Exercise-induced modulation of miR-149-5p and MMP9 in LPS-triggered diabetic myoblast ER stress: licorice glycoside E as a potential therapeutic target

Background:This study explores the relationship between endoplasmic reticulum(ER)stress and diabetes,particularly focusing on the impact of physical exercise on ER stress mechanisms and identifying potential therapeutic drugs and targets for diabetes-related sepsis.The research also incorporates traditional physical therapy perspectives,emphasizing the genomic insights gained from exercise therapy in disease management and prevention.Methods:Gene analysis was conducted on the GSE168796 and GSE94717 datasets to identify ER stress-related genes.Gene interactions and immune cell correlations were mapped using GeneCard and STRING databases.A screening of 2,456 compounds from the TCMSP database was performed to identify potential therapeutic agents,with a focus on their docking potential.Techniques such as luciferase reporter gene assay and RNA interference were used to examine the interactions between microRNA-149-5p and MMP9.Results:The study identified 2,006 differentially expressed genes and 616 miRNAs.Key genes like MMP9,TNF-α,and IL1B were linked to an immunosuppressive state.Licorice glycoside E demonstrated high affinity for MMP9,suggesting its potential effectiveness in treating diabetes.The constructed miRNA network highlighted the regulatory roles of MMP9,IL1B,IFNG,and TNF-α.Experimental evidence confirmed the binding of microRNA-149-5p to MMP9,impacting apoptosis in diabetic cells.Conclusion:The findings highlight the regulatory role of microRNA-149-5p in managing MMP9,a crucial gene in diabetes pathophysiology.Licorice glycoside E emerges as a promising treatment option for diabetes,especially targeting MMP9 affected by ER stress.The study also underscores the significance of physical exercise in modulating ER stress pathways in diabetes management,bridging traditional physical therapy and modern scientific understanding.Our study has limitations.It focuses on the microRNA-149-5p-MMP9 network in sepsis,using cell-based methods without animal or clinical trials.Despite strong in vitro findings,in vivo studies are needed to confirm licorice glycoside E’s therapeutic potential and understand the microRNA-149-5p-MMP9 dynamics in real conditions.

Licorice:comprehensive review of its chemical composition,pharmacodynamics,and medicinal value

Licorice,a perennial herb of Leguminosa,is one of the oldest and most widely used herbal medicines worldwide.Its distinct sweet flavor and rich medicinal value make it an integral component of traditional Chinese medicine(TCM)formulations,which continue to be widely employed.The main chemical constituents of licorice include triterpenoid saponins,flavonoids,and polysaccharides.Experimental and clinical studies have demonstrated that various extracts and pure compounds derived from licorice exhibit a wide range of pharmacological properties including anti-inflammatory,antioxidant,antimicrobial,antiviral,antitumor,immune-regulatory,and neuroprotective activities.The bioactive constituents of licorice offer therapeutic benefits for cardiovascular and cerebrovascular diseases,diabetes mellitus,and liver disorders.This comprehensive review discusses the primary chemical constituents of licorice and their pharmacological activities,describes in vivo and in vitro models employed for studying licorice,and its potential targets and mechanisms of action.Furthermore,we discuss the toxicological profile,side effects,dosage recommendations,and clinical applications of licorice.This review aims to establish a foundation for further research on the safe and effective utilization of licorice while facilitating an in-depth exploration of its properties and fostering the development of novel therapeutic agents.

