Four amino acids as serum biomarkers for anti-asthma effects in the ovalbumin-induced asthma mouse model treated with extract of Asparagus cochinchinensis / 한국실험동물학회지

The butanol extract of Asparagus cochinchinensis roots fermented with Weissella cibaria (BAW) effectively prevents inflammation and remodeling of airway in the ovalbumin (OVA)-induced asthma model. To characterize biomarkers that can predict the anti-asthmatic effects induced by BAW treatment, we measured the alteration of endogenous metabolites in the serum of OVA-induced asthma mice after administration of low concentration BAW (BAWLo, 250 mg/kg) and high concentration BAW (BAWHi, 500 mg/kg) using ¹H nuclear magnetic resonance (¹H-NMR) spectral data. The number of immune cells and serum concentration of IgE as well as thickness of the respiratory epithelium and infiltration of inflammatory cells in the airway significantly recovered in the OVA+BAW treated group as compared to the OVA+Vehicle treated group. In the metabolic profile analysis, the pattern recognition showed completely separate clustering of serum analysis parameters between the OVA+Vehicle and OVA+BAW treated groups. Of the total endogenous metabolites, 19 metabolites were upregulated or downregulated in the OVA+Vehicle treated group as compared to the Control treated group. However, only 4 amino acids (alanine, glycine, methionine and tryptophan) were significantly recovered after BAWLo and BAWHi treatment. This study provides the first results pertaining to metabolic changes in the asthma model mice treated with OVA+BAW. Additionally, these findings show that 4 metabolites can be used as one of biomarkers to predict the anti-asthmatic effects.

Dose dependence and durability of the therapeutic effects of Asparagus cochinchinensis fermented extract in an ovalbumin-challenged asthma model / 한국실험동물학회지

The butanol extract of Asparagus cochinchinensis roots fermented with Weissella cibaria (BAfW) significantly suppressed the inflammatory response induced by lipopolysaccharide (LPS) treatment in RAW264.7 cells. To investigate the dose dependence and durability of BAfW on the anti-asthma effects, alterations in key parameters were measured in ovalbumin (OVA)-challenged Balb/c mice treated with the different doses of BAfW at three different time points. The number of immune cells, OVA-specific IgE level, thickness of respiratory epithelium and mucus score decreased significantly in a dose-dependent manner in response to treatment with 125 to 500 mg/kg BAfW (P < 0.05), although the highest level was detected in the 500 mg/kg treated group. Moreover, the decrease in these parameters was maintained from 24 to 48 h in the 500 mg/kg of BAfW treated group. At 72 h, the effects of BAfW on the number of immune cells, OVA-specific IgE level and thickness of respiratory epithelium partially disappeared. Overall, this study provides the first evidence that the anti-asthma effect of BAfW may reach the maximum level in OVA-challenged Balb/c mice treated with 500 mg/kg and that these effects can last for 48 h.

Study on the Purification Technology of Total Saponins from Asparagus cochinchinensis with Macroreticu-lar Resin / 中国药房

OBJECTIVE:To optimize the purification technology of total saponins from Asparagus cochinchinensis with macro-reticular resin. METHODS:Using content of total saponins from A. cochinchinensis as index,single factor test was used to investi-gate the macroreticular resin model,sampling adsorption time,mass concentration of the column,adsorption capacity,volume frac-tion and the amount of elution solvent,elution rate,and optimize the purification technology. And verification test was conducted. RESULTS:HPD-300 macroreticular resin showed strong absorption and desorption property. The optimal purification technology was that sampling adsorption time was 60 min,mass concentration of sample liquid was 0.1 g/mL,adsorption capacity was 120 mL(15 BV),it was eluded with 60% ethanol solution with 3 BV and elution rate was 4 BV/h. In the verification test,the average desorption rate of total saponins was 68.30%(RSD=0.95%,n=3). CONCLUSIONS:Optimized purification technology is sta-ble,feasible,and can easily separate and purify the total saponins from A. cochinchinensis.

