LPS-induced macrophage exosomes promote the activation of hepatic stellate cells and the intervention study of total astragalus saponins combined with glycyrrhizic acid.

Huangqi decoction, also known as Huangqi Liuyi decoction, was first recorded in the prescriptions of the Bureau of Taiping People's Welfare Pharmacy. It comprises astragalus and licorice, which is a commonly used prescription in traditional Chinese medicine for the clinical treatment of chronic liver disease, especially liver cirrhosis. Total astragalus saponins (AST) is the main component of astragalus, and glycyrrhizic acid (GA) is the main component of licorice. In this study, normal macrophage exosomes were extracted, and the exosomes incubated with lipopolysaccharides (LPS) and those incubated with LPS + AST + GA were co-cultured with JS1 cells (hepatic stellate cell line). The survival rate and the activation of key signaling pathways of JS1 cells in each group were detected and compared. We found that the co-culture of LPS-induced macrophage exosomes with JS1 cells could significantly increase the expression levels of Collagen-1 (Col-1) and Alpha smooth muscle actin (α-SMA)in JS1 cells. However, a significant reversal effect was observed after pretreatment with AST combined with GA. Further evaluation found that the expression levels of phospho (p)-Smad2 and p-Smad3 in the JS1 cells were significantly increased after macrophages were induced with LPS, whereas pretreatment with AST + GA could significantly decrease the expression levels of p-Smad2 and p-Smad3. Preliminary results of this study indicated that LPS-induced macrophage exosomes can promote the activation of hepatic stellate cells, and the pretreatment of AST combined with GA can exert a significant intervention effect. In this study, the new mechanism of anti-hepatic fibrosis effect of traditional Chinese medicine components of Huangqi Decoction was analyzed from the perspective of exosomes.

Glycyrrhizic Acid against Mycoplasma gallisepticum-Induced Inflammation and Apoptosis Through Suppressing the MAPK Pathway in Chickens.

Mycoplasma gallisepticum (MG) is the primary pathogen of chronic respiratory diseases (CRDs) in chickens. In poultry production, antibiotics are mostly used to prevent and control MG infection, but the drug resistance and residue problems caused by them cannot be ignored. Glycyrrhizic acid (GA) is derived from licorice, a herb traditionally used to treat various respiratory diseases. Our study results showed that GA significantly inhibited the mRNA and protein expression of pMGA1.2 and GapA in vitro and in vivo. Furthermore, the network pharmacology study revealed that GA most probably resisted MG infection through the MAPK signaling pathway. Our results demonstrated that GA inhibited MG-induced expression of MMP2/MMP9 and inflammatory factors through the p38 and JUN signaling pathways, but not the ERK pathway in vitro. Besides, histopathological sections showed that GA treatment obviously attenuated tracheal and lung damage caused by MG invasion. In conclusion, GA can inhibit MG-triggered inflammation and apoptosis by suppressing the expression of MMP2/MMP9 through the JNK and p38 pathways and inhibit the expression of virulence genes to resist MG. Our results suggest that GA might serve as one of the antibiotic alternatives to prevent MG infection.

Glycyrrhizic Acid Alleviates Lipopolysaccharide (LPS)-Induced Acute Lung Injury by Regulating Angiotensin-Converting Enzyme-2 (ACE2) and Caveolin-1 Signaling Pathway.

Acute lung injury (ALI) is mainly caused by severe infection, shock, trauma, and burn, which causes the extensive release of inflammatory factors and other mediators. As a major bioactive constituent of traditional Chinese herb licorice, glycyrrhizic acid (GA) plays an important effect on inflammatory regulation. Nevertheless, the exact mechanism of this effect remains unclear. The present study aims to explore the potential protective effect of GA on LPS-induced ALI. Our results showed that GA significantly attenuated LPS-induced ALI and decreased the production of inflammatory factors, including IL-1ß, MCP-1, COX2, HMGB1, and adhesion molecules, such as E-selectin, VCAM-1, and modulated expression of angiotensin-converting enzyme 2 (ACE2). Moreover, treatment of ACE2 inhibitor (MLN-4760) reversed the effects of GA on the secretion of pro-inflammatory factors in ALI. Additionally, GA exerts its protective effect by regulating the ACE2 and caveolin-1/NF-κB signaling pathway. In conclusion, this study showed that GA alleviated LPS-induced ALI by upregulating ACE2 and inhibiting the caveolin-1/NF-κB signaling pathway.