Studies on a Traditional Chinese Medical Prescription Ma Xing Shi Gan Tang by Bio-coordination Chemistry

本文报道 pH 电位滴定法测出麻黄碱与生石膏中的 Cu^(2+)、Fe^(2+)、Zn^(2+)、Mn^(2+)离子形成的 ML 及 ML_2型络合物的稳定常数。滴定曲线显示甘草酸与上述四种金属离子也能分别形成不同稳定程度的络合物。用麻黄碱加甘草酸和上述四种金属离子配制成的模拟麻杏石甘汤和单纯生石膏煎剂与麻性石甘汤一样均有使家兔退热的效果,与对照组比较均有显著性差异(P<0.05);而用麻黄碱加甘草酸配制成的溶液无退热效果。作者认为应从生物—配位无机化学的多金属—多配体体系的观点去研究某些传统中医药方剂。

MICROWAVE-ASSISTED EXTRACTION OF GLYCYRRHIZIC ACID FROM LICORICE ROOT-EFFECT OF THE PROPERTY OF SOLUTION ON EXTRACTION OF GA

The property of extraction solution is an important factor influencing the extraction efficiency. In the present work, the effect of the property of solution on extraction of GA was studied, which including the concentration of ethanol, ammonia and cation (M+), pH of extraction solution, different kinds of organic solvent etc. The results show that 50%-60%(v/v) ethanol can reach high percentage extraction of GA. If 1% (v/v) ammonia solution was added into 60%(v/v) ethanol, the percentage extraction can be increased from 2.0% to 2.31%. Without ammonia, 50mmol/L [M+] (M+ = K+, NH4+) was added into 60%(v/v) ethanol, percentage extraction of GA can reach about 2.26%. If pH of solution (60% ethanol) was adjust to pH=4.0, it can reach high percentage extraction. If pH of solution (60% ethanol + 50mmol [M+], pH=6.1) was adjust tO PH=4.0, especially M+ is K+ or NH4+, it can reach almost same extraction efficiency as that of 1% ammonia solution + 60% ethanol, and the operation environment can be greatly improved.

Arbuscular mycorrhizal fungal colonization of Glycyrrhiza glabra roots enhances plant biomass,phosphorus uptake and concentration of root secondary metabolites

Arbuscular mycorrhizal （AM） fungi penetrate the cortical cells of the roots of vascular plants, and are widely distributed in soil. The formation of these symbiotic bodies accelerates the absorption and utilization of min- eral elements, enhances plant resistance to stress, boosts the growth of plants, and increases the survival rate of transplanted seedlings. We studied the effects of various arbuscular mycorrhizae fungi on the growth and devel- opment of licorice （Glycyrrhiza glabra）. Several species of AM, such as Glomus mosseae, Glomus intraradices, and a mixture of fungi （G. mosseae, G intraradices, G. cladoideum, G microagregatum, G caledonium and G. etunica- tum） were used in our study. Licorice growth rates were determined by measuring the colonization rate of the plants by the fungi, plant dry biomass, phosphorus concentration and concentration of secondary metabolites. We estab- lished two cloned strains of licorice, clone 3 （C3） and clone 6 （C6） to exclude the effect of genotypic variations. Our results showed that the AM fungi could in fact increase the leaf and root biomass, as well as the phosphorus con- centration in each clone. Furthermore, AM fungi significantly increased the yield of certain secondary metabolites in clone 3. Our study clearly demonstrated that AM fungi play an important role in the enhancement of growth and development of licorice plants. There was also a significant improvement in the secondary metabolite content and yield of medicinal compounds from the roots.

Licorice Extracts Attenuate Nephrotoxicity Induced by Brucine Through Suppression of Mitochondria Apoptotic Pathway and STAT3 Activation

Licorice,one of the most widely used medicinal herbs in East Asia,has effects such as anti-inflammation,antioxidant,and detoxifying.This study aimed to evaluate the protective effect of licorice on brucine-induced nephrotoxicity.Sprague Dawley rats were administered with brucine intraperitoneally for 7 consecutive days with or without treatment with licorice.The content of blood urea nitrogen and creatinine in serum,the activities of superoxide dismutase and content of glutathione,malonaldehyde in kidney tissue were detected.Hematoxylin-eosin staining was employed to observe the histopathological changes of kidney.The expression and phosphorylation levels of protein were evaluated by Western blotting and immunohistochemical analysis.The results illustrated that treatment with licorice extracts(LE)significantly protected against the brucineinduced nephrotoxicity by reducing the content of blood urea nitrogen and serum creatinine,attenuating pathologic damage.The unbalance of oxidative stress was repaired by LE via increasing the level of glutathione,promoting the activities of superoxide dismutase and decreasing the content of malonaldehyde.In addition,LE overturned the influence of brucine on apoptosis-related protein and signal transducer and activator of transcription-3(STAT3)activation.Taken together,these data demonstrate that licorice may attenuate brucine-induced nephrotoxicity via inactivation of oxidative stress and mitochondrial-mediated apoptosis pathway.More importantly,the renoprotective effects may be mediated,at least partly,by preventing the activation of STAT3 protein.

