Roots Of Panax Notoginseng Research Articles

Notoginsenoside R1 protects boar sperm during liquid storage at 17°C.

Reactive oxygen species (ROS) damage mammalian sperm during liquid storage. Notoginsenoside R1 (NR1) is a compound isolated from the roots of Panax notoginseng; it has powerful ROS-scavenging activities. This work hypothesized that the antioxidant capacity of NR1 could improve boar sperm quality and fertility during liquid storage. During liquid storage at 17°C, the supplementation of semen extender with NR1 (50μM) significantly improved sperm motility, membrane integrity and acrosome integrity after 5days of preservation. NR1 treatment also reduced ROS and lipid peroxidation (LPO) levels at day 5 (p<0.05). Higher glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) levels and sperm-zona pellucida binding capacity were observed in the 50μM NR1 group than those in the control group at day 7 (p<0.05). Importantly, statistical analysis of the fertility of 200 sows indicated that addition of NR1 to the extender improved the fertility parameters of boar spermatozoa during liquid storage at 17°C (p<0.05). These results demonstrate the practical feasibility of using 50μM NR1 as an antioxidant in boar extender during liquid storage at 17°C, which is beneficial to both spermatozoa quality and fertility.

Effect of pulsed vacuum drying on drying kinetics and quality of roots of Panax notoginseng (Burk.) F. H. Chen (Araliaceae)

In order to explore the feasibility to enhance drying kinetics, the effects of pulsed vacuum drying (PVD) under different temperatures (55, 60, 65, 70, and 75 °C) and pulsed vacuum ratio (PVR) (3:16, 6:16, 9:16, 12:16, 9:10, 9:13, and 9:19 min/min) on the drying kinetics, energy consumption, and quality attributes of Panax notoginseng roots in terms of color parameters (L*, a*, b*, and △E), the contents of ginsenosides (R1, Rg1, Re, Rd, Rb1), and Panax notoginseng Saponins (PNS) are evaluated. The results were compared with those obtained under hot air drying (HAD) conducted at the temperature of 60 °C, relative humidity of 12%, and air flow rate of 0.3 m/s. PVD at 75 °C and PVR of 9:16 min/min resulted in the shortest drying time, i.e., 26.6 h, while PVD at the temperature of 65 °C and PVR of 9:13 resulted in the highest PNS content, i.e., 9.03%. Among different drying conditions, the temperature of 65 °C and PVR of 9:16 min/min are recommended as the most suitable ones for drying of whole Panax notoginseng roots in terms of drying time, energy consumption and quality of final product.

A magnetic multi-walled carbon nanotube preparative method for analyzing asymmetric carbon, phosphorus and sulfur atoms of chiral pesticide residues in Chinese herbals by chiral liquid chromatography-quadrupole/linear ion trap mass spectrometry determination

An analytical method for the determination of asymmetric carbon, phosphorus and sulfur atoms in chiral pesticide residues by magnetic multi-walled carbon nanotube sample pretreatment combined with chiral ultra-performance liquid chromatography/quadrupole/linear ion trap mass spectrometry (UPLC-MS/Qtrap) was developed and applied to chiral pesticide residues analysis in Chinese herbals. Eleven different chiral pesticides were found, and 36.4% were positive in Chinese herbals. Three plants containing detectable pesticide residues were observed in Dendrobium nobile, Panax notoginseng flowers and honeysuckle, in the order of decreasing detected concentration. High detection frequencies of 26.1% for (R/S)-(±)-difenoconazole and 14.5% for (R/S)-(±)-metalaxyl and (R/S)-(±)-propiconazole were observed, the residual amount for (R/S)-(±)-difenoconazole, (R/S)-(±)-metalaxyl and (R/S)-(±)-propiconazole were 0.32 ~ 2.5 mg/kg, 0.022 ~ 0.23 mg/kg, 0.62 ~ 3.21 mg/kg respectively. The EF value of (R/S)-(±)-difenoconazole was 0.506 ± 0.046. The EF value of (R/S)-(±)-metalaxyl was lower than 0.5 in Dendrobium nobile, Panax notoginseng flowers, Panax notoginseng roots and hawthorn. The EF of (R/S)-(±)-propiconazole was not significantly enantioselective in honeysuckle and Panax notoginseng flowers. The enantioselectivity of various pesticide residues in different plants cannot be predicted from our existing knowledge and may closely depend on plant growth, environmental conditions or molecular structure.