Study on antioxidant potential, immunological response, and inflammatory cytokines induction of glycyrrhizic acid (GA) in silver carp against vibriosis.

Effect of dietary with 100, 200, and 300 mg kg-1 glycyrrhizic acid (GA) on growth enhancer, blood physiology, digestive-antioxidant enzyme ability, innate-adaptive defense, and inflammatory cytokines induction was studied in silver carp, Hypophthalmichthys molitrix against vibriosis caused by Vibrio alginolyticus. Significant weight gain (WG), specific growth rate (SGR), and 100% survival rate (SR) was attained non-infected health (NiH) fish fed in control or all GA diets on 30, 45, and 60 days. Both NiH and V. alginolyticus challenged (VaC) fish treated with 200 mg GA diet significantly (P < 0.05) exhibited an enhancement in leucocytes value on 30, 45, and 60 days. Albumin (AB) or total proteins (TP) levels were significantly (P < 0.05) better in both groups fed 200 GA on 45 and 60 days. Malondialdehyde (MDA) and superoxide dismutase (SOD) activities were also substantial (P < 0.05) in both groups fed 200 mg GA on days 30, 45, and 60; whereas glutathione peroxidase (GPx) and catalase (CAT) activities were significantly (P < 0.05) better in both groups received 200 mg GA on days 45 and 60. Phagocytic (PC) and lysozyme (Lyz) activities significantly enhanced in both groups fed 200 or 300 mg GA on 45 and 60 days. Respiratory burst (RB), reactive oxygen species (ROS) and immunoglobulin (Ig) production significantly (P < 0.05) increased in both groups administered 200 or 300 mg GA. Growth hormone (GH) mRNA was up regulated in 200 mg GA trial on 45 days and in 200 or 300 mg GA treatments on 60 days. The IL-8 cytokine mRNA expression was up-regulated in both groups 200 and 300 mg GA on days 45 and 60, whereas TNF-α mRNA expression was increased in 200 mg GA. In addition, IL-10 cytokine mRNA expression was up regulated in 200 mg GA on 45 days whereas it was increased in both 200 mg and 300 mg GA trial on 60 days. The present study revealed that feeding fish 200 mg GA per kg diet demonstrated a better growth, digestive-antioxidant activity, innate-adaptive defense, and inflammatory cytokines induction than lower or higher dosage of GA in H. molitrix against V. alginolyticus.

Bacillus amyloliquefaciens Rescues Glycyrrhizic Acid Loss Under Drought Stress in Glycyrrhiza uralensis by Activating the Jasmonic Acid Pathway.

Drought is a major factor limiting the production of the perennial medicinal plant Glycyrrhiza uralensis Fisch. (Fabaceae) in Northwest China. In this study, 1-year-old potted plants were inoculated with the strain Bacillus amyloliquefaciens FZB42, using a gradient of concentrations (CFU), to test for microbe-induced host tolerance to drought condition treatments in a greenhouse experiment. At the concentration of 108 CFU ml-1, FZB42 had significant growth-promoting effect on G. uralensis: the root biomass was 1.52, 0.84, 0.94, and 0.38 times that under normal watering and mild, moderate, and severe drought stress conditions, respectively. Under moderate drought, the positive impact of FZB42 on G. uralensis growth was most pronounced, with both developing axial and lateral roots strongly associated with indoleacetic acid (IAA) accumulation. An untargeted metabolomic analysis and physiological measurements of mature roots revealed that FZB42 improved the antioxidant system of G. uralensis through the accumulation of proline and sucrose, two osmotic adjustment solutes, and by promoting catalase (CAT) activity under moderate drought stress. Furthermore, significantly higher levels of total flavonoids, liquiritin, and glycyrrhizic acid (GA), the pharmacologically active substances of G. uralensis, were found in the roots of inoculated plants after FZB42 inoculation under all imposed drought conditions. The jasmonic acid (JA) content, which is closely related to plant defense responses and secondary metabolites' production, was greatly increased in roots after the bacterial inoculations, indicating that FZB42 activated the JA pathway. Taken together, our results demonstrate that inoculation with FZB42 alleviates the losses in production and pharmacological metabolites of G. uralensis caused by drought via the JA pathway's activation. These results provide a developed prospect of a microbial agent to improve the yield and quality of medical plants in arid and semi-arid regions.