内蒙古鄂尔多斯高原的甘草资源及速生繁殖

鄂尔多斯高原位于内蒙古自治区的西南部,这里地形复杂,境内多沙漠和丘陵山地,主要有库布齐和毛乌素两大沙地,海拔平均在1000—1500m之间。鄂尔多斯高原土地贫瘠,干旱少雨,风大沙多,气候干燥,属于半干旱—干旱典型草原与荒漠草原区。无霜期较短,光照时间长,有效积温高,降雨集中,水热同季。

Development and evaluation of a safe and effective sugar-free herbal lollipop that kills cavity-causing bacteria

Dental caries （tooth decay） is caused by a specific group of cariogenic bacteria, like Streptococcus mutans, which convert dietary sugars into acids that dissolve the mineraI in tooth structure. Killing cariogenic bacteria is an effective way to control or prevent tooth decay. In a previous study, we discovered a novel compound （Glycyrrhizol A）, from the extraction of licorice roots, with strong antimicrobial activity against cariogenic bacteria. In the current study, we developed a method to produce these specific herbal extracts in large quantities, and then used these extracts to develop a sugar-free lollipop that effectively kills cariogenic bacteria like Streptococcus mutans. Further studies showed that these sugar-free lollipops are safe and their antimicrobial activity is stable. Two pilot human studies indicate that a brief application of these lollipops （twice a day for ten days） led to a marked reduction of cariogenic bacteria in oral cavity among most human subjects tested. This herbal lollipop could be a novel tool to promote oral health through functional foods.

Improving the Antibacterial Property of Chitosan Hydrogel Wound Dressing with Licorice Polysaccharide

A series of hydrogels with different ratios of chitosan and licorice polysaccharide(LP)were prepared by crosslinking to different concentrations of genipin(gp).They were characterized by FTIR(Fourier transform infrared spectroscopy),SEM(Scanning electron microscope),swelling ratio,rheological measurements,degradation with time,cytotoxicity,and antibacterial efficacy.Results show that the hydrogels have porous structures.With an increase in LP content,the swelling rate grows in the early stage of immersion in buffer and drops later.The swelling ratio ranged from 986%to 1677%,and stiffness varied from 777 Pa to 1792 Pa.The addition of LP reduced the mechanical strength and delayed gelation and degradation of the hydrogels.However,the most important discovery was that gp increases the viability of NIH 3T3 cells from 94%to 137%,and LP raises the bacteriostatic efficacy from 51%to 78%.Hydrogels synthesized from 1%genipin,3%chitosan,and 4%licorice polysaccharide showed the best antibacterial and fibroblast proliferation promoting activities.They exhibited moderate swelling and degradation rates over time,while being more suitable to affect healing of chronic wound infections.These results provide a new strategy to improve the antibacterial effectiveness and cyto-compatibility of chitosan hydrogels with water soluble active LPs from Glycyrrhiza that derive from traditional Chinese medicine.

