardial cells was significantly increased, the expression of Bcl-2 was significantly decreased, and the expression of Bax was significantly increased, and the intranuclear transfer of P65 was significantly increased. Compared with the I/R model group, low, medium and high dose PPI pretreatment could significantly reduce the myocardial IA [(21.6±0.9)%, (14.3±1.6)%, (15.0±0.8)% vs. (29.6±1.4)%], the apoptosis rate of myocardial cells was significantly decreased [(38.6±1.9)%, (24.3±2.6)%, (26.3±2.8)% vs. (56.8±2.4)%], the protein expression of Bcl-2 was significantly increased, while the protein expression of Bax was significantly decreased (Bcl-2/GAPDH: 0.24±0.07, 0.36±0.02, 0.34±0.09 vs. 0.13±0.04; Bax/GAPDH: 0.39±0.10, 0.21±0.08, 0.23±0.06 vs. 0.53±0.12); and P65 nuclear transfer was significantly decreased after middle and high dose PPI pretreatment [nuclear P65/Histone 3: 0.49±0.09, 0.51±0.06 vs. 0.83±0.11; cytoplasmic P65/GAPDH: 0.31±0.03, 0.30±0.05 vs. 0.22±0.07], with statistically significant differences (all P < 0.05). However, there was no significant difference in each index between the medium and high dose PPI groups (all P > 0.05). Conclusion PPI alleviates myocardial I/R injury in rats via NF-κB signal pathway, and the PPI effect of 150 mg·kg-1·d-1 is most especially significant.

18.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 93-101, 2019.
Article in Chinese | WPRIM | ID: wpr-801905

ABSTRACT

Objective:To study the appearance description,TLC examination and content determination was carried out, in order to improve the quality standard of processed slices of Paridis Rhizoma in the 2020 edition of Chinese Pharmacopoeia. Method:Based on the literature review and observation on the samples,the appearance description was described. TLC examination was used for the qualitative analysis. HPLC was used for the determination of polyphyllin Ⅰ,Ⅱ,Ⅵ and Ⅶ in the commercial and processed samples. UPLC was employed for the determination of 10 steroidal saponins,namely pseudoprotodioscin,polyphyllin Ⅶ,17-hydroxygracillin,polyphyllin H,polyphyllin Ⅵ,polyphyllin Ⅱ,dioscin,gracilin,polyphyllin Ⅰ and polyphyllin Ⅴ. Result:For the appearance description,color and luster,texture,odor and taste as well as the diameter of 1.0-4.5 cm were recorded. polyphyllin Ⅵ was not detected in the thin layer chromatograms of most of the tested samples derived from high-quality species but obviously detected in those of Trillium Rhizoma. Five of 13 commercial samples met the requirements that the total amounts of polyphyllin Ⅶ,Ⅵ,Ⅱ,and Ⅰ should be no less than 0.6%according to the current Chinese Pharmacopoeia. Because softening and drying had the obvious influence on the contents of steroidal saponins in the samples,soaking and sun-drying were preferred. Conclusion:Appearance description should be supplemented. Polyphyllin Ⅵ is not considered as one of quality markers for the TLC identification and HPLC determination of Paridis Rhizoma. Polyphyllin H was considered as a new marker for the quality control. It is recommended that the total amounts of polyphyllin Ⅶ,H,Ⅱ,and Ⅰ should be no less than 1.0%.

19.
Chinese Traditional and Herbal Drugs ; (24): 526-534, 2019.
Article in Chinese | WPRIM | ID: wpr-851427

ABSTRACT

Objective Mining Paris resources with medicinal value could underlay the breeding selection and relieve the pressure on wild resources. Methods Twenty-one Paris resources in the mountain area of Sichuan Basin were collected and selected according to their phenotype and rhizome features. Two resources GQ, MH with bigger rhizome and one resource (GK) with polygemmic feature were screened. After preliminary identification, based on Kimura-2-parameter model, molecular phylogenetic trees were constructed based on the different sequence of ITS and psbA-trnH between the screened resources and the homologous sequences from NCBI using the Maximum Like (ML) method. Main active saponin was determinated by HPLC method to predict its potential medicinal value. Results GQ, GK, and MH were special resources of P. polyphylla var. chinensis, P. polyphylla var. yunnanensis, and P. forrestii, respectively, in mountains around Sichuan Basin. The content and proportion of polyphyllin I, II, VI, VII in GQ, GK, MH were different. The total content was GQ > MH > CK > GK. The proportion of polyphyllin I in GQ and MH was 67.61% and 73.25% higher than CK, respectively. While the proportion of polyphyllin VII was most in GK (56.38%). Conclusion This study specified three Sichuan local Paris resources with excellent rhizome features. And they performed well after introduced to Chengdu Plain providing the material basis for the follow-up breeding study of Paris. Three resources have medicinal potential, especially the polyphyllin I and polyphyllin content in MH (P. forrestii) is higher, which can provide a new choice for screening substitution materials of P. polyphylla var. chinensis and P. polyphylla var. yunnanensis.

20.
Chinese Traditional and Herbal Drugs ; (24): 3178-3186, 2019.
Article in Chinese | WPRIM | ID: wpr-851028

ABSTRACT

Objective: HPLC was used to determine the content of the polyphyllins VII, H, VI, II, III, I, and V in the rhizomes of multiple stems Paris polyphylla var. yunnanenesis from different places of origin, establishing HPLC fingerprint and comparative analysis HPLC fingerprint between multiple stems P. polyphylla var. yunnanenesis and P. polyphylla var. yunnanensis. Methods: HPLC was performed on the column of Thermore C18 with mobile phase of acetonitrile-water gradient at the flow rate of 1 mL/min; The detection wave-length was 203 nm, column temperature was 30 ℃, and volume was 10 μL; Content of multiple stems P. polyphylla var. yunnanenesis. was measured by external standard and quantitative analysis of multi-components (QAMS), and one-way ANOVA was used to explore its content difference. The HPLC fingerprint of rhizome from multiple stems P. polyphylla var. yunnanenesis was established and compared with that of rhizome from P. polyphylla var. yunnanensis. Results: The determination results of 10 batches of rhizome of multiple stems P. polyphylla var. yunnanenesIs showed a good linear relationship in the linear range (r > 0.999 7), with an average recovery of 98.34%-99.34% and RSD ≦ 1.00%. There was a significant difference (P < 0.05) or highly significant difference (P < 0.01) about content of polyphyllins VII, H, VI, II, III, I, and V among different places of origin in the rhizomes of multiple stems P. polyphylla var. yunnanenesis, the total content of polyphyllins (I+ II + VI + VII)% ranging from 1.239%-6.236%, which was significantly higher than 0.60% of the quality control standard of Paridis Rhizoma prescribed by Chinese Pharmacopoeia. No significant difference was observed between the results of external standard and quantitative analysis of multi-components methods. The HPLC fingerprint similarity evaluation results showed that there were 12 common fingerprint peaks between multiple stems P. polyphylla var. yunnanenesis and P. polyphylla var. yunnanensis, and with the high similarity of HPLC fingerprints. Conclusion: The method was simple, accurate and reproducible, and can be used for the determination of polyphyllins VII, H, VI, II, III, I, and V in multiple stems of P. polyphylla var. yunnanenesis, total content of polyphyllins (I + II + VI + VII)% in multiple stems of P. polyphylla var. yunnanenesis was relatively higher and HPLC fingerprints of rhizome of multiple stems P. polyphylla var. yunnanenesis and P. polyphylla var. yunnanensis from different places of origin also have high similarity.

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