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UHPLC-Q-TOF/MS metabonomics technology was used to clarify the metabolic regulation pathways by which Platycodon total saponins (PTS) exert antitussive and expectorant effects in a mouse cough model, in which coughing is induced by concentrated ammonia, and in a phenol red excretion model. After approval by the Experimental Animal Ethics Committee of Jiangxi University of Chinese Medicine (Approval No. JZLLSC-20190235), the mice were randomly divided into a normal group, a model group, a positive drug group and a PTS group. Endogenous metabolites in mouse serum were identified by UHPLC-Q-TOF/MS. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used for multivariate analysis. Metabolic pathways were analyzed by the Metaboanalyst platform. The results show that PTS can significantly prolong the cough latent period and cough frequency of mice, and significantly increase phenol red excretion. UHPLC-Q-TOF/MS identified 19 metabolites related to cough, and PTS significantly decreased 16 of them; 17 metabolites related to expectoration were identified, and PTS decreased the levels of all. Metabolic pathway analysis showed that linoleic acid metabolism, arachidonic acid metabolism and glycerophospholipid metabolism were the main pathways involved in serum metabolite changes in this mouse cough model. Linoleic acid metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, arachidonic acid metabolism, phenylalanine metabolism and α-linolenic acid metabolism were the main pathways involved in serum metabolite changes in the phenol red excretion model. This study is the first to elucidate the regulation of antitussive and expectorant metabolic pathways and the effect of PTS on these pathways.
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AIM: To study the anti-inflammatory and analgesic effects of total saponin of Dioscorea(TSD). METHODS: The rat acute gouty arthritis (AGA) and the mice air-pouch inflammation models were induced by monosodium urate(MSU), the pain models were adopted including the hot-plate and acetic acid-induced writhing test in mice. Then the content of IL-1β, IL-18 and PGE2 levels were determined by ELISA. RESULTS:TSD significantly inhibited the MSU-induced ankle swelling and air pouch inflammation in mice(P<0.01 or P<0.05). Meanwhile, TSD markedly reduced the levels of IL-1β and IL-18 in synovial fluid exudates and IL-1β and PGE2 in air pouch exudates (P<0.01 or P<0.05).In addition, TSD remarkably suppressed acetic acid-induced writhing response and prolonged pain threshold in hot plate assay(P<0.01 or P<0.05).CONCLUSION: TSD has anti-inflammatory and analgesic effects, which are mainly relevant to the inhibition of the synthesis and release of inflammatory factors IL-1β, IL-18 and PGE2.
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Objective: To investigate the relationship between basic nutrient, the activity of soil enzyme and the quality of rhizome in the rhizosphere soil of wild and transplanted Paris polyphylla var. yunnanensis, and select the primal soil factors affecting the active components of P. polyphylla var. yunnanensis, and provide the reference for the rational fertilization of P. polyphylla var. yunnanensis. Methods: A total of 33 samples of rhizome and soil collected from P. polyphylla var. yunnanensis in Yunnan, Guizhou and Sichuan were used as materials. The basic nutrient, the activity of soil enzyme and the quality of rhizome of the samples were measured. On this basis, the correlation between the quality of rhizome and soil factor was analyzed. The primal soil factors affecting the quality of rhizome of P. polyphylla var. yunnanensis were also selected by using the method of stepwise regression analysis. Results: There were some differences in the contents of the total saponins, the total polysaccharides and the total flavonoids between the samples of wild and transplanted P. polyphylla var. yunnanensis, and there were also regional characteristics in quality between them. In all the rhizosphere soil samples of P. polyphylla var. yunnanensis, the pH of the soil was moderate, the basic nutrient was sufficient and rich, and the activity of soil enzyme was high, which were suitable for the growth and development of P. polyphylla var. yunnanensis. And the soil fertility quality of the wild P. polyphylla var. yunnanensis was better than the transplanted. The content of total saponins in the wild P. polyphylla var. yunnanensis was significantly positively correlated with the activity of polyphenol oxidase, and the content of total polysaccharides was significantly negatively correlated with the pH of the rhizosphere soil and the activity of urease. The content of total saponins in the transplanted P. polyphylla var. yunnanensis was negatively correlated with the content of available potassium in rhizosphere soil, the content of total flavonoids was significantly positively correlated with the activity of sucrase, and the content of total polysaccharides was significantly positively correlated with the activity of alkaline phosphatase. Conclusion Based on the traditional evaluation, the quality of the transplanted P. polyphylla var. yunnanensis samples is equal to the wild samples, they can be used as the same in the market. The quality of P. polyphylla var. yunnanensis is mainly affected by the soil factors. In the process of protecting wild P. polyphylla var. yunnanensis and cultivating P. polyphylla var. yunnanensis, it is need to be careful to increase and decrease the content and the ratio of soil nutrient according to the actual conditions.
