Anti-colorectal cancer mechanism of Astragali Radix-Curcumae Rhizoma-Paridis Rhizoma based on network pharmacology and experimental verification / 中国中药杂志

The present study explored the underlying mechanism of Astragali Radix-Curcumae Rhizoma-Paridis Rhizoma(AR-CR-PR) in the treatment of colorectal cancer(CRC) by network pharmacology and molecular docking and animal tests and verified the core targets based on the orthotopic transplantation model in nude mice. The active components of AR-CR-PR were retrieved from databases such as TCMSP. The targets of drugs and the disease were obtained from PubChem, SwissTargetPrediction, TTD, and DrugBank, and the intersection targets were imported into STRING for the analysis of the protein-protein interaction(PPI). Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) analyses were performed through DAVID. AutoDock Vina was used to perform molecular docking and binding ability prediction between the active components and the core targets. The effects of AR-CR-PR on tumor growth, metastasis, and phosphorylation of core target proteins in tumor tissues based on the orthotopic transplantation model in nude mice. As revealed by network pharmacology, AR-CR-PR contained nine core components, such as quercetin, curcumin, and β-ecdysone, and the key targets included protein kinase B(AKT1), mitogen-activated protein kinase 3(MAPK3), MAPK1, and epithelial growth factor receptor(EGFR), which was indicated that the anti-CRC effect of AR-CR-PR was presumedly achieved by regulating tumor cell proliferation, apoptosis, migration, and angiogenesis through PI3 K-AKT, MAPK and other signaling pathways. The results of molecular docking showed that the nine core components had strong binding abilities to AKT1 and MAPK3. The results in vivo showed that AR-CR-PR could reduce the volume of the orthotopic tumor, inhibit liver metastasis, and decrease the phosphorylation of AKT1 and MAPK3 in the CRC model. The mechanism of AR-CR-PR in the intervention of CRC may be related to the activation of PI3 K-AKT and MAPK signaling pathway. This study provides a scientific basis for the clinical application of AR-CR-PR in the treatment of CRC and ideas for modern research on AR-CR-PR.

Effects and mechanism of Rhizoma Paridis total saponin on invasion and migration abilities of human gastric adenocarcinoma cell MKN-45 induced by LiCl / 中草药

Objective: To investigate the effect and the possible mechanism of Rhizoma Paridis total saponin (RPTS) on human gastric cancer cell line MKN-45 proliferation, migration and invasion in vitro. Methods: MKN-45 cells were cultured in vitro and treated respectively with indicated concentrations of RPTS (2.5, 5.0, 10.0, 20.0, and 40.0 μg/mL) for 24 h, and cell viability of cell proliferation was detected by MTT assay; The invasive and metastatic ability of MKN-45 treated with indicated concentrations of RPTS (2.5, 5.0, 10.0 μg/mL) was detected by Transwell migration assay and wound healing assay; Elisa assay was employed to detect the concentrations of MMP-9 induced by LiCl after RPTS administration (10, 20, and 40 μg/mL) in the cell supernatant; Western blotting and qRT-PCR were respectively performed to investigate the invasion and migration related protein and mRNA level of VEGF, COX-2, and GSK-3β in RPTS-treated MKN-45 after LiCl stimulation for 24 h. Results: Compared with the control group, RPTS (10, 20, and 40 μg/mL) significantly inhibited the proliferation of MKN-45 cells (P < 0.05 and P < 0.001); RPTS (2.5, 5.0, 10.0 μg/mL) suppressed the invasion and migration of MKN-45 cells (P < 0.05 and P < 0.001); Compared with the model group, RPTS significantly downregulated the expression of MMP-9 in the cell supernatant of MKN-45 cells induced by LiCl (P < 0.05 and P < 0.01), and RPTS also decreased the protein and mRNA expression level of VEGF and COX-2, but it significantly upregulated the expression of GSK-3β at the protein and mRNA level (P < 0.05 and P < 0.001). Conclusion: RPTS play a pivotal role in suppressing the invasion and migration of MKN-45 cells in vitro, and its mechanism may be related to the regulating effects of the Wnt/β-catenin pathway in the human gastric adenocarcinoma cell.

