Drug markets revisits herbal medicines, as historical usages address their therapeutic efficacies with less adverse effects. Moreover, herbal medicines save both cost and time in development. SH003, a modified version of traditional herbal medicine extracted from Astragalus membranaceus (Am), Angelica gigas (Ag), and Trichosanthes Kirilowii Maximowicz (Tk) with 1:1:1 ratio (w/w) has been revealed to inhibit tumor growth and metastasis on highly metastatic breast cancer cells, both in vivo and in vitro with no toxicity. Meanwhile, autophagy is imperative for maintenance cellular homeostasis, thereby playing critical roles in cancer progression. Inhibition of autophagy by pharmacological agents induces apoptotic cell death in cancer cells, resulting in cancer treatment. In this study, we demonstrate that SH003-induced autophagy via inhibiting STAT3 and mTOR results in an induction of lysosomal p62/SQSTM1 accumulation-mediated reactive oxygen species (ROS) generation and attenuates tumor growth. SH003 induced autophagosome and autolysosome formation by inhibiting activation of STAT3- and mTOR-mediated signaling pathways. However, SH003 blocked autophagy-mediated p62/SQSTM1 degradation through reducing of lysosomal proteases, Cathepsins, resulting in accumulation of p62/SQSTM1 in the lysosome. The accumulation of p62/SQSTM1 caused the increase of ROS, which resulted in the induction of apoptotic cell death. Therefore, we conclude that SH003 suppresses breast cancer growth by inducing autophagy. In addition, SH003-induced p62/SQSTM1 could function as an important mediator for ROS generation-dependent cell death suggesting that SH003 may be useful for treating breast cancer.

Cardiovascular disease (CVD), the number one cause of death worldwide, has always been the focus of clinical and scientific research. Due to the high number of deaths each year, it is essential to find alternative therapies that are safe and effective with minimal side effects. Traditional Chinese medicine (TCM) has a long history of significant impact on the treatment of CVDs. The mode of action of natural active ingredients of drugs and the development of new drugs are currently hot topics in research on TCM. Astragalus membranaceus is a commonly used Chinese medicinal herb. Previous studies have shown that Astragalus membranaceus has anti-tumor properties and can regulate metabolism, enhance immunity, and strengthen the heart. Astragaloside IV (AS-IV) is the active ingredient of Astragalus membranaceus, which has a prominent role in cardiovascular diseases. AS-IV can protect against ischemic and hypoxic myocardial cell injury, inhibit myocardial hypertrophy and myocardial fibrosis, enhance myocardial contractility, improve diastolic dysfunction, alleviate vascular endothelial dysfunction, and promote angiogenesis. It can also regulate blood glucose and blood lipid levels and reduce the risk of cardiovascular diseases. In this paper, the mechanism of AS-IV intervention in cardiovascular diseases in recent years is reviewed in order to provide a reference for future research and new drug development.

Keywords: Astragalus membranaceus; astragaloside IV; cardiomyocytes; cardiovascular diseases; traditional Chinese medicine.

By aging, male fertility and kidney function decline. Therefore, the investigation of health span-extending agents becomes more urgent to overcome aging-induced infertility and kidney dysfunction. The current research was undertaken to investigate the antiaging efficacy of Astragalus membranaceus telomerase activator-65 (Ta-65) and pomegranate supplements. Forty male Wistar rats were divided into young rats, aged rats, aged rats treated with Ta-65 (500mg/kg/day), and aged rats treated with pomegranate (250mg/kg/day). Testosterone, FSH, LH, and kidney functions were measured in serum. Sperm analysis as well as testicular histological examination was performed. Aging caused an imbalance in male sex hormones resulting in sperm abnormality and reductions in the sperm count and motility. Elevations in serum creatinine, uric acid, sodium, and potassium were reported in aged rats. Treatment with Ta-65 or pomegranate effectively ameliorated all the deteriorations induced by normal aging in male fertility and renal function. Ta-65 and pomegranate possessed strong antiaging activity by alleviating aging-induced male infertility through reestablishing the hormonal balance and testis architecture. They also alleviated the kidney dysfunction. On comparing Ta-65 with pomegranate, the improvement in FSH, LH, and sperm abnormalities caused by Ta-65 was much better than that caused by pomegranate.