Magnesium reduces cadmium accumulation by decreasing the nitrate reductase-mediated nitric oxide production in Panax notoginseng roots

Panax notoginseng is a traditional medicinal herb in China. However, the high capacity of its roots to accumulate cadmium (Cd) poses a potential risk to human health. Our previous study showed that nitrate reductase (NR)-dependent nitric oxide (NO) production promoted Cd accumulation in P. notoginseng root cell walls. In this study, the role of Mg in the regulation of NO production and Cd accumulation in P. notoginseng roots was characterized. Exposure of P. notoginseng roots to increasing concentrations of Cd resulted in a linear increase in NO production. The application of 2 mM Mg for 24 h significantly alleviated Cd-induced NO production and Cd accumulation in roots, which coincided with a significant decrease in the NR activity. Western analysis suggested that Mg increased the interaction between the 14-3-3 protein and NR, which might have been a reason for the Mg-mediated decrease in NR activity and NO production under Cd stress. These results suggested that Mg-mediated alleviation of Cd-induced NO production and Cd accumulation is achieved by enhancement of the interaction between the 14-3-3 protein and NR in P. notoginseng roots.

Comparison of intramuscular and intravenous pharmacokinetics of ginsenosides in humans after dosing XueShuanTong, a lyophilized extract of Panax notoginseng roots

Ethnopharmacological relevanceMany bioactive constituents of Chinese herbal medicines have poor oral bioavailability. Besides oral administration, herbal medicines in China are also prepared for parenteral administration. Unlike for intravenous route, little is known about the intramuscular pharmacokinetics of herbal compounds. To facilitate rational use of herbal medicine, it is important to better understand such intramuscular pharmacokinetics. Aim of the studyBioactive constituents of XueShuanTong (a lyophilized extract of Panax notoginseng roots, extensively used in treatment of ischemic heart and cerebrovascular diseases) predominantly comprise ginsenosides Rb1 and Rd of 20(S)-protopanaxadiol-type and ginsenosides Rg1, and Re, and notoginsenoside R1 of 20(S)-protopanaxatriol-type; these saponins are poorly absorbed from the gastrointestinal tract. This study aimed to compare intramuscular and intravenous pharmacokinetics of these ginsenosides after dosing XueShuanTong. MethodsPharmacokinetics of ginsenosides was assessed in human volunteers receiving an intramuscular injection or 1.5-h intravenous infusion of XueShuanTong, both at 150 mg/person, and the plasma and urine samples were analyzed by liquid chromatography/mass spectrometry. ResultsLike after intravenous administration, the unchanged saponins were the major circulating forms after intramuscular administration, while their metabolites were poorly detected. These ginsenosides exhibited intramuscular bioavailability of 100%–112%, relative to the respective intravenous data. Similar to that after intravenous infusion, the 20(S)-protopanaxadiol-type ginsenosides after the intramuscular injection exhibited notably longer terminal half-lives (46–106 h) than the 20(S)-protopanaxatriol-type ginsenosides (1.1–1.4 h). ConclusionsIntramuscular route might be an effective alternative to intravenous route for XueShuanTong, from the pharmacokinetic perspective.

&lt;p&gt;Focus on Notoginsenoside R1 in Metabolism and Prevention Against Human Diseases&lt;/p&gt;

Notoginsenoside (NG)-R1 is one of the main bioactive compounds from Panax notoginseng (PN) root, which is well known in the prescription for mediating the micro-circulatory hemostasis in human. In this article, we mainly discuss NG-R1 in metabolism and the biological activities, including cardiovascular protection, neuro-protection, anti-diabetes, liver protection, gastrointestinal protection, lung protection, bone metabolism regulation, renal protection, and anti-cancer. The metabolites produced by deglycosylation of NG-R1 exhibit higher permeability and bioavailability. It has been extensively verified that NG-R1 may ameliorate ischemia-reperfusion (IR)-induced injury in cardiovascular and neuronal systems mainly by upregulating the activity of estrogen receptor α-dependent phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) and nuclear factor erythroid-2-related factor 2 (NRF2) pathways and downregulating nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) pathways. However, no specific targets for NG-R1 have been identified. Expectedly, NG-R1 has been used as a main bioactive compound in many Traditional Chinese Medicines clinically, such as Xuesaitong, Naodesheng, XueShuanTong, ShenMai, and QSYQ. These suggest that NG-R1 exhibits a significant potency in drug development.