Study on the correlation among the species,growth conditions and the quality of Paridis polyphylla / 国际中医中药杂志

Objective To explore the relation among the quality of the Paridis polyphylla and different species and growth conditions, and help to provide scientific foundation for introduction and cultivation of Paridis polyphylla.Methods The samples of Paridis polyphylla were collected by different varieties, different growth years, different harvest seasons and different altitude habitats. The content of saponins in Paridis polyphylla was measured by HPLC method according to the Chinese Pharmacopoeia 2015. The chromatographic column was ACQUITY UPLC? HSS T3 (3.0 mm× 100 mm, 1.8μm), the mobile phase was acetonitrile and water (gradient elution), with flow rate of 0.5 ml/min, detection wavelength was 203 nm and column temperature at 35℃.Results Total saponin content ranged from 1.35% to 3.89% among the varieties studied. The content of each components were listed as:Paris polyphylla Smith(PS) >Paris polyphylla Smith var. yunnanensis(Franch.) Hand.-Mazz. (PY) >Paris polyphylla Smith var. chinensis(Franch.) Hara (PC). From 3 to 10 years old of PC, the longer the growth years made the higher the total saponin content. Furthermore, total saponin content of PC increased gradually with the altitudes rising from 400 to 800 meters. The total saponin content of PY harvesting in spring was much higher than that of other seasons.Conclusions The Results showed the importance for introducing and cultivating of Paridis polyphylla.

Determination of diosgenin in Rhizoma Paridis by HPLC and TLC / 中草药

Object To determine the contents of diosgenin in Rhizoma Paridis by HPLC and TLC. Methods ODS column was used in HPLC with mobile phase: acetonitrile water(90∶10), detection wavelength: 203 nm, column temperature: 35 ℃; silica gel plate was used in TLC with developer: cyclohexane ethyl acetate (4∶1), scan wavelength: 610 nm, reference wavelength: 420 nm. Results The linear ranges of HPLC and TLC were 1 2 - 7 2 ?g (r=0 999 8), 0 2 - 1 0 ?g (r=0 995 7). The average recovery and RSD were 102 2%, 3 39% (n=6) and 100 9%, 2 68% (n=6) respectively. Conclusion Both of two methods can be used for the quality control of diosgenin in Rhizoma Paridis.

Antitumor activity of steroid saponins extracted from Paris polyphylla var.yunnanensis against lung adenocarcinoma cells in vitro and in vivo / 中草药

Objective To study the antitumor activity of steroid saponins,such as Rhizoma Paridis saponins,Rhizoma Paridis saponin Ⅰ,and diosgenin,extracted from Paris polyphylla var.yunnanensis against lung adenocarcinoma cells.Methods Taking mouse lung adenocarcinoma LA795 cell line and its T739-bearing mice as models,the antitumor activity of steroid saponins of P.polyphylla var.yunnanensis against lung adenocarcinoma cells was carried out in vitro and in vivo.The inhibitory activity of Rhizoma Paridis saponins,Rhizoma Paridis saponin Ⅰ,and diosgenin on the LA795 cell line was studied by means of MTT assay and their antitumor effects in vivo were carried on inbred strains of mice(T739) affected by LA795 metastatic lung cancer.Results The MTT assay of in vitro cytotoxicity of Rhizoma Paridis saponins,Rhizoma Paridis saponin Ⅰ,and diosgenin showed strong cytotoxicity against the growth of LA795 cells.IC50 Values were 24.33 ?g/mL,1.85,and 149.75 ?mol/L,respectively.The tumor weight in all treated groups was significantly lower than that in the control group and the inhibition of tumor growth were 41.82%,29.44%,and 33.94% in their high-dose groups.The metastasis of tumor cells was inhibited by different degrees in treated groups.Conclusion Steroid saponins of P.polyphylla var.yunnanensis have tumoricidal activity on lung adenocarcinoma cell line,both in vitro and in vivo.

Determination of Anti tumor Cytotoxic Active Substance Gracillin in Rhizoma Paridis and Yunnan White / 中成药

Objective:To establish a HPLC for determination of Gracillin in Rhizoma Paridis and Yunnan white on the bases of determining anti tumor cytotoxic bioactivities of more than 20 varieties of chinese medical materials and traditional Chinese patent medicines.Methods: The determination was carried out by HPLC on KR 100-5C18 column, with acetoitrile water (43∶5) as a mobile phase. The detection wavelength was 210nm. and the flow rate was 0.8ml/min.Results: The recoveries are 99.8% (RSD in 2.6%, n=6) and 98.8% (RSD is 1.5%, n=6), respechrely. The within day and between day accuracies are 1.4% of RSD(n=6) and 1.8% of RSD (n=4), respectively.Conclusion: The results showed that water, methanol and alcohol extracts of Rhizoma Paridis and Yunnan white had strong inhibition on 6 kinds of neoplasm cells of human body and their component Grarillin also had the inhibition on tumor cells.

Study on IR fingerprint of 13 kinds of Chinese medicinal granules / 中成药

AIM: To identify the Chinese medicinal granule by measuring IR fingerprints. METHODS: 13 kinds of granules were extracted with butanone respectively and then the obtained extracts were measured with the FT IR spectrometer. RESULTS: By IR fingerprint of 13 kinds of granules drugs, different batches of the same crude drug had a stable and repeatable fingerprint. CONCLUSION: By using IR fingerprint, Chinese medicinal granule can be identified and provides a rapid method for drug identification and quality control.