Keywords: FSH; Infertility; LH; kidney dysfunction; sperm analysis; testosterone.

The purpose of this study was to investigate whether adding a Chinese herbal hedicine (CHM, including astragalus membranaceus, codonopsis pilosula, poria cocos, bighead atractylodes rhizome, fructus crataegi, folium epimedii, radix saposhnikoviae, and pericarpium citri reticulatae) diet has effects on lipid metabolism in the liver of the broiler. Six-hundred-thirty Hetian broilers were divided into 3 groups including base diet (control), 0.3%, or 1% CHM diets, for 130 days. The results showed that CHM treatment significantly increased the slaughter performance. CHM treatment decreased plasma triglyceride (TG) and nonesterified fatty acid (NEFA) concentrations; up-regulated the expression of peroxisome proliferator activated receptor (PPAR), the transcription factor sterol regulatory element binding protein-1c (SREBP-1c) and its downstream proteins involved in NEFA oxidation including carnitine palmitoyltransferase-1, liver fatty acid binding protein, and PPARα; and it significantly decreased fatty acid synthetase, stearoyl-CoA desaturase, acyl-CoA synthetase long-chain family member 1 and acetyl-CoA carboxylase expression to inhibit NEFA synthesis. In addition, the CHM treatment increased plasma insulin (INS) levels and up-regulated insulin receptors (INSR) glycerol-3-phosphate acyltransferase and mitochondrial and 1-acylglycerol-3-phosphate O-acyltransferase 6 mRNA expression to enhance TG synthesis. The protein levels of phosphatidylinositol 3-hydroxy kinase (PI3K), serine/threonine protein kinase (Akt), S6 kinase (S6K), phospho-serine/threonine protein kinase (p-Akt), and phospho-S6 kinase (p-S6K) also were increased. PI3K, p-Akt, and p-S6K were significantly up-regulated in the liver. These data suggest that the CHM may be a beneficial compound to decrease plasma NEFA and TG levels by activating the INS/INSR-PI3K-Akt-SREBP1c pathway resulting in fat content reduction in the carcass.

Chinese herbal product (CHP) is the major type of traditional Chinese medicine (TCM) and widely used to relief the symptom of colorectal cancer. The aim of the study was to analyze the utilization of CHP for treating patients with colorectal cancer in Taiwan. The usage of CHP, frequency of services, and prescription pattern for colorectal cancer were analyzed from a randomly sampled cohort of 1 million beneficiaries from the National Health Insurance Research Database. The odds ratios for utilization of CHP were estimated with logistic regression model. 2846 patients were newly diagnosed as colorectal cancer during 1998-2008 in the million cohort in Taiwan. 42.7% (n = 1214) of them used CHP. Colorectal cancer was the most common diagnosis coded by TCM doctor, followed by symptoms, signs, and ill-defined conditions. Costusroot and Amomum Six Gentlemen Decoction ( xiāng shā liù jūn zǐ tāng) was the most frequently prescribed formula for treating colorectal cancer. Among the top 10 most frequently prescribed CHP for treating colorectal cancer, six containing Ginseng Radix (, ​Panax ginseng) and two containing Astragali Radix (, Astragalus membranaceus), which are reported to have potential beneficial synergistic effects on colorectal cancer cells. CHP containing Ginseng Radix or Astragali Radix are the most frequently prescribed for colorectal cancer and their effects should be taken into account by healthcare providers.

Ethnopharmacological relevance: Rheum palmatum L; Astragalus membranaceus (Fisch.), is referred to as 'Dahuang, Huangqi' in China. As an important medicinal plant, the rhizome of rhubarb and astragalus is traditionally used in the treatment of kidney diseases associated with renal failure, inflammation and tumors.

Aim of the study: This study aimed to investigate the effect of a drug-containing serum of rhubarb-astragalus capsules (composed of rhubarb and astragalus) and to elucidate its mechanism in the epithelial-mesenchymal transformation of renal tubular epithelial cells.

Materials and methods: Epithelial-mesenchymal transformation (EMT) of HK-2 cells was induced by TGF-β1, and rhubarb-astragalus and losartan drug-containing serum from rats, as well as SB203580 (a specific inhibitor of p38 MAPK), were used. High-performance liquid chromatography analysis was performed to determine the main components of the drug-containing serum of rhubarb-astragalus from rats. Western blotting and immunofluorescence analysis were used to determine the levels of protein expression, and real-time quantitative PCR analysis was used to detect the levels of gene expression.