Stem-leaves of Panax as a rich and sustainable source of less-polar ginsenosides: comparison of ginsenosides from Panax ginseng, American ginseng and Panax notoginseng prepared by heating and acid treatment

BackgroundGinsenosides, which have strong biological activities, can be divided into polar or less-polar ginsenosides. MethodsThis study evaluated the phytochemical diversity of the saponins in Panax ginseng (PG) root, American ginseng (AG) root, and Panax notoginseng (NG) root; the stem-leaves from Panax ginseng (SPG) root, American ginseng (SAG) root, and Panax notoginseng (SNG) root as well as the saponins obtained following heating and acidification [transformed Panax ginseng (TPG), transformed American ginseng (TAG), transformed Panax notoginseng (TNG), transformed stem-leaves from Panax ginseng (TSPG), transformed stem-leaves from American ginseng (TSAG), and transformed stem-leaves from Panax notoginseng (TSNG)]. The diversity was determined through the simultaneous quantification of the 16 major ginsenosides. ResultsThe content of ginsenosides in NG was found to be higher than those in AG and PG, and the content in SPG was greater than those in SNG and SAG. After transformation, the contents of polar ginsenosides in the raw saponins decreased, and contents of less-polar compounds increased. TNG had the highest levels of ginsenosides, which is consistent with the transformation of ginseng root. The contents of saponins in the stem-leaves were higher than those in the roots. The transformation rate of SNG was higher than those of the other samples, and the loss ratios of total ginsenosides from NG (6%) and SNG (4%) were the lowest among the tested materials. In addition to the conversion temperature, time, and pH, the crude protein content also affects the conversion to rare saponins. The proteins in Panax notoginseng allowed the highest conversion rate. ConclusionThus, the industrial preparation of less-polar ginsenosides from SNG is more efficient and cheaper.

The transcriptome variations of Panaxnotoginseng roots treated with different forms of nitrogen fertilizers

BackgroundThe sensitivity of plants to ammonia is a worldwide problem that limits crop production. Excessive use of ammonium as the sole nitrogen source results in morphological and physiological disorders, and retarded plant growth.ResultsIn this study we found that the root growth of Panax notoginseng was inhibited when only adding ammonium nitrogen fertilizer, but the supplement of nitrate fertilizer recovered the integrity, activity and growth of root. Twelve RNA-seq profiles in four sample groups were produced and analyzed to identify deregulated genes in samples with different treatments. In comparisons to NH{~}_{4}^{+} treated samples, ACLA-3 gene is up-regulated in samples treated with NO{~}_{3}^{-} and with both NH_{4}^{+} and NO{~}_{3}^{-}, which is further validated by qRT-PCR in another set of samples. Subsequently, we show that the some key metabolites in the TCA cycle are also significantly enhanced when introducing NO{~}_{3}^{-}. These potentially enhance the integrity and recover the growth of Panax notoginseng roots.ConclusionThese results suggest that the activated TCA cycle, as demonstrated by up-regulation of ACLA-3 and several key metabolites in this cycle, contributes to the increased Panax notoginseng root yield when applying both ammonium and nitrate fertilizer.

In Vitro Anti-hepatoma Activities of Notoginsenoside R1 Through Downregulation of Tumor Promoter miR-21.

Notoginsenoside R1 (NG-R1) is the predominant active ingredient and a novel triterpene saponin compound extracted from the roots of Panax notoginseng. To date, to the best of our knowledge, there are no previous studies concerning the effect of NG-R1 on hepatocellular carcinoma (HCC). To investigate the effects of NG-R1 on HCC cell growth, apoptosis, and invasion and to explore the underlying mechanisms. Cell viability and lactate dehydrogenase (LDH) release were evaluated by cell counting kit-8 and LDH assay, respectively. Apoptosis was assessed using flow cytometry analysis and caspase-3/7 activity assay. Cell invasion was detected by Transwell invasion assay and western blot analysis of matrix metallopeptidase (MMP)-2 and MMP-9. The effects of NG-R1 on miR-21 expression and phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) pathway were examined by qRT-PCR and western blot, respectively. NG-R1 inhibited the viability, increased LDH release and caspase-3/7 activity, induced apoptosis, and suppressed invasion in HCC cells. NG-R1 reduced miR-21 expression in HCC cells. miR-21 overexpression significantly attenuated the effects of NG-R1 on the viability, LDH release, apoptosis, caspase-3/7 activity, and invasion of HCC cells. We further demonstrated that NG-R1 inhibited the activation of the PI3K/Akt pathway in HCC cells, which was abolished by miR-21 overexpression. NG-R1 exerted anti-hepatoma activity through inactivation of the PI3K/Akt pathway by downregulating miR-21, contributing to further understanding of the anti-tumor activities of NG-R1 in HCC.