Elucidating the interaction of kansui and licorice by comparative plasma/tissue metabolomics and a heatmap with relative fold change

Although compatibility is highly advocated in traditional Chinese medicine (TCM), inappropriate combination of some herbs may reduce the therapeutic action and even produce toxic effects. Kansui and licorice, one of TCM “Eighteen Incompatible Medicaments”, are the most representative cases of improper herbal combination, which may still be applied simultaneously under given conditions. However, the potential mechanism of their compatibility and incompatibility is unclear. In the present study, two different ratios of kansui and licorice, representing their compatibility and incompatibility respectively, were designed to elucidate their interaction by comparative plasma/tissue metabolomics and a heatmap with relative fold change. As a result, glycocholic acid, prostaglandin F2a, dihydroceramide and sphinganine were screened out as the principal alternative biomarkers of compatibility group;sphinganine, dihydroceramide, arachidonic acid, leukotriene B4, acetoacetic acid and linoleic acid were those of incompatibility group. Based on the values of biomarkers in each tissue, the liver was identified as the compatible target organ, while the heart, liver, and kidney were the incompatible target organs. Furthermore, important pathways for compatibility and incompatibility were also constructed. These results help us to better understand and utilize the two herbs, and the study was the first to reveal some innate characters of herbs related to TCM “Eighteen Incompatible Medicaments”.

Performance, Immune, and Carcass Characteristics of Broiler Chickens as Affected by Thyme and Licorice or Enzyme Supplemented Diets

Numbers of 300 day-old broiler chickens through a CRD design with 5 treatments, 3 replicates and 20 chicks in each pen were used to evaluate the effect of thyme (T), licorice (L), thyme + licorice (TL), and enzyme supplemented (E) diets on performance, immune and carcass characteristics. According to the results, performance traits, immune indices, and carcass traits in herbal medicine and enzyme supplemented diets were improved significantly than control diet (P < 0.05). Weight gain and FCR in T and E groups were significantly higher and lower than other groups respectively (P < 0.05). Internal organs such as abdominal fat and liver weight as indicators of lipogenesis rate were decreased in T, L, and TL diets than control or E diet significantly (P < 0.05). Immune organs such as burse and spleen weight as indicators of immune situation were increased in TL diet than other treatments significantly (P < 0.05). These findings indicated that thyme and licorice singly or in combination as organic herbal medicine can affect performance, carcass and immune characteristics. Also an improved immune organ such as burse or spleen in this study indicates that this herbal medicine can promote the immune situation and efficacy of health and livability.

Molecular Cloning and Expression Analysis of an Actin Gene from Chinese Licorice(Glycyrrhiza uralensis Fisch)

A PCR-based homologous cloning strategy was used to identify an actin gene from the roots of Chinese licorice（Glycyrrhiza uralensis Fisch）. Results of sequence analysis indicate that a 1137 bp cDNA with an open reading frame encoding 377 amino acids, actin ortholog, GuActin, was successfully cloned and characterized（GenBank accession No. EU190972）. Thus far, GuActin is the first actin of Chinese licorice that has been identified at a molecular level. Analysis by Northern blot shows that GuActin was expressed strongly in the roots, particularly in radicles than in stems and leaves. These results suggest that GuActin may be a member of the vegetative subfamily of the actin family.

Novel Inhibitory Effects of Glycyrrhizic Acid on the Accumulation of Advanced Glycation End Product and Its Receptor Expression

Beneficial effects of glycyrrhizic acid(GA),a bioactive extract of licorice root,in the prevention of metabolic syndrome have been consistently reported while advanced glycation end products(AGE)and receptor for advanced glycation end product(RAGE)are the leading factors in the development of diabetes mellitus.The aim of this study was to investigate the effects of GA on the AGE-RAGE axis using high-fat/high-sucrose(HF/HS)diet-induced metabolic syndrome rat models.Twenty four male Sprague–Dawley rats were randomly assigned into three groups for 4 weeks:(1)Group A,normal diet with standard rat chow;(2)Group B,HF/HS diet;(3)Group C,HF/HS diet and oral administration of 100 mg/kg GA per day.The results showed that HF/HS diet elevated the fasting blood glucose level and insulin resistance index which was prevented by GA supplementation.GA treatment significantly lowered the circulating AGE independent of its glucose-lowering effect.HF/HS diet also triggered RAGE upregulation in the abdominal muscles while GA administration downregulated RAGE expression in the abdominal muscles,aorta and subcutaneous adipose tissues.In conclusion,HF/HS diet could cause glucose intolerance,insulin resistance and upregulation of RAGE expression while GA ameliorated the metabolic dysregulation besides exhibiting inhibitory effects on the AGE-RAGE axis.