Results: The drug-containing serum of rhubarb-astragalus contained emodin (0.36 μg/ml) and danthraquinone (0.96 μg/ml). Rhubarb-astragalus significantly decreased the protein expression levels of α-SMA, FN, vimentin and N-cadherin in HK-2 cells that were increased by TGF-β1, while it significantly increased the E-cadherin protein expression level that was decreased by TGF-β1. Rhubarb-astragalus also significantly decreased the protein expression levels of TGF-β1 and p38 MAPK and the mRNA expression levels of α-SMA, vimentin, TGF-β1, p38 MAPK, Smad2 and Smad3 in HK-2 cells that were increased by TGF-β1. It is worth noting that SB203580 (a p38 MAPK inhibitor) had similar effects as rhubarb-astragalus in this study.

Conclusion: The drug-containing serum of rhubarb-astragalus can inhibit EMT in HK-2 cells by downregulating the TGF-β1/p38 MAPK/Smad2/3 pathway.

Keywords: Drug-containing serum; Epithelial-mesenchymal transformation; Renal tubule epithelial cells; Rhubarb-astragalus; TGF-β1/p38 MAPK/Smad.

Metabolomics, an important part of systems biology, can reveal the complex pathogenesis of many diseases and mechanism of Chinese materia medica (CMM). Astragalus membranaceus-Curcuma wenyujin (AC) was a classic drug pair that has a good clinical effect on gastrointestinal inflammation and many tumors. Our previous research proved that AC can inhibit tumor growth and metastasis especially the colorectal cancer (CRC), also promote the normalization of tumor blood vessels, but its optimal ratio and the specific mechanism is still not clear. In this study, colon cancer mice of orthotopic transplantion model was used to screen the best proportion, UPLC-Q-TOF/MS metabolomics analysis method was established to explore the pathogenesis of colon cancer and the molecular mechanism of AC. The correlation analysis of metabolite changes and tumor growth was analyzed by R language. The result showed that AC at the ratio of 2:1 showed the best effect on inhibiting tumor growth, also the liver and spleen metastasis rate. A total of 23 potential biomarkers were detected in the serum of colon cancer mice by the analysis of Progenesis QI (Version 2.4) software. Among this, 11 metabolites including purines, steroids, phytosphingosine and l-palmitoylcarnitine were up-regulated in CC mice, while 12 metabolites like amino acids, deoxyribose and dihydrobiopterin were down-regulated in CC mice. After the treatment of AC for 15 days, 8 biomarkers were up-regulated, and 9 biomarkers down-regulated. Especially, AC at the ratio of 2:1 showed a significant callback effect on metabolic biomarkers, such as hypoxanthine, xanthosine, 7-methylxanthine, all-trans-retinoic acid, dihomo-γ-linolenic acid. 8 metabolic pathways: Aminoacyl-tRNA biosynthesis, Nicotinate and nicotinamide metabolism, Phenylalanine, tyrosine and tryptophan biosynthesis, Valine, leucine and isoleucine biosynthesis, Phenylalanine metabolism, Caffeine metabolism, Retinol metabolism, Alanine, aspartate and glutamate metabolism were selected as the model group disturbed metabolic pathways after the enrichment of MetaboAnalyst 4.0 online analysis software. And compared with the model group, Valine, leucine and isoleucine biosynthesis, Aminoacyl-tRNA biosynthesis, Caffeine metabolism pathway and Retinol metabolism pathways were altered after the intervention of AC. The correlation analysis results showed that various endogenous metabolites in serum have a strong correlation with tumor weight, such as hypoxanthine, which provides a basis for the selection of clinical markers. The results showed that AC can partially regulate metabolic disorder of CC mice by reversing the changes of metabolites, so as to inhibit the growth and metastasis of colon cancer, especially at the ratio of 2:1. These findings can provide a scientific basis for exploring the diagnostic biomarkers of colon cancer, and for clinical application of AC in the treatment of CRC program.