Intravenous formulation of Panax notoginseng root extract: human pharmacokinetics of ginsenosides and potential for perpetrating drug interactions.

XueShuanTong, a lyophilized extract of Panax notoginseng roots (Sanqi) for intravenous administration, is extensively used as add-on therapy inthe treatment of ischemic heart and cerebrovascular diseases and comprises therapeutically active ginsenosides. Potential for XueShuanTong-drug interactions was determined; the investigation focused on cytochrome P450 (CYP)3A induction and organic anion-transporting polypeptide (OATP)1B inhibition. Ginsenosides considerably bioavailable for drug interactions were identified by dosing XueShuanTong in human subjects and their interaction-related pharmacokinetics were determined. The CYP3A induction potential was determined by repeatedly dosing XueShuanTong for 15 days in human subjects and by treating cryopreserved human hepatocytes with circulating ginsenosides; midazolam served as a probe substrate. Joint inhibition of OATP1B by XueShuanTong ginsenosides was assessed in vitro, and the data were processed using the Chou-Talalay method. Samples were analyzed by liquid chromatography/mass spectrometry. Ginsenosides Rb1, Rd, and Rg1 and notoginsenoside R1 were the major circulating XueShuanTong compounds; their interaction-related pharmacokinetics comprised compound dose-dependent levels of systemic exposure and, for ginsenosides Rb1 and Rd, long terminal half-lives (32‒57 and 58‒307 h, respectively) and low unbound fractions in plasma (0.8%‒2.9% and 0.4%‒3.0%, respectively). Dosing XueShuanTong did not induce CYP3A. Based on the pharmacokinetics and inhibitory potency of the ginsenosides, XueShuanTong was predicted to have high potential for OATP1B3-mediated drug interactions (attributed chiefly to ginsenoside Rb1) suggesting the need for further model-based determination of the interaction potential for XueShuanTong and, if necessary, a clinical drug interaction study. Increased awareness of ginsenosides' pharmacokinetics and XueShuanTong-drug interaction potential will help ensure thesafe use of XueShuanTong and coadministered synthetic drugs.

Toxicological Evaluation of a Mixture of Astragalus membranaceus and Panax notoginseng Root Extracts (InnoSlim®).

Astragalus spp. and Panax spp. have a long history of traditional human use. A blend, InnoSlim®, of highly purified and fractionated root extracts from Astragalus membranaceus and Panax notoginseng has now been developed for human consumption; however, the unique constituent content of this blend has not been specifically evaluated with respect to safety. Therefore, the toxicological potential of the blend was formally investigated in a series of studies—genetic toxicity was evaluated in a bacterial reverse mutation test followed by an in vivo mammalian micronucleus test, and general toxicity was evaluated in a 28-day repeated-dose oral toxicity study in rats. No evidence of mutagenicity was observed in the bacterial tester strains used, and no evidence of in vivo chromosomal damage resulting in increased frequency of micronucleated cells was observed in male Crl:NMRI BR mice. No mortality or toxic effects were observed, and no target organs were identified, in male and female Han:WIST rats exposed to 0, 400, 800, or 1200 mg/kg bw/day of the blend by gavage for 28 consecutive days. The highest dose—1200 mg/kg bw/day—was determined to be the NOAEL. Based on these results, extrapolation towards a safe human consumption level can be explored.

Anti-inflammatory and Cytotoxic Triterpenes from the Rot Roots of Panax notoginseng

Four new protopanaxatriol-type triterpenes (1–2) and glucosides (3–4), were isolated from the rot roots of Panax notoginseng (Burk.) Chen, along with four known ones (5–8). Their structures were elucidated on the basis of extensive spectroscopic analysis (HRESIMS, NMR, UV, IR, and OR) and acidic hydrolysis. The possible transformation pathway of these compounds were also speculated from ginsenoside Rg1. Compound 1, with a unique α,β-unsaturated ketene in its side chain, showed significant inhibitory effects against NO production on Murine macrophage cells (IC50 = 4.12 ± 0.20 μM) and comparable cytotoxicities against five human cancer cell lines (myeloid leukemia HL-60, lung cancer A-549 cells, hepatocellular carcinoma SMMC7721, breast cancer MCF-7, and colon cancer SW480) to positive control, cisplatin (DDP).Graphical

Notoginseng Saponin Rg1 Prevents Cognitive Impairment through Modulating APP Processing in Aβ1-42-injected Rats.