Licorice and its main components on the treatment of diabetes and its complications:A review of in vivo and in vitro studies

Licorice is one of the oldest herbal medicines for its various ethno pharmacological uses.In both Asian and European countries,it has been recorded for treatment of inflammatory diseases.A large number of ingredients have been isolated from licorice,including triterpene saponins and flavonoids,which are normally being considered to be the main biologically active components.In the last decade,licorice has been proved exert anti-diabetic effect in various in vivo and in vitro models of diabetes mellitus.Furthermore,licorice can also antagonize all sorts of diabetes complications,including diabetic nephropathy,atherosclerosis,diabetic retinopathy and neuropathy.Except anti-inflammation,licorice and its active components show anti-diabetic effects by improving insulin resistance and increasing insulin secretion,regulating lipid metabolism,and anti-oxidation.The useful effects of licorice and its active components are due to regulating different pathways and proteins,including NF-κB,AMPK,insulin signaling pathway,MAPK,etc.In this review,we provide an overview of the beneficial effects and related molecular mechanism of licorice and its effective components on improving diabetes and its complications.

Alginate encapsulation in Glycyrrhiza glabra L.with phyto-chemical profiling of root extracts of in vitro converted plants using GC-MS analysis

Objective: To investigate the conversion potential of alginate encapsulated nodes of Glycyrrhiza glabra with phyto-chemical evaluation of root extract of field transferred plants.Methods: The excised axenic nodal segments were encapsulated in alginate matrix planted on Murashige and Skoog(1962) medium with different supplementation and formulations of PGRs. The two year old field transferred plants were evaluated for phytocompounds analysis using GC-MS technique.Results: Varied responses were observed during the study, maximum conversion95.83% ± 2.40% was obtained in these encapsulates when planted on MS medium containing 2.5 m M Kinetin and 0.5 m M a-Naphthalene acetic acid, which eventually developed into complete plantlets in a single step. Further, GC-MS analysis showed the presence of different phyto-compounds in the methanolic root extracts of in vitro converted plants. The results obtained revealed the presence of about 47 phyto-compounds along with various potential bioactive compounds useful for industrial and pharmaceutical purposes.Conclusions: This study investigates high frequency regeneration and conversion of Glycyrrhiza glabra in a single step in short time. Also, the in vitro raised plants are analysed for bioactive compounds after field transfer, which shows the presence of numerous compounds useful for commercial and pharmacological purposes.

Black licorice ingestion:Yet another confounding agent in patients with melena

We describe an 80-year-old woman with atrial fibrillation,anti-coagulated with warfarin,who on two separate occasions developed black tarry stools and an elevated international normalized ratio(INR) after eating a pound of Black Licorice.During her most recent episode,her hematocrit was 14(baseline 34) and her INR was 5.5(baseline 2.1).She was advised to restrict licorice consumption,and a follow-up INR two weeks later was 1.2.Black Licorice is derived from the root of the plant,Glycyrrhiza glabra.The components of its extract inhibit the P450 system enzymes that metabolize Warfarin,inhibit thrombin,and prolong fibrinogen clotting times.Hence,the anti-thrombotic activity and inhibition of warfarin metabolism might synergistically amplify anticoagulation.The presence of Black Licorice in the stool can also mimic melena and confound its clinical presentation.Health care providers should caution patients who are at risk for bleeding or on warfarin to avoid black licorice due to an elevated risk of gastroint estinal bleeding.