Keywords: Astragalus membranaceus-Curcuma wenyujin; Biomarkers; Colorectal cancer; Metabolic pathway; Metabolomics.

Astragalus Polysaccharide (APS) is the main component of Astragalus membranaceus, an anti-diabetic herb being used for thousands of years in Chinese Traditional Medicine (TCM). In this study, we aimed to evaluate the impact of APS on hepatic insulin signaling, autophagy and ER stress response in high-fat-diet (HFD)-induced insulin resistance (IR) mice. APS was intra gastrically administrated and metformin was used as a control medicine. Apart from monitoring the changes in the important parameters of IR progression, the gene and protein expression of the key factors marking the state of hepatic ER stress and autophagic flux was examined. We found that, largely comparable to the metformin regime, APS treatment resulted in an overall improvement of IR, as indicated by better control of body weight and blood glucose/lipid levels, recovery of liver functions, and regained insulin sensitivity. In particular, the excessive and pro-apoptotic ER stress response and inhibition of autophagy, as a result of prolonged HFD exposure, were significantly corrected by APS administration, indicating a switch of the cellular fate in favor of cell survival. Using the HepG2/IR cell model, we demonstrated that APS modulated the insulin-initiated phosphorylation cascades in a similar manner as metformin. This study provides a rationale for exploiting the insulin-sensitizing potential of APS, which has a therapeutic performance almost equivalent to metformin, to enrich our options in the treatment of IR.

Chitosan, behaving as a potent biotic elicitor, can induce plant defense response with the consequent enhancement in phytoalexin accumulation. Accordingly, chitosan elicitation was conducted to promote the production of two phytoalexins, i.e. formononetin and calycosin (also known as health-promoting isoflavones), in Astragalus membranaceus hairy root cultures (AMHRCs). Compared with control, 12.45- and 6.17-fold increases in the yields of formononetin (764.19 ± 50.81 μg/g DW) and calycosin (611.53 ± 42.22 μg/g DW) were obtained in 34 day-old AMHRCs treated by 100 mg/L of chitosan for 24 h, respectively. Moreover, chitosan elicitation could cause oxidative burst that would induce the expression of genes (MPK3 and MPK6) related to mitogen-activated protein kinase signaling (MAPK) cascades, which contributed to the transcriptional activation of pathogenesis-related genes (β-1,3-glucanase, Chitinase, and PR-1) and eight biosynthesis genes involved in the calycosin and formononetin pathway. Overall, the findings in this work not only highlight a feasible chitosan elicitation practice to enhance the in vitro production of two bioactive isoflavones for nutraceutical and food applications, but also contribute to understanding the phytoalexin biosynthesis in response to chitosan elicitation.

The purpose of this study was to investigate the effect of Astragalus membranaceus (AM) on erythrocyte sodium content and sodium transport in the coronary heart disease(CHD). 67 cases of CHD was randomly divided into treatment group (TG) and control group (CG), TG (45 cases) were treated with AM intravenous 24 g/d infused into 5% glucose 250 ml, CG (22 cases) with 5% glucose 250 ml. After a curable stage of 14 days, results showed that the erythrocyte sodium content was markedly decreased and the sodium pump activity was significantly increased (P less than 0.01 respectively) in TG; but no significant difference in membrane permeability to sodium was observed. There were no difference in erythrocyte sodium content and sodium transport in CG. It was suggested that AM had the effects to increase erythrocyte activity of sodium pump.

Astragaloside IV (AsIV), one of the major active ingredients in Astragalus membranaceus, has demonstrated remarkable antifibrotic effects via its antioxidative activity. Cardiac fibrosis is an important pathological mechanism during cardiac remodelling associated with heart failure. In the present study, the mechanism underlying the antifibrotic effect of AsIV upon isoprenaline (ISO) stimulation was investigated. AsIV significantly improved cardiac fibrosis in vivo and dose‑dependently inhibited ISO‑induced CF proliferation in vitro. The ISO‑triggered elevation in reactive oxygen species (ROS) levels was remarkably inhibited by AsIV, as well as ROS scavenger N‑acetylcysteine (NAC), and not affected by cardiotrophin‑1 (CT‑1) knockdown. In addition, AsIV effectively reversed ISO‑induced upregulation of CT‑1 expression, which was blunted by pretreatment with NAC. Cardiac fibroblast (CF) proliferation and collagen Ι overexpression induced by ISO stimulation were effectively abrogated by AsIV, NAC, and CT‑1 small interfering RNA transfection. Taken together, these results demonstrated that AsIV was able to effectively inhibit ISO‑induced CF proliferation and collagen production through negative regulation of ROS‑mediated CT‑1 upregulation.