Hepatotoxicity and nephrotoxicity of saponin-enriched extract of Asparagus cochinchinensis in ICR mice / 한국실험동물학회지

The inhibitory effects of Asparagus cochinchinensis against inflammatory response induced by lipopolysaccharide (LPS), substance P and phthalic anhydride (PA) treatment were recently reported for some cell lines and animal models. To evaluate the hepatotoxicity and nephrotoxicity of A. cochinchinensis toward the livers and kidneys of ICR mice, alterations in related markers including body weight, organ weight, urine composition, liver pathology and kidney pathology were analyzed in male and female ICR mice after oral administration of 150, 300 and 600 mg/kg body weight/day saponin-enriched extract of A. cochinchinensis (SEAC) for 14 days. The saponin, total flavonoid and total phenol levels were found to be 57.2, 88.5 and 102.1 mg/g in SEAC, respectively, and the scavenging activity of SEAC gradually increased in a dose-dependent manner. Moreover, body and organ weight, clinical phenotypes, urine parameters and mice mortality did not differ between the vehicle and SEAC treated group. Furthermore, no significant alterations were measured in alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), blood urea nitrogen (BUN) and the serum creatinine (Cr) in the SEAC treated group relative to the vehicle treated group. Moreover, the specific pathological features induced by most toxic compounds were not observed upon liver and kidney histological analysis. Overall, the results of the present study suggest that SEAC does not induce any specific toxicity in the livers and kidneys of male and female ICR mice at doses of 600 mg/kg body weight/day.

Therapeutic effect of ethyl acetate extract from Asparagus cochinchinensis on phthalic anhydride-induced skin inflammation / 한국실험동물학회지

Asparagus cochinchinensis has been used to treat various diseases including fever, cough, kidney disease, breast cancer, inflammatory disease and brain disease, while IL-4 cytokine has been considered as key regulator on the skin homeostasis and the predisposition toward allergic skin inflammation. However, few studies have investigated its effects and IL-4 correlation on skin inflammation to date. To quantitatively evaluate the suppressive effects of ethyl acetate extracts of A. cochinchinensis (EaEAC) on phthalic anhydride (PA)-induced skin inflammation and investigate the role of IL-4 during their action mechanism, alterations in general phenotype biomarkers and luciferase-derived signals were measured in IL-4/Luc/CNS-1 transgenic (Tg) mice with PA-induced skin inflammation after treatment with EaEAC for 2 weeks. Key phenotype markers including lymph node weight, immunoglobulin E (IgE) concentration, epidermis thickness and number of infiltrated mast cells were significantly decreased in the PA+EaEAC treated group compared with the PA+Vehicle treated group. In addition, expression of IL-1β and TNF-α was also decreased in the PA+EaEAC cotreated group, compared to PA+Vehicle treated group. Furthermore, a significant decrease in the luciferase signal derived from IL-4 promoter was detected in the abdominal region, submandibular lymph node and mesenteric lymph node of the PA+EaEAC treated group, compared to PA+Vehicle treated group. Taken together, these results suggest that EaEAC treatment could successfully improve PA-induced skin inflammation of IL-4/Luc/CNS-1 Tg mice, and that IL-4 cytokine plays a key role in the therapeutic process of EaEAC.

The New Phytoformula Containing Morus alba, Schizandra sinensis and Asparagus cochinchinensis Inhibits Lung Inflammation in vitro and in vivo

A phytoformula containing the root barks of Morus alba, the fructus of Schizandra sinensis and the roots of Asparagus cochinchinensis (MSA) was prepared as a potential new herbal remedy, and its therapeutic potential for alleviating inflammatory lung conditions was examined. For in vivo evaluation, an animal model of lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice was used. With oral administration of 6 - 60 mg/kg, MSA potently and dose-dependently inhibited bronchitis-like symptoms in acute lung injury induced by intranasal treatment of LPS as judged by the number of cells in the bronchoalveolar lavage fluid (BALF) and histological observation. The inhibitory potency was comparable with that of dexamethasone. For in vitro assay, the effects on the production of proinflammatory molecules in lung epithelial cells and alveolar macrophages were examined. Although MSA inhibited IL-6 production in IL-1β-treated lung epithelial cells (A549) only at a high concentration (300 µg/ml), the formula strongly and concentration-dependently inhibited NO production in LPS-treated alveolar macrophages (MH-S) at 20 - 300 µg/ml. Based on all of these findings, the new phytoformula MSA is suggested to have the potential to control inflammatory lung diseases including bronchitis, at least in part, by inhibiting inducible nitric oxide synthase-catalyzed NO production.