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Objective::To investigate the effect of total saponin of Dioscoreae Collettii Rhizoma (TSD) on Toll-like receptor/nuclear factor-κB (TLR/NF-κB) signaling pathway induced by monosodium urate in THP-1 cells, in order to explore the possible mechanism of anti-gout arthritis. Method::Phorbol 12-myristate 13-acetate (PMA)-induced THP-1 cells were differentiated into macrophages, divided into normal group, model group, low, medium and high-concentration TSD groups (1, 3, 10 mg·L-1) and colchicine group (0.2 mg·L-1). Except the normal group, the other groups were stimulated with 400 mg·L-1 monosodium urate to replicate an inflammation model in vitro. Cell viability was measured by methyl thiazolyl tetrazolium (MTT) assay, the levels of inflammatory factors tumor necrosis factor-α(TNF-α ) and interleukin-1β(IL-1β) were detected by enzyme-linked immunosorbent assay (ELISA). The protein levels of Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88) and NF-κB were detected by Western blot. The mRNA levels of TLR4, NF-κB and Pro-IL-1β were measured by real-time fluorescence quantitative PCR (Real-time PCR), and the nuclear shift of NF-κB p65 was detected by immunofluorescence. Result::0~32 mg·L-1 TSD has no effect on cell viability. Compared with the normal group, the secretion levels of inflammatory factors TNF-α and IL-1β in the model group were significantly increased (P<0.01), and the expressions of key proteins (TLR4, MyD88 and NF-κB) and genes (TLR4, NF-κB and Pro-IL-1β) were increased (P<0.01). Compared with the model group, 1-30 mg·L-1 TSD significantly down-regulated the secretion of inflammatory factors TNF-α and IL-1β (P<0.01), the expressions of key proteins (TLR4, MyD88 and NF-κB) and genes (TLR4, NF-κB and Pro-IL-1β) were decreased (P<0.05, P<0.01), and the NF-κB p65 partially trans-located to the cytosol and the superposition in the nucleus were decreased, inhibiting the nuclear translocation of NF-κB p65. Conclusion::TSD may exert an anti-inflammatory effect by down-regulating the expressions of TLR4, NF-κB and Pro-IL-1β mRNA and reducing the secretion of inflammatory factors TNF-α and IL-1β.
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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.
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Objective: To research the effects of different gradient levels of N, P, and K of rhizome of Paris polyphylla var. chinensis yield and quality, determine the optimal fertilization scheme and provide a scientific basis for rational fertilization. Methods: The three-factor five-level quadratic general rotation combination design was adopted for field experiment, correlation measurement and data analysis were carried out, and the effects of combined application of N, P, and K on weight gain rate, total saponin content, ash, moisture and extracts were investigated. Results: The established regression equations reached significant levels. When applying 41.66-52.60 kg/hm2 of N, 181.38-244.38 kg/hm2 of P, 24.16-35.20 kg/hm2 of K, the rhizome weight gain rate of P. polyphylla var. chinensis was greater than 81.72%; When applying 45.48-53.83 kg/hm2 of N, 179.98-236.83 kg/hm2 of P, 29.80-39.95 kg/hm2 of K, the total saponin content in the rhizome of P. polyphylla var. chinensis was greater than 11.09%; When applying 28.01-37.79 kg/hm2 of N, 127.18-209.18 kg/hm2 of P, 25.27-34.09 kg/hm2 of K, the extract content of the rhizome of P. polyphylla var. chinensis was more than 14.31%. Conclusion: The optimum fertilization amount on the rhizome of P. polyphylla var. chinensis was as follow: 37.79-45.48 kg/hm2 of N, 181.38-209.18 kg/hm2 of P, and 29.80-34.09 kg/hm2 of K.
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Objective To study the apoptosis of human colorectal cancer HT-29 cells induced by Aralia elata Seem leaf total saponin (ETS) and its effects on the expression of relevant proteins. Methods MTT assay was used to detect the proliferation of human colorectal cancer HT-29 cells cultivated with different concentrations (6.25, 12.5, 25, 50, 100, 200 mg/kg) of ETS. Hoechst33258 staining and laser confocal imaging were used to detect the apoptotic cells. Morphological changes were observed. The expressions of Bcl-2 and Bax were detected by immuno-histochemistry. Results ETS could induce apoptosis of HT-29 cells and apoptosis was in a dose-dependent manner in a certain range. ETS could decrease the expression of Bcl-2 and increase the expression of Bax in HT-29 cells (P<0.05, P<0.01), showing a significant dose-effect relationship. Conclusion ETS can induce the apoptosis of HT-29 cells, and the mechanism may be related to reducing the expression of Bcl-2 and increasing the expression of Bax.