Astragalus polysaccharide (APS) is known to have a variety of pharmacological activities. In the present study, enzyme assisted extraction of APS from Astragalus mongholicus using various enzymes were examined. Research found that glucose oxidase offered a better performance in enhancement of extraction yields of APS than other ones. Glucose oxidase assisted extraction process was further optimized by using response surface method (RSM) to obtain maximum yield of crude APS. The optimized extraction conditions were as follows: enzyme amount of 3.0%, enzyme treated time of 3.44 d, enzyme treated temperature of 56.9 °C and extraction solvent pH of 7.8. Under these conditions, the experimental yield was 29.96 ± 0.14%, which was well in close agreement with the value (30.19%) predicted by RSM model and increased more than 250% compared with none enzyme treated ones. Pharmacological test showed that enzyme assisted APS had a better antioxidant activity (about 2 times higher) than none enzyme treated ones.

The effects of Astragalus membranaceus and Lycium barbarum on the growth performance and non-specific immunity of grass carp and Nile tilapia were studied. Herb extracts of Chinese medicinal herbs (2 g kg-1 or 20 g kg-1) were incorporated into food waste based fish feed pellets. Fish growth and selected non-specific immune parameters of grass carp and Nile tilapia were studied in two separate feeding trials. Both grass carp and Nile tilapia fed diets of feed pellets containing 2 g kg-1Lycium barbarum extract achieved the best relative weight gain, feed conversion ratio, specific growth rate and protein efficiency ratio among all experimental diets. Fish fed with diets containing 2 g kg-1Lycium barbarum also resulted in significantly higher total immunoglobin, bactericidal activity and anti-protease activity; and also a lower mortality when challenged with pathogenic bacteria. On the other hand, both fish species fed with diets containing 20 g kg-1 of Astragalus membranaceus and 20 g kg-1Lycium barbarum, resulted in significantly impaired weight gain. In addition, incorporation of 2 g kg-1Lycium barbarum extract would be a more suitable dose for both fish species, in terms of achieving better feed conversion ratio, specific growth rate, protein digestibility, and improved non-specific immune parameters. Based on this study, it can be concluded that waste based feed pellets incorporated with Chinese medicinal herb extracts have the ability to enhance growth and immunity of fish. Therefore, the use of Chinese medicinal herbs in aquaculture should be encouraged, in order to replace certain antibiotics known to impose environmental and health effects through the discharge of aquaculture effluents.

Jixuepaidu Tang-1 is obtained from the decoction of the Chinese traditional medicinal plants including Centella asiatica, Astragalus membranaceus, and Sanguis draconis. Transforming growth factor-β1 (TGF-β1)/serum- and glucocorticoid-inducible kinase-1 (SGK1)-induced epithelial-mesenchymal transition (EMT) plays a pivotal role in the pathogenesis of diabetic nephropathy (DN). In addition, long non-coding RNAs (lnRNAs) participate in the development of DN, but the role of lncRNA LOC498759 in DN is still unclear. This study aims to investigate the role of Jixuepaidu Tang-1 in regulating podocyte injury and renal damage in DN and to validate whether the mechanisms involve TGF-β1/SGK1 signaling and LOC498759. The drug treatment was initiated 2 weeks after the DN modeling. The MTT method and TUNEL staining were used to measure cell viability and apoptosis, respectively. Immunofluorescence staining was used to detect the expression of nephrin and desmin in podocytes. Sera from the Jixuepaidu Tang-1-treated mice reversed the high glucose (HG)-induced podocyte injury and EMT in mouse podocytes. Further in vivo assay revealed that Jixuepaidu Tang-1 not only reduced the ratio of the kidney to body weight, 24 h-urine total protein, and blood glucose, but alleviated glomerular mesangial extracellular matrix deposition and glomerular cell apoptosis in the streptozotocin-induced DN mice. Mechanically, the mechanisms of Jixuepaidu Tang-1 may involve the suppression of EMT by inhibiting the TGF-β1/SGK1-induced LOC498759 expression. Collectively, Jixuepaidu Tang-1 attenuates podocyte injury and renal damage in DN, and inhibits EMT through suppressing TGF-β1/SGK1-LOC498759 signaling.