Effects of glycyrrhizin on lipopolysaccharide-induced acute lung injury in a mouse model.

BACKGROUND: Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are serious clinical disease entities characterized by inflammatory pulmonary edema, which lead to acute hypoxic respiratory failure through various etiologies. According to the studies to date, ALI/ARDS has been recognized as a form of multiorgan failure related to overactive immune response, and overproduction of proinflammatory cytokines released from activated inflammatory cells are considered to play a key role in the development of ALI. Glycyrrhizin (GL) is an extractive component derived from Glycyrrhiza glabra (licorice), which has recently been reported to have various pharmacological effects like anti-inflammatory, anti-tumor, hepato-protective, and anti-viral activities. Nevertheless, the therapeutic effect of GL in ALI is still unclear. The aim of this study was to investigate therapeutic effects of GL on lipopolysaccharide (LPS)-induced ALI in a mouse model and to elucidate explicable mechanisms involved. METHODS: A total of 36 BALB/c mice (6-week-old, 27.7±1.9-gram body weight) were randomly divided into 3 groups: the control group (normal saline was administered intravenously, n=10), the LPS group (LPS 50 mg/kg was intraperitoneally administered, n=13), and the LPS + GL group (GL was administered intravenously immediately and 12 hours after LPS injection, n=13). Mice were sacrificed after 24 hours, and bronchoalveolar lavage fluid (BALF) was collected for the estimation of protein content, inflammatory cell counts, proinflammatory cytokines, myeloperoxidase (MPO) activity, and expressions of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and nuclear factor kappa B (NF-κB). Then, the lungs were excised for molecular target, histopathological, and immunohistochemical examinations. RESULTS: Compared to the LPS group, GL significantly decreased protein content, inflammatory cell counts, tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α), IL-6, MPO activity, and expressions of COX-2, iNOS, and NF-κB in the LPS + GL group. GL attenuated migration and infiltration of inflammatory cells, showing a marked decrease in CD 11b-positive cells (26.77%±0.83% vs. 41.77%±0.81% vs. 23.23%±1.92%, P<0.05) as well as CXCR4-/CXCR1-positive cells (CXCR4: 37.23%±1.00% vs. 59.37%±2.37% vs. 47.45%±4.36%; CXCR1: 32.10%±1.56% vs. 47.03%±1.99% vs. 21.70%±6.50%; all P<0.05) in the control, LPS, and LPS + GL groups. Additionally, immunohistochemistry showed that the expression of Toll-like receptor 4 (TLR-4) was inhibited by GL. CONCLUSIONS: The results of this study indicate that GL may have anti-inflammatory and protective effects on LPS-induced ALI in mice. GL inhibited proinflammatory cytokines playing a key role in the initial phase of inflammatory response, which suggests that inhibition of the TLR-4/NF-κB signal pathway would be a possible mechanism underlying the action of GL. Thus, GL can be used as a novel therapeutic strategy for pulmonary inflammation.

Glycyrrhizic acid ameliorates LPS-induced acute lung injury by regulating autophagy through the PI3K/AKT/mTOR pathway.

Acute lung injury (ALI) is a major pathological issue characterized by serious inflammatory response, and a major clinically critical illness with high morbidity and mortality. Glycyrrhizic acid (GA) is a major bioactive constituent isolated from traditional Chinese herb licorice, which has been reported to have positive effects on inflammation. Nevertheless, the effects of GA on lipopolysaccharide (LPS)-treated ALI in mice have not been reported. The purpose of our study is to investigate the inhibitory effects of GA on ALI treated by LPS and to elucidate its possible mechanisms. We found that GA significantly attenuated lung injury and decreased the production of inflammatory factors TNF-α, IL-1ß, and HMGB1 with LPS treatment. GA induced autophagy which was showed by enhanced number of autophagosomes through upregulating the protein levels of LC3-II/I and Beclin-1 and downregulating SQSTM1/P62. Moreover, pre-treatment of 3-Methyladenine (3-MA), an autophagy inhibitor, reversed the inhibiting effects of GA on the secretion of inflammatory factors in ALI. The PI3K/AKT/mTOR pathway was associated with GA-induced autophagy under ALI induced by LPS. In conclusion, this study indicated that GA inhibited the production of inflammatory factors in LPS-induced ALI by regulating the PI3K/AKT/mTOR pathway related autophagy, which may provide a novel therapeutic perspective of GA in ameliorating ALI.