Steroidal saponins from the roots of Asparagus cochinchinensis / 中国天然药物

AIM@#To study the chemical constituents of the roots of Asparagus cochinchinensis (Asparagaceae).@*METHODS@#The compounds were isolated with Diaion HP20, silica gel, and ODS chromatography, and their structures were determined on the basis of chemical methods, HR-ESI-MS, and 1D- and 2D-NMR techniques.@*RESULTS@#Seven compounds were isolated from the n-butanol fraction of the roots of A. cochinchinensis, and their structures were elucidated as (25S)-26-O-β-D-glucopyranosyl-5β-furostan-3β, 22α, 26-triol-12-one-3-O-β-D-glucopyranoside (1), (25S)-26-O-β-D-glucopyranosyl-22α-methoxy-5β-furostan-3β, 26-diol-12-one-3-O-β-D-glucopyranoside (2), (25S)-26-O-β-D-glucopyranosyl-5β-furostan-3β, 22α, 26-triol (3), (25S)-26-O-β-D-glucopyranosyl-5β-furstan-3β, 22α, 26-triol-3-O-β-D-glucopyranoside (4), (25S)-26-O-β-D-glucopyranosyl-5β-furostan-3β, 22α, 26-triol-3-O-α-L-rhamnopyranosyl-(1, 4)-β-D-glucopyranoside (5), (25S)-5β-spirostan-3β-ol-3-O-α-L-rhamnopyranoside (6), and (25S)-5β-spirostan-3β-ol-3-O-β-D-glucopyranoside (7).@*CONCLUSION@#Compounds 1 and 2 were two new furostanol saponins.

Establishment and optimization of ISSR-PCR reaction system in Asparagus cochinchinensis / 中草药

Objective: To establish and optimize ISSR-PCR reaction system for Asparagus cochinchinensis and lay foundation for its genetic diversity research. Methods: The single-factor test and orthogonal design were applied for optimizing seven factors in the ISSR-PCR reaction system including Mg2+, dNTP, primers, Taq DNA polymerase, the template DNA, extension time, and cycle times. Results: The suitable PCR reaction system contained 1.25 mmol/L Mg2+, 320 μmol/L dN TP, 1.2 μmol/L primer, 1.5 U Taq DNA polymerase, and 40 ng template DNA in total 25 μL reaction solution. On this basis, 13 primers were screened with stable amplification and rich polymorphism from 50 ISSR primers. The optimal annealing temperature for ISSR-PCR reaction was proposed by gradient PCR. Conclusion: It is proved that the established and optimized ISSR reaction system would be stable and credible by the germplasm testing result of 17 A. cochinchinensis populations. This would provide the basis for the genetic analysis of A. cochinchinensis.

Studies on chemical constituents of Asparagus cochinchinensis / 第二军医大学学报

Objective: To study the chemical components in the root of Asparagus cochinchinensis (Lour.) Merr. Methods: Chemical components of Asparagus cochinchinensis root were isolated by column chromatography on macroporous resin, Silgel, Sephadex LH-20, ODS, etc. and the structures of the components were identified by physiochemical and spectral analysis, such as MS, 1HNMR, and 13 CNMR. Results: The following 5 components were isolated and identified: diosgenin-3-O-p-β-glu- copyranoside(I), smilagenin(II), 26-O-β-D-glucopyranosyl-furost-3β, 22, 26-triol-3-0-p-D-glucopyranosyl(1→2)-O-β-D-glu- copyranoside(JU) 26-0-β-D-glucopyranosyl-furost-5, 20-en-3β, 2α, 26-triol-3-O-[α-α-rhamnopyranosyl (1[2)]-[α-α-rhamnopyranosyl-(1→4)]-β-D- glucopyranoside(IV), and 26-O-β-D-glucopyranosyl-furost-3β, 26-diol-22-methoxy-3-0-α-L-rhamnopyranosy (1-4)-O-β-D-glucopyranoside (V). Conclusion: All the 5 compounds isolated from Asparagus cochinchinensis in this study is reported for the first time.