Spinal cord injury (SCI) is a catastrophic disease associated with damaged neurological structures and has become a significant social and economic burden for the health care system and patients' families. The use of Chinese Herbal Medicine (CHM) to treat SCI has been increasing in recent years. This meta-analysis aimed to investigate the effectiveness of CHM for patients with SCI. Therefore, we included randomized controlled trials (RCTs) of CHM for SCI in seven databases. A total of 26 studies involving 1961 participants were included in this study. No serious heterogeneity or publication bias was observed across each study. The results showed that significant improvements of the American Spinal Injury Association (ASIA)-grading improvement rate ([Formula: see text], [Formula: see text]), clinical effective rate ([Formula: see text], [Formula: see text]), ASIA motor score ([Formula: see text], [Formula: see text]), ASIA sensory score (total) ([Formula: see text], [Formula: see text]), ASIA sensory score (light touch) ([Formula: see text], [Formula: see text]), ASIA sensory score (pinprick) ([Formula: see text], [Formula: see text]), and activities of daily living (ADL) score ([Formula: see text], [Formula: see text]) in CHM group compared with the control group. Among the CHM groups, Buyang Huanwu decoction was the most frequently prescribed herbal formula, while Astragalus membranaceus was the most commonly used single herb. In addition, there were no serious and permanent adverse effects in the two groups. The methodological quality of the most included RCTs was poor and the quality of evidence for the main outcomes was from very low to moderate according to the GRADE system. Current evidence suggests that CHM is an effective and safe treatment for SCI and could be treated as a complementary and alternative option with few side effects. However, considering the low quality, small size, and high risk of the studies identified in this meta-analysis, higher methodological quality, rigorously designed RCTs with large sample sizes are needed to confirm the results.

Keywords: Chinese Herbal Medicine; Efficacy; Meta-Analysis; Review; Safety; Spinal Cord Injury.

Astragalus mongholicus (MG) and Astragalus membranaceus (MJ), both generally known as Huangqi in China, are two perennial herbals widely used in variety diseases. However, there were still some differences in the chemical ingredients between MG and MJ. In this paper, metabolomics combined with the ultra-high performance liquid chromatography coupled with electrospray ionization/quadrupole time-of-flight mass spectrometry (UHPLC-ESI-Q-TOF-MS/MS) was employed to contrastively analyze the chemical constituents between MG and MJ. As a result, principal component analysis showed that MG and MJ were separated clearly. A total of 53 chemical markers were successfully identified for the discrimination of MG and MJ. Of them, the contents of 36 components including Astragaloside I~III, Astragaloside IV, Agroastragaloside I, etc. in MJ were significantly higher than those in MG. On the contrary, the contents of 17 other components including coumaric acid, formononetin, sophoricoside, etc. in MG were obviously higher than those in MJ. The results showed that the distinctive constituents in MG and MJ were remarkable, and MJ may own stronger pharmacological activities than MG. In a word, MG and MJ may be treated as two different herbs. This paper demonstrated that metabolomics was a vitally credible technology to rapidly screen the characteristic chemical composition of traditional Chinese medicine.

Astragalus membranaceus (A. membranaceus) is commonly used in various herbal formulations to treat several human and animal diseases. Polysaccharides, which are the major bioactive components in the A. membranaceus, exhibit various bioactive properties. However, the ability of A. membranaceus polysaccharides (APS) to activate the mucosal immune response has not been examined. We examined the effect of intranasal administration of APS on mucosal immune cell activation and the growth-inhibitory activity against pulmonary metastatic melanoma in mice by combination treatment with immune checkpoint blockade. The intranasal treatment of APS increased the number of lineage^-CD11c⁺ dendritic cell (DCs) in the mesenteric lymph nodes (mLN) through the upregulation of CC-chemokine receptor 7 expression. Moreover, intranasal treatment of APS activated DCs, which further stimulated natural killer (NK) and T cells in the mLN. The APS/anti-PD-L1 antibody combination inhibited the pulmonary infiltration of B16 melanoma cells. The depletion of NK cells and CD8 T cells in mice mitigated the anti-cancer effect of this combination, thereby highlighting the critical role of NK cells and CD8 T cells in mediating anti-cancer immunity. These findings demonstrated that APS could be used as a topical mucosal adjuvant to enhance the immune check point inhibitor anti-cancer effect.