THE EFFECT OF VIOLENT DIGGING OF LICORICE RESOURCES ON THE ENVIRONMENT OF ARID AREA

Licorice (Glycyrrhira) is one of the important medical resources in arid area. It is dis-tributed between 30 and 55 degrees north latitude, especially in arid area 40 degrees northlatitude. There are about nine or ten species of licorice in our country. Not only it has ahigh economical value, but also it is an important sand-control plant for

Pressured Microwave-assisted Hydrolysis of Crude Glycyrrhizic Acid for Preparation of Glycyrrhetinic Acid

A pressured microwave-assisted hydrolysis （PMAH） technique has been developed for hydrolyzing the crude glycyrrhizic acid （GA） extracted from licorice root to prepare glycyrrhetinic acid （GRA）. In order to optimize the efficiency of PMAH, several experimental parameters were investigated, including liquid-solid ratio, hydrolysis time, sulfuric acid concentration and hydrolysis temperature. The optimized hydrolysis conditions were as follows：pressured microwave-assisted hydrolysis of crude GA for 21 min （taking 15 min to reach 150 ℃, and holding it for 6 rain） at 150 ℃ （at a radiation power of 450 W） in 3%-5% sulfuric acid solution with the liquid-solid （ml.g-1 crude GA） ratio of 25 ： 1. As a result of the considerable saving in time and higher product yields （up to 90%）, PMAH was proved more effective than conventional methods.

Promoting effect of licorice extract on induction of β-glucuronidase in Penicillium purpurogenum Li-3

PeniciUium purpurogenum Li-3, a fungus producing β-glucuronidase （PGUS）, can con- vert glycyrrhizin （GL） to glycyrrhetinic acid monoglucuronide （GAMG） when grown in medium with GL as the sole carbon source. In order to improve the conversion rate of GL and the yield of GAMG, licorice extract （LE） was added as an inducer to enhance the production of GAMG by the PGUS. In this work, the influence of LE on the conversion rate of GL to GAMG was studied. When the Penicil- lium purpurogenum Li-3 was grown in the medium containing LE and GL （ concentration ratio of LE to GL was 2： 3）, the conversion rate of GL was 84. 12% with 38. 18% increase and the yield of GAMG was 80. 47% with 37. 18% increase, comparing with to the medium only containing GL at 48 h. The enzyme activity of ^-glucuronidase was also enhanced from 22. 4 U/mL to 82.3 U/mL, which in- creased up to about 3. 67 fold. The results showed that LE could significantly improve the induced expression level of PGUS.

Estimation of multivariate detection limits of four quality parameters in licorice using MEMS–NIR spectrometry coupled with two sampling accessories

In this work,multivariate detection limits(MDL)estimator was obtained based on the microelectro-mechanical systems–near infrared(MEMS–NIR)technology coupled with two sampling accessories to assess the detection capability of four quality parameters(glycyrrhizic acid,liquiritin,liquiritigenin and isoliquiritin)in licorice from di®erent geographical regions.112 licorice samples were divided into two parts(calibration set and prediction set)using Kennard–Stone(KS)method.Four quality parameters were measured using high-performance liquid chromatography(HPLC)method according to Chinese pharmacopoeia and previous studies.The MEMS–NIR spectra were acquired from¯ber optic probe(FOP)and integrating sphere,then the partial least squares(PLS)model was obtained using the optimum processing method.Chemometrics indicators have been utilized to assess the PLS model performance.Model assessment using chemometrics indicators is based on relative mean prediction error of all concentration levels,which indicated relatively low sensitivity for low-content analytes(below 1000 parts per million(ppm)).Therefore,MDL estimator was introduced with alpha error and beta error based on good prediction characteristic of low concentration levels.The result suggested that MEMS–NIR technology coupled with fiber optic probe(FOP)and integrating sphere was able to detect minor analytes.The result further demonstrated that integrating sphere mode(i.e.,MDL0:05;0:05,0.22%)was more robust than FOP mode(i.e.,MDL0:05;0:05,0.48%).In conclusion,this research proposed that MDL method was helpful to determine the detection capabilities of low-content analytes using MEMS–NIR technology and successful to compare two sampling accessories.