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Aim Toinvestigatetheeffectsoftotalsap-onin of Dioscorea (TSD)on rats with monosodium u-rate crystal-induced acute gouty arthritis (AGA)and mechanisms.Methods Totally72Wistarratswere randomly devided into six groups,Each group was giv-en corresponding drug before,then rat acute gouty ar-thritis model was made by injection of monosodium u-rate in the ankle joint cavity.The gait,articular swell-ing degree and physiological changes of rats were ob-served.The concentration of TNF-α,IL-1β,IL-18 in serum were detected by ELISA.The levels of pro-IL-1β, NALP3, ASC, pro-caspase-1, and cleaved caspase-1 were detected by Western blot.Results AllTSDgroupsandcolchicinesignificantlychangedthe gait of rats and TSD high and middle groups signifi-cantly reduced joint swelling and diminished the patho-logical changes.The levels of TNF-α,IL-1β,IL-18 in serum were significantly decreased,and the levels of pro-IL-1β,NALP3,ASC,pro-caspase-1 and cleaved caspase-1 were apparently reduced in TSD high and middlegroups.Conclusion TSDpossessesanti-goutfunction and the mechanism may be related to sup-pressing the NALP3 inflammasome activation and in-hibiting the cytokine production.
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OBJECTIVE: To optimize the formulation of self-microemulsion containing total saponins from Lysimachia capillipes by central composite design-response surface methodology. METHODS: Based on the study of solubility, compatibility test and ternary phase diagram, central composite design-response surface methodology was adopted to optimize the best prescription with the emulsifying time, particle size, and Zeta pontential as indexes, physicochemical properties of this self-microemulsion were also determined. RESULTS: Optimum formulation was 12.93% of ethyl oleate, 57.25% of Kolliphor RH40, 29.82% of Transcutol. The a verage particle size, polydispersity index, Zeta pontential and drug loading of self-microemulsion containing total saponins from Lysimachia capillipes were 23.0 nm, 0.160, -20.28 mV and 10.52 mg·g-1.CONCLUSION: With good predictability, this methods can be used for the formulation optimization of self-microemulsion loaded total saponins from Lysimachia capillipes.
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Objective To observe the effects of saponins from Panax japonicus on neuronal apoptosis of natural aging rats and its mechanisms based on NLRP1 and NLRP3 inflammasome pathway. Methods Male SD rats in a SPF grade were randomly divided into five groups: control group (9-month-old rats), model group (24-month-old rats), and SPJ treatment group (10, 30, and 60 mg/kg). From the beginning of 18 months, animals were treated with SPJ (or normal saline) by ig until 24 months, and stopped 2 d each week for six months of continuous administration. The neural apoptosis situation of cortex and hippocampus in aging rats were observed by TUNEL method. The protein expression of IL-1β, ASC, NLRP3, NLRP1, Caspase-1, and IL-18 of the cerebral cortex and hippocampal were detected by Western blotting. Results TUNEL results showed that there were a very small number of apoptotic cells in the cortex and hippocampus in control group. Compared with control group, the model group significantly increased the number of apoptotic cells. Compared with model group, the number of apoptotic cells was significantly decreased in rat cortex and hippocampus (CA1, CA3, and DG) after treated with SPJ (10, 30, and 60 mg/kg). Western blotting results showed a significant age-related increase in the expression of IL-1β, ASC, NLRP3, NLRP1, Caspase-1, and IL-18, while SPJ concentration-dependently decreased the levels of IL-1β, ASC, NLRP3, NLRP1, Caspase-1, and IL-18 after six-month treatment. Conclusion In conclusion, saponins from P. japonicus has protective effects on the brain (cortex and hippocampus) of aging rats. The mechanism is likely to be that saponins from P. japonicus can reduce nerve inflammation by regulating NLRP1 and NLRP3 inflammasome pathway.