Keywords: Anti-cancer; Astragalus membranaceus polysaccharides; Cytotoxic T lymphocyte; Immune checkpoint inhibitor; Mucosal immunity; Natural killer cell.

Ethnopharmacological relevance: Qizhen capsule (QZC) is a traditional Chinese medicine (TCM) preparation that has been widely used in clinical practice and exerts promising therapeutic effects against breast, lung, and gastric cancers. However, studies have not reported whether QZC inhibits colorectal cancer (CRC) development and progression. Meanwhile, the underlying molecular mechanisms of its anticancer activity have not been studied.

Aim of the study: To investigate the anticancer effects of QZC on CRC and the possible underlying molecular mechanisms of QZC in vitro and in vivo.

Materials and methods: The MTT assay and flow cytometry were used to determine the viability and apoptosis of HCT116 and HT-29 cancer cells. A xenograft nude mouse model was used to study the antitumor effects of QZC in vivo. Western blotting was performed to determine the expression of key proteins responsible for the molecular mechanisms elicited by QZC. Immunofluorescence staining was performed to detect the expression of nonsteroidal anti-inflammatory drug (NSAID)-activated gene-1 or growth differentiation factor-15 (NAG-1/GDF15). Small interfering RNAs (siRNAs) were used to silence NAG-1/GDF15 in cells.

Results: In this study, QZC significantly reduced the viability of HCT116 and HT-29 cells and induced apoptosis in dose- and time-dependent manners, but displayed much less toxicity toward normal cells. QZC-induced apoptosis in HCT116 cells was accompanied by the deregulation of the expression of the Bcl-2, Bax, PARP, caspase-3, and caspase-9 proteins. Furthermore, QZC induced NAG-1/GDF15 expression in HCT116 cells, while silencing of NAG-1/GDF15 attenuated QZC-induced apoptosis and cell death. Next, QZC increased the phosphorylation of mTOR, AMPK, p38, and MAPK/ERK in HCT116 cells. We then demonstrated that QZC-induced apoptosis and NAG-1/GDF15 upregulation were mediated by MAPK/ERK activation. Moreover, QZC significantly inhibited HCT116 xenograft tumor growth in nude mice, which was accompanied by NAG/GDF15 upregulation and MAPK/ERK activation. QZC also prevented 5-FU-induced weight loss or cachexia in tumor-bearing mice. The expression of Ki67 and PCNA was suppressed, while cleaved caspase-3 level and TUNEL staining were increased in the tumor sections from QZC-treated mice compared to the control.

Conclusion: QZC is a novel anticancer agent for CRC that targets NAG-1/GDF15 via the MAPK/ERK signaling pathway.

Keywords: Astragalus membranaceus; Colorectal cancer; MAPK/ERK; NAG-1/GDF15; Panax notoginseng; apoptosis.

Osteoarthritis (OA) is the common form of arthritis and is characterized by disability and cartilage degradation. Although natural product extracts have been reported to have anti-osteoarthritic effects, the potential bioactivity of Ryupunghwan (RPH), a traditional Korean medicinal botanical formula that contains Astragalus membranaceus, Turnera diffusa, Achyranthes bidentata, Angelica gigas, Eclipta prostrata, Eucommia ulmoides, and Ilex paraguariensis, is not known well. Therefore, the inhibitory effects of single compounds isolated from RPH on the OA-related molecules were investigated using IL-1β-stimulated chondrosarcoma SW1353 (SW1353) cell model. Two bioactive compounds, isomucronulatol 7-O-β-d-glucoside (IMG) and ecliptasaponin A (ES) were isolated and purified from RPH using column chromatography, and then the structures were analyzed using ESI-MS, ¹H-NMR, and 13C-NMR spectrum. The expression or amount of matrix metalloproteinase 13 (MMP13), COX1/2, TNF-α, IL-1β or p65 was determined by RT-PCR, Western blot, and enzyme-linked immunosorbent assay (ELISA). RPH pretreatment reduced the expression and amounts of MMP13, and the expression of collagen II, COX1/2, TNF-α, IL-1β or p65, which were increased in IL-1β-stimulated SW1353 cells. IMG reduced the expression of all OA-related molecules, but the observed inhibitory effect was less than that of RPH extract. The other single compound ES showed the reduced expression of all OA-related molecules, and the effect was stronger than that in IMG (approximately 100 fold). Combination pretreatment of both single components remarkably reduced the expression of MMP13, compared to each single component. These synergic effects may provide potential molecular modes of action for the anti-osteoarthritic effects of RPH observed in clinical and animal studies.