Influence of total saponins from Asparagus cochinchinensis on cerebral blood flow and vascular resistance in anesthetized dogs / 第二军医大学学报

Objective: To study the effect of total saponins on cerebral blood flow and vascular resistance in anesthetized dogs. Methods: Thirty hybrid dogs in either sex, with a body weight of (11±1.5) kg, were evenly randomized into 5 groups: negative control group(saline 5 ml/kg,ig), positive control (nimodipine 300 μg/kg, iv), and 3 groups treated with total saponins (low-dose group [1O mg/kg, ig], middle-dose group[30 mg/kg, ig], and high-dose group[60 mg/kg, ig]). The dogs were anesthetized with intravenous pentobarbital sodium (30 mg/kg). The right common carotid artery was exposed to measure the cerebral blood flow, cerebral vascular resistance, blood pressure and heart rate using the MFV-3200 electromagnetic flow meter and MPA-3000 bioelectricity signal-amplifier. Results: Compared with negative control, cerebral blood flow was significantly increased in animals treated with asparagos root saponins (30 and 60 mg/kg, ig) during 5 and 120 min after drug administration (P<0.01). No significant effect on cerebral blood flow and vascular resistance was found in animals treated with asparagus root saponins (10 mg/kg). Conclusion: Asparagus root saponins can increase cerebral blood flow in anesthetized dogs.

Studies on chemical constituents of Asparagus cochinchinensis / 第二军医大学学报

Objective:To study the chemical components in the root of Asparagus cochinchinensis(Lour.)Merr.Methods:Chemical components of Asparagus cochinchinensis root were isolated by column chromatography on macroporous resin,Sil-gel,Sephadex LH-20,ODS,etc.and the structures of the components were identified by physiochemical and spectral analysis,such as MS,1HNMR,and 13CNMR.Results:The following 5 components were isolated and identified:diosgenin-3-O-?-D-glucopyranoside(Ⅰ),smilagenin(Ⅱ),26-O-?-D-glucopyranosyl-furost-3?,22,26-triol-3-O-?-D-glucopyranosyl(1→2)-O-?-D-glucopyranoside(Ⅲ),26-O-?-D-glucopyranosyl-furost-5,20-en-3?,2?,26-triol-3-O-\[?-?-rhamnopyranosyl(1→2)\]-\[?-?-rhamnopyranosyl-(1→4)\]-?-D-glucopyranoside(Ⅳ),and 26-O-?-D-glucopyranosyl-furost-3?,26-diol-22-methoxy-3-O-?-L-rhamnopyranosy(1→4)-O-?-D-glucopyranoside(Ⅴ).Conclusion:All the 5 compounds isolated from Asparagus cochinchinensis in this study is reported for the first time.

Influence of total saponins from Asparagus cochinchinensis on cerebral blood flow and vascular resistance in anesthetized dogs / 第二军医大学学报

Objective:To study the effect of total saponins on cerebral blood flow and vascular resistance in anesthetized dogs.Methods:Thirty hybrid dogs in either sex,with a body weight of(11?1.5)kg,were evenly randomized into 5 groups: negative control group(saline 5 ml/kg,ig),positive control(nimodipine 300?g/kg,iv),and 3 groups treated with total saponins (low-dose group[10 mg/kg,ig],middle-dose group[30 mg/kg,ig],and high-dose group[60 mg/kg,ig]).The dogs were anes- thetized with intravenous pentobarbital sodium(30 mg/kg).The right common carotid artery was exposed to measure the cere- bral blood flow,cerebral vascular resistance,blood pressure and heart rate using the MFV-3200 electromagnetic flow meter and MPA-3000 bioelectricity signal-amplifier.Results:Compared with negative control,cerebral blood flow was significantly in- creased in animals treated with asparagus root saponins(30 and 60 mg/kg,ig)during 5 and 120 min after drug administration (P

HPLC-ELSD in determination of AR-Ⅰ content in total furostanol saponins of Asparagus cochinchinensis / 第二军医大学学报

Objective:To determine AR-Ⅰ content in total furostanol saponins of Asparagus cochinchinensis.Methods: High performance liquid chromatography(HPLC) was used and the condition was: Diamonsil C_(18) column(5 mm?250 mm,5 ?m);mobile phase:(CH_(3)CN-H_(2)O(36∶64);) flow rate: 0.8 ml/min;injector volume:20 ?l;colum temperature: room temperature.Evaporative light scattering detector(ELSD) condition was: drift tube temperature: 40℃;and nebulizer gas pressure(N_(2)): 2.0?10~(5) Pa.Results: The linearity was good when AR-I was within the range of 0.094-0.940 0 g?L~(-1)(r=0.999 2).The test had higher sensitivity and good stability.The average recovery was 101.3%(n=6)and RSD was 2.36%.Conclusion: HPLC-ELSD is practical and reliable for the determination of the furostanol saponins of Asparagus cochinchinensis.