With the intensification of the aging process of the world, Alzheimer's disease (AD), which is the main type of senile dementia, has become a primary problem in the present society. Lots of strategies have been used to prevent and treat AD in animal models and clinical trials, but most of them ended in failure. Panax notoginseng saponins (PNS) contain a variety of monomer compositions which have been separated and identified. Among of the monomer compositions, notoginseng saponin Rg1 (Rg1) accounts for 20% of the cultivation of panax notoginseng roots. And now PNS have been reported to be widely used to treat cardio-cerebrovascular diseases and have neuroprotective effects to restrain the β-amyloid peptide (Aβ)25-35-mediated apoptosis. Moreover, it is reported that PNS could accelerate the growth of nerve cells, increase the length of axons and promote synaptic plasticity. Whether Rg1 can ameliorate the cognitive impairment and the underlying mechanism has not been elucidated. To study the preventive effect of Rg1 on cognitive impairment and the possible mechanism, we established the cognitive impairment model in rats through Aβ1-42 (2.6 µg/µL, 5 µL) injection and then treated the rats with Rg1 (25, 50 and 100 mg/kg) administered intragastrically for 4 weeks. We observed that Aβ1-42 could induce spatial learning and memory deficits in rats. Simultaneously, Aβ1-42 injection also resulted in the reduced neuron number in cornuammonis 1 (CA1) and dentate gyrus (DG) of hippocampus, as well as the increased level of hyperphosphorylated β-amyloid precursor protein (APP) at Thr668 site with up-regulation of β-APP cleaving enzyme 1 (BACE1) and presenilin 1 (PS1) and down-regulation of a disintegrin and metalloprotease domain-containing protein 10 (ADAM10) and insulin-degrading enzyme (IDE). Administration of Rg1 effectively rescued the cognitive impairment and neuronal loss, and inhibited the β-secretase processing of APP through reducing APP-Thr668 phosphorylation and BACE1/PS1 expression, and increasing the expression of ADAM10 and IDE. We concluded that Rg1 might have neuroprotective effects and could promote learning and memory ability, which might be a viable candidate in AD therapy probably through reducing the generation of Aβ and increasing the degradation of Aβ.

Antifungal Effect of Essential Oils from Five Kinds of Rutaceae Plants - Avoiding Pesticide Residue and Resistance.

Panax notoginseng root is a traditional Chinese herb, of which the yield and quality have been seriously affected by microorganisms, and is commonly used to treat various kinds of bleeding. In this experiment, the effects of the antifungal properties of essential oils (EOs) from five kinds of Rutaceae plants on the growth of three kinds of pathogens were studied to develop natural, environmentally friendly antifungal agents. Citrus medica EO was found to have stronger inhibitory effects on the growth of pathogenic fungi in vitro than other EOs with the Oxford cup method, of which the chemical composition was further investigated by GC/MS. The major components were d-limonene (22.79 %) and γ-terpinene (9.71 %). The antifungal activities were evaluated by MIC and FIC assays. In these assays, C. medica EO, d-limonene and γ-terpinene were effective against three pathogens of P. notoginseng with MIC values ranging from 0.12 to 12.05 mg/mL. The association between hymexazol and C. medica EO showed a high synergistic effect with lower FIC index values (FICi=0.31-2.00). Furthermore, C. medica EO was further assessed in P. notoginseng planted in a continuous cropping soil (CCS) and was found to reduce the disease incidence and disease severity compared with P. notoginseng planted in CCS only without EO addition. This finding suggested that C. medica EO has potential as a natural environmentally antifungal agent against pathogens of P. notoginseng, ensuring its safety.