The dried root of Astragalus membranaceus is a well-known herbal medicine, and it is useful in treating chronic diseases and weakness, as well as for improving overall health and vitality. Astragalosides, which are root quality indicators of A. membranaceus, are natural triterpenoid saponins that are used in the treatment of diabetes and cardiovascular diseases. Currently, there is an urgent need to improve their production because of their low quantity in plants and the difficulty of chemical synthesis. In this study, yeast extract was added to facilitate elicitation in Agrobacterium-mediated hairy root cultures, thereby enhancing astragaloside production in A. membranaceus. Results showed that yeast extract could stimulate astragaloside content effectively in the hairy roots of A. membranaceus. Moreover, astragaloside accumulation was positively correlated with the upregulation of mevalonate biosynthetic gene expression in the presence of yeast extract. Our study demonstrated that pretreatment with yeast extract (3.65 mM) for 72 h serves as an effective strategy to enhance astragaloside levels in A. membranaceus hairy root cultures. Thus, these optimal conditions can provide valuable information for the improvement of astragaloside industrial production in A. membranaceus.

Keywords: Astragalus membranaceus; Astragaloside; elicitation; hairy root culture; yeast extract.

Ageing is an unavoidable, universal, biological phenomenon affecting all organisms, which involves variable declines of individuals motor and memory capabilities. This study aimed to investigate the potential ameliorating effects of curcumin C3 complex, Astragalus membranaceus and blueberry on certain age-related biochemical alterations in rat liver. Four groups of rats, aged 12 months-old, were used. The first group; aged control group in which rats were left without any treatment until the age of 17 months. The other three groups received daily by oral gavage for 5 months the following supplements; curcumin C3 complex (110 mg/kg), Astragalus membranaceus (100 mg/kg) and blueberry (100 mg/kg) respectively. Additionally, a fifth group of rats, aged 5 months-old, was used as an adult control group. Our supplements alleviated ageing-induced redox state imbalance and inflammation as evidenced by reduction of hepatic thiobarbituric acid reactive substances and 8-hydroxydeoxyguanosine levels, restoration of total antioxidant capacity and nitric oxide contents, and lessening of lipofuscin deposition. All supplements decreased hepatic interlukin-6 gene expression and serum levels. Notably, Astragalus membranaceus and blueberry upregulated hepatic telomerase reverse transcriptase gene expression and increased telomere length. Our findings recommend the use of these natural hepatoprotective supplements for the elderly to promote healthy ageing and minimize the risk of age-related liver diseases.

Keywords: Ageing; Astragalus membranaceus; Blueberry; Curcumin C3 complex; Liver; Telomeres.

Nature, a vast reservoir of pharmacologically active molecules, has been most promising source of drug leads for the cure of various pathological conditions. Formononetin is one of the bioactive isoflavones isolated from different plants mainly from Trifolium pratense, Glycine max, Sophora flavescens, Pycnanthus angolensis, and Astragalus membranaceus. Formononetin has been well-documented for its anti-inflammatory, anticancer, and antioxidant properties. Recently anticancer activity of formononetin is widely studied. This review aims to highlight the pharmacological potential of formononetin, thus providing an insight of its status in cancer therapeutics. Formononetin fights progression of cancer via inducing apoptosis, arresting cell cycle, and halting metastasis via targeting various pathways which are generally modulated in several cancers. Although reported data acclaims various biological properties of formononetin, further experimentation on mechanism of its action, medicinal chemistry studies, and preclinical investigations are surely needed to figure out full array of its pharmacological and biological potential.