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Objective To study the clinical treatment effect of intraoperative perfusion of Panax notoginseng saponins on the postoperative mammary gland hematoma and skin ecchymosis. Methods A total of 100 patients who underwent breast minimally invasive surgery from August 2015 to May 2016 in Wenling First People's Hospital were randomly divided into control group and experimental group 50 cases in each group.The control group underwent intraoperative perfusion of Panax notoginseng saponin,on the basis of this, the experimental group were given psychological intervention, pay attention to the psychological state of the patients, strengthen the communication and exchanges with the patients, increased the patient's treatment confidence and treatment compliance. The clinical indicators of the experimental group and the control group were compared. Results After the corresponding treatment, the incidence of postoperative hematoma in the experimental group (4.0%) was significantly lower than in the control group (12.0%), the difference was statistically significant (P<0.05). The probability of skin staining in control group (26.0%) was significantly higher than that in the experimental group (8.0%), the difference was statistically significant (P<0.05). Conclusion Breast minimally invasive biopsy perioperative psychological intervention, intraoperative perfusion of panax notoginseng saponins can reduce the incidence of postoperative mammary gland hematoma, skin ecchymosis.
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Objective To study the clinical treatment effect of intraoperative perfusion of Panax notoginseng saponins on the postoperative mammary gland hematoma and skin ecchymosis. Methods A total of 100 patients who underwent breast minimally invasive surgery from August 2015 to May 2016 in Wenling First People's Hospital were randomly divided into control group and experimental group 50 cases in each group.The control group underwent intraoperative perfusion of Panax notoginseng saponin,on the basis of this, the experimental group were given psychological intervention, pay attention to the psychological state of the patients, strengthen the communication and exchanges with the patients, increased the patient's treatment confidence and treatment compliance. The clinical indicators of the experimental group and the control group were compared. Results After the corresponding treatment, the incidence of postoperative hematoma in the experimental group (4.0%) was significantly lower than in the control group (12.0%), the difference was statistically significant (P<0.05). The probability of skin staining in control group (26.0%) was significantly higher than that in the experimental group (8.0%), the difference was statistically significant (P<0.05). Conclusion Breast minimally invasive biopsy perioperative psychological intervention, intraoperative perfusion of panax notoginseng saponins can reduce the incidence of postoperative mammary gland hematoma, skin ecchymosis.
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ObjectiveTo study the effects of 9 different drying methods on total saponin content and antioxidant capacity ofParis polyphylla var.yunnanensis.Methods UV-VIS spectrophotometry was used for measuring the total saponin content ofP. polyphylla var.yunnanensis, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzthiozoline-6)-sulphonic acid (ABTS+) radical scavenging methods were used for evaluating the antioxidant capacity ofP. polyphylla var. yunnanensisextracts.ResultsThe total saponin contents ofP. polyphylla var.yunnanensis dried by different methods existed significant difference. The sample dried at 35℃ showed highest total saponin content (17.557 mg/g). The sample dried in the shade naturally showed middle total saponin content (13.740 mg/g). Based on the results of antioxidant capacity detected by two methods, the samples ofP. polyphylla var. yunnanensis dried by 9 different methods all showed good antioxidant capacity and showed a certain concentration dependence.Conclusion The samples dried at 35℃ have the highest antioxidant capacity. Therefore, drying at 35℃ is a better drying method to obtain high quality and high activity ofP. polyphylla var.yunnanensis.
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Objective To investigate the anti-tumor effects of total saponins,extracted from stems and leaves ofParispolyphyllavar.yunnanensis. Methods Taking mouse stomach carcinoma MFC cell line,human mammary cancer cell line(MCF-7)and human cervical carcinoma cell line(Hela),and their tumor-bearing mice as models,the antitumor activity was carried out in vitro and in vivo. CCK-8 assay was used to determine the inhibitory effect in vitro. The tumor-bearing mice model was induced by tumor cell vaccination in normal mouse forelimb left axillary subcutaneous and these mice were randomized into NS group,cytoxan group(30 mg/kg),saponins high-dose,medium-dose and low-dose groups(60,30 and 15 mg/kg). They were given intraperitoneal injection once a day for consecutive 15 days. The tumor inhibition rate and survival of the tumor-bearing mice were measured. Results Saponins,extracted from both aboveground and underground ofP.polyphyllavar.yunnanensis could significantly inhibit the growth of MFC,MCF-7 and Hela cells in time- and dose-dependent manners. The tumor weight in each drug treated group was significantly lower than that in the control group. Conclusion Saponins,extracted from both aboveground and underground ofP.polyphyllavar.yunnanensis have antitumor activity.