New hydroperoxylated and 20,24-epoxylated dammarane triterpenes from the rot roots of Panax notoginseng

BackgroundRoot rot is a serious destructive disease of Panax notoginseng, a famous cultivated araliaceous herb called Sanqi or Tianqi in Southwest China. MethodsThe chemical substances of Sanqi rot roots were explored by chromatographic techniques. MS, 1D/2D-NMR, and single crystal X-ray diffraction were applied to determine the structures. Murine macrophage RAW264.7 and five human cancer cell lines were used separately for evaluating the antiinflammatory and cytotoxic activities. Results and ConclusionThirty dammarane-type triterpenes and saponins were isolated from the rot roots of P. notoginseng. Among them, seven triterpenes, namely, 20(S)-dammar-25-ene-24(S)-hydroperoxyl-3β,6α,12β,20-tetrol (1), 20(S)-dammar-3-oxo-23-ene-25-hydroperoxyl-6α,12β,20-triol (2), 20(S)-dammar-12-oxo-23-ene-25-hydroperoxyl-3β,6α,20-triol (3), 20(S)-dammar-3-oxo-23-ene-25-hydroperoxyl-12β,20-diol (4), 20(S),24(R)-epoxy-3,4-seco-dammar-25-hydroxy-12-one-3-oic acid (5), 20(S),24(R)-epoxy-3,4-seco-dammar-25-hydroxy-12-one-3-oic acid methyl ester (6), and 6α-hydroxy-22,23,24,25,26,27-hexanordammar-3,12,20-trione (7), are new compounds. In addition, 12 known ones (12–16 and 19–25) were reported in Sanqi for the first time. The new Compound 1 showed comparable antiinflammatory activity on inhibition of NO production to the positive control, whereas the known compounds 9, 12, 13, and 16 displayed moderate cytotoxicities against five human cancer cell lines. The results will provide scientific basis for understanding the chemical constituents of Sanqi rot roots and new candidates for searching antiinflammatory and antitumor agents.

Antioxidant and anti-aging activities and structural elucidation of polysaccharides from Panax notoginseng root

Abstract Two novel polysaccharides, MRP5 and MRP5A, were obtained from the root of Panax notoginseng using column chromatography. Structure characterization suggested that MRP5A, with an average molecular weight of 11.38 × 104 Da, was composed of rhamnose (7.7%), arabinose (6.6%), glucose (69.8%) and galactose (15.9%). MRP5 had an average molecular weight of 9.16 × 104 Da and consisted of rhamnose (3.6%), arabinose (14.5%), glucose (51.2%) and galactose (29.7%). The results of methylation analysis and one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) spectra reveal that MRP5 contains repeating units of →3)-β-Glcp-(1→, →6)-β-Glcp-(1→, →3, 6)-Glcp-(1→, β-Glcp-(1→, →3)-β-Galp-(1→, →3, 6)-β-Galp-(1→, →3)-α-Rhap-(1→, →3)-α-Araf-(1→, and α-Araf-(1→ residues. MRP5 and MRP5A exhibited relatively low antioxidant abilities in vitro, but strongly protective effects against oxidative stress and lifespan extension effects in C. elegans. Overall, the present study indicated that MRP5 could be further investigated as a functional food for promoting healthspan.

Notoginsenoside R1 up-regulates microRNA-132 to protect human lung fibroblast MRC-5 cells from lipopolysaccharide-caused injury

BackgroundPneumonia is a common lung disease in children with high fatality rate. Notoginsenoside R1 (NGR1) is the main active component extracted from the roots of Panax notoginseng (Burk.) F.H. Chen (Araliaceae). Here, we carefully explored the potential anti-inflammatory and protective effects of NGR1 on lipopolysaccharide (LPS)-induced lung fibroblast MRC-5 cell injury. MethodsViability and apoptosis of MRC-5 cells after different treatment or transfection were respectively assessed using CCK-8 assay and Annexin V-FITC/PI staining. The expression levels of microRNA-132 (miR-132), IL-1β, IL-6 and TNF-α in MRC-5 cells were measured using qRT-PCR. MicroRNA transfection was conducted to reduce the expression level of miR-132. Western blotting was used to analyze the protein expression levels of key factors involving in cell proliferation, apoptosis, NF-κB pathway and JNK pathway. ResultsLPS treatment caused MRC-5 cell proliferation inhibition, apoptosis and over-production of inflammatory cytokines. NGR1 treatment had no significant effects on MRC-5 cell proliferation, apoptosis and production of inflammatory cytokines, but protected MRC-5 cells from LPS-caused cell proliferation inhibition, apoptosis and over-production of inflammatory cytokines. In addition, NGR1 increased the expression level of miR-132 in MRC-5 cells. Knockdown of miR-132 reversed the protective effects of NGR1 on LPS-treated MRC-5 cells. Furthermore, NGR1 attenuated LPS-activated NF-κB and JNK pathways in MRC-5 cells via up-regulation of miR-132. ConclusionThis research confirmed the protective roles of NGR1 in lung fibroblast cell inflammatory injury. NGR1 protected MRC-5 cells from LPS-caused inflammatory injury through up-regulating miR-132 and then inactivating NF-κB and JNK pathways.