Synergistic anti-liver fibrosis actions of total astragalus saponins and glycyrrhizic acid via TGF-ß1/Smads signaling pathway modulation.

ETHNOPHARMACOLOGICAL RELEVANCE: Huangqi decoction (HQD) is a well-known traditional Chinese herbal formulation, It is an effective treatment for consumptive disease and chronic liver diseases. It consists of Radix Astragali (Astragalus membranceus(Fisch.) Bge. Root, Huangqi) and Radix Glycyrrhizae (Glycyrrhiza uralensis Fisch., root and rhizome, Gancao). Total astragalus saponins (AST) is a main component of Radix Astragali and glycyrrhizic acid(GA) is a main component of Radix Glycyrrhizae. Our primary results showed that the combination of AST and GA had an obvious synergistic effect in reducing liver collagen deposition and decreasing serum alanine aminotransferase (ALT) activity in dimethylnitrosamine (DMN)-induced liver fibrosis. AIM OF THE STUDY: Through in vivo and in vitro experiments, we aimed at investigating the key anti-fibrosis signal pathway TGF-ß1/Smads to further explore the synergistic mechanism of AST and GA. MATERIAL AND METHODS: Two hepatic fibrosis animal models, bile duct ligation-induced (BDL) and DMN-induced, were utilized. Rats were treated orally with AST, GA or AST/GA, with the effects evaluated via liver histopathology, hydroxyproline (Hyp) levels, and α-SMA expression. In the hepatic stellate cell line JS-1, cells were treated with AST/GA for 24h, followed by a cell viability assessment using Cell Counting Kit-8(CCK-8) and Real-time PCR and Western blot analysis of α-SMA, ColⅠ and TGF-ß1/Smads signaling pathway related components. RESULTS: The AST/GA combination attenuated liver tissue inflammation, collagen deposition, Hyp levels, and α-SMA expression in both BDL-and DMN-stimulated hepatic fibrosis rats. In vitro results showed that the AST/GA combination significantly inhibited JS-1 cell viability, significantly suppressed α-SMA, ColⅠ, TGF-ß1, Smad2 and Smad3 mRNA and protein expression, as well reduced p-Smad2/3. Compared with AST or GA treatment alone, the AST/GA combination significantly reduced Smad3 mRNA expression levels and TGF-ß1, Smad3, and p-Smad2/3 protein levels. CONCLUSIONS: AST and GA synergistically alleviated both BDL-and DMN-induced hepatic fibrosis via TGF-ß1/Smads signaling pathway inhibition in hepatic stellate cells.

Studies on effect of main active principles in ZHIGANCAO DECOCTION on myocardial electrophysiology / 中草药

Object To study the influence of main active principles of ZHIGANCAO DECOCTIDN (ZD), glycyrrhizic acid (GA), ginseng total saponin (GTS) and Ophiopogon total saponin (OTS) on electrophysiology of isolated rat myocardium Methods The influence of GA, GTS and OTS on the automaticity, excitability and functional refractory period of isolated rat atrium and papillary muscle were studied in comparison with ZD made free of the above said active ingredients Results The combine use of GA, GTS and OTS significantly decreased the automaticity, inhibited excitaibility and prolonged the functional refractory period of isolated rat atrium; decreased the automaticity and arrhythmia of papillary muscle induced by epinephrine, while ZD made free of GS, GTS and OTS showed much less effect than the intact decoction Conclusion GA, GTS and OTS proved to be the main effective ingredients responsible for the antiarrhythmic activity of ZD