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OBJECTIVE:To optimize extraction technology of saponins from Paris polyphylla. METHODS:Using paris sapo-nin Ⅰ,paris saponin Ⅱand Paridis total saponin as dependent variables,using ethanol volume fraction,extraction time and sol-vent amount as independent variables,through multiple linear regression and binomial fitting,the extraction technology was opti-mized with response surface method and predicted. RESULTS:The optimized extraction technology of saponins from P. polyphylla was as follows as 10-fold of 80% ethanol,2 times reflux extraction,100 min each time. Under the extraction technology,the ex-traction rates of paris saponinⅠwere 85.4%,82.7% and 87.1%;those of paris saponinⅡwere 85.9%,81.3% and 83.6%;and those of Paridis total saponin were 89.5%,92.1% and 90.3%(all RSD<2.0%). Measured value was 0.964 9,predicted value was 0.986 0 and deviation rate was 2.14%. CONCLUSIONS:The central composite design-response surface method is simple and reliable for the optimization of extraction technology of saponins from P. polyphylla.
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This study was aimed to establish the determination method of total saponins content in Jiu-Bi-Y ing Y i-Xin (JBYYX) tablet. The content of total saponins in JBYYX tablet was determined by ultraviolet spectrophotometry. The results showed that in the range of 2.16í10-5-6.47í10-5 mg·mL-1, the total saponins had good linear relation-ship. The average recycle rate was 98.81%. RSD was 2.06%(n=6). It was concluded that the method was accurate, reliable and repeatable, which was suitable for determining the content of total saponins in JBYYX tablet.
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OBJECTIVE: To establish a rapid and nondestructive method to determine coating thickness of the total saponin of Radix Bupleuri enteric coated tablets by near infrared spectroscopy (NIRS) analytical technique. METHODS: Spectral pretreating methods, wavenumber selection and factors of NIRS were discussed in detail. Different modeling methods were compared and the methodology of model was investigated. RESULTS: Partial least squares regression (PLSR) was used for building the quantitative calibration model. Correlation coefficients(r), root-mean-squares error of cross-validation (RMSECV), and root mean square error of prediction (RMSEP) obtained by PLSR model were 0.9915, 3.6 and 3.24 μm respectively. CONCLUSION: Predicted results show that the established method is rapid, nondestructive, and reliable, which can be applied to the online monitor of Chinese medicine tablets coating process.
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CONCLUSION: Compared with traditional tannin removing process, chitosan flocculation purification process can effectively retain the active ingredients in the solution and has good process stability, which is suitable for industrial production.
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Sanqi is a popular traditional Chinese medicine and commonly used for promoting blood circulation and removing blood stasis. Notoginsenoside R1, ginsenoside Rg1, Re, Rb1 and Rd are the major active constituents of Sanqi. The purpose of the study was to investigate the pharmacokinetic behavior of the five active constituents from total saponin from Sanqi when it was used in the blood stasis animals or in combination with Gegen. The concentrations of the five active constituents in rat plasma were determined by an ultra-HPLC-ESI-MS/MS method. The main pharmacokinetic parameters were calculated and statistically analyzed using the unpaired student's t-test. It was found that the pharmacokinetic parameters of notoginsenoside R1, ginsenoside Rg1 and Rb1 represented a statistically significant difference (Po0.05) between the normal rats and the blood stasis rats after administration of total saponin from Sanqi (TSFS). And there were statistically significant differences (Po0.05) in the pharmacokinetic parameters of all the five constituents between administration of TSFS alone and combined with total flavonoid from Gegen (TFFG) in blood stasis rats. It suggested that the pharmacokinetic behavior of the active constituents from TSFS could be changed when it was used in blood stasis animals or in combination with TFFG.
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OBJECTIVE: To study the effects of total saponin of Rhizoma Dioscreae Nipponicae (TSRDN) on the expression of vascular endothelial growth factor (VEGF), angiopoietin-2 (Ang-2) and receptor Tie-2 in synovial tissue of collagen-induced arthritis (CIA) rats, and to investigate the antiangiogenesis mechanism of TSRDN in treating rheumatoid arthritis. METHODS: After the CIA rat model was successfully established, the rats were randomly divided into 5 groups: normal control group, CIA model group, TSRDN group, tripterygium group, and diosgenin group. Real-time PCR was used to detect VEGF mRNA expression in synovial tissue of CIA rats, and immunohistochemical staining was used to observe angiopoietin-2 and receptor Tie-2. RESULTS: VEGF mRNA, Ang-2 and Tie-2 expressions in synovial tissue of CIA rats were obviously higher than normal control group (P < 0.01). After treatment with TSRDN, tripterygium, and diosgenin, the expressions of VEGF mRNA and Ang-2 were obviously lower than those in CIA model group (P < 0.01). But Tie-2 expression showed decreasing trend, and there was no obvious differences between each treatment group. VEGF mRNA expression in TSRDN group was much lower than that in tripterygium group and diosgenin group. CONCLUSION: TSRDN can inhibit angiogenesis in synovial tissue by down regulating expressions of VEGF, Ang-2 and receptor Tie-2.