Transcriptome analysis of 1- and 3-year-old Panax notoginseng roots and functional characterization of saponin biosynthetic genes DS and CYP716A47-like.

Transcriptome analysis revealed high expression of saponin biosynthetic genes may account for highly accumulated saponins in 3-year-old Panax notoginseng roots and DS and CYP716A47 - like were functionally verified by transgenic tobacco. Panax notoginseng is a well-known traditional medical herb that contains bioactive compounds known as saponins. Three major dammarene-type triterpene saponins including R1, Rb1, and Rg1 were found to be highly accumulated in the roots of 3-year-old plants when compared to those of 1-year-old plants. However, the underlying cellular mechanism is poorly understood. In this study, transcriptome analysis revealed that most genes involved in saponin biosynthesis in P. notoginseng roots augmented during their growth periods. The analysis of the KEGG pathway indicated that the primary metabolism, cell growth, and differentiation were less active in the roots of 3-year-old plant; however, secondary metabolisms were enhanced, thus providing molecular evidence for the harvesting of P. notoginseng roots in the 3rd year of growth. Furthermore, the functional role of DS and CYP716A47-like, two of the candidate genes involved in saponin biosynthesis isolated from P. notoginseng, were verified via overexpression in cultivated tobacco. Approximately, 0.325µgg-1 of dammarenediol-II and 0.320µgg-1 of protopanaxadiol were recorded in the dry leaves of transgenic tobacco overexpressed with DS and both DS and CYP716A47-like, respectively. This study provides insights into the molecular mechanisms for saponin accumulation in P. notoginseng roots during its growth period and paves a promising way to produce dammarenediol-II and protopanaxadiol via transgenic techniques.

Identification of anti-inflammatory components of raw and steamed Panax notoginseng root by analyses of spectrum-effect relationship.

To identify bioactive components of Panax notoginseng (PN) roots in raw and steamed forms, chemometrics analyses including bivariate correlation analysis and partial least squares regression were used to establish the relationships between the chromatograms and anti-inflammatory effect of PN samples. The chemical fingerprints of PN were obtained by high performance liquid chromatography. The anti-inflammatory activity was evaluated by the TNF-α and IL-6 inhibiting test. According to the results, raw PN (RPN) displayed a stronger anti-inflammatory effect compared with steamed PN (SPN) samples. Chemometrics analyses showed that notoginsenoside R1, ginsenosides Rg1, Re and Rb1 were the major anti-inflammatory components of RPN, whereas ginsenosides 20(S)-Rg3 was the active one of SPN treating inflammation. The pharmacologic verification was consistent with the predicted results. In addition, the contents of anti-inflammatory components in RPN were higher than those in SPN, which might be the reason that RPN exhibited stronger effect on the inflammation induced by lipopolysaccharide in RAW264.7 cells. These findings could be beneficial for choosing differential markers for the quality control of RPN and SPN.

Bioactive Proteins in Panax notoginseng Roots and Other Panax Species.

The genus Panax consists of a group of prized medicinal herbs. Major members of the Panax genus include P. ginseng, P. notoginseng, P. quinquefolius, and P. vietnamensis. They possess various bioactive constituents such as ginsenosides, saponins, polysaccharides and proteins. Many of them were reported to show beneficial effects on human health. Ginsenosides and saponins of ginsengs caught the sight of most researchers. Precise investigations revealed their roles on improvement of the functioning of the nervous system, cardiovascular system, and other functions. In contrast, our knowledge of the bioactive Panax proteins is relatively limited. A number of proteins from P. ginseng, the most valuable member of Panax species, have been investigated and proved to be beneficial to our body. Meanwhile, a few bioactive P. notoginseng proteins, such as ribonucleases and antifungal proteins, have been characterized and reported. We summarize herein the proteins present in P. notoginseng that have been identified, and try to compare them with those from other Panax species with a similar structure or bioactivity, and conclude whether the proteins in P. notoginseng have any distinctive features.