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.

Inhibitory Effect of Flavonoid Glycosides from Chlorophytum comosum on Nasopharyngeal Carcinoma 5-8F Cells and Its Mechanism

[Objectives]To study the inhibitory activity of two flavonoid glycosides isolated from Chlorophytum comosum Laxum R.Br on human nasopharyngeal carcinoma(NPC)cell line 5-8F in vitro and its mechanism.[Methods]The flavonoid glycosides were isolated and purified from the ethanol alcoholic extract of the roots of Liliaceae plant Chlorophytum comosum by silica gel column chromatography,macroporous resin column chromatography,Sephadex LH-20,and reverse column chromatography(ODS).The inhibitory activity of flavonoid glycosides on human nasopharyngeal carcinoma cells was analyzed by CCK-8 method,and the potential mechanism was preliminarily analyzed by molecular docking.[Results]Two flavonoid glycosides were identified as isovitexin 2″-0-rhamnoside and 7-2″-di-O-β-glucopyranosylisovitexin.Two flavonoid glycosides showed promising inhibitory effect on human nasopharyngeal carcinoma cell line 5-8F,with IC_(50) values of 24.8 and 27.5μmol/L,respectively.Molecular docking results showed that the potential targets of two flavonoid glycosides include CyclinD1,Bcl-2β-Catenin,ILK,TGF-β,in addition,two glycosides showed higher predicted binding affinity towards CyclinD1,which verifies the cytotoxicity of the two compounds on human nasopharyngeal carcinoma cell line 5-8F in vitro.[Conclusions]Two flavonoid glycosides are the active molecules in Chlorophytum comosum that can inhibit the proliferation of human nasopharyngeal carcinoma cells,and have the potential to be used in the research and development of anti nasopharyngeal carcinoma drugs.

Single-cell transcriptome analysis uncovers underlying mechanisms of acute liver injury induced by tripterygium glycosides tablet in mice

Tripterygium glycosides tablet(TGT),the classical commercial drug of Tripterygium wilfordii Hook.F.has been effectively used in the treatment of rheumatoid arthritis,nephrotic syndrome,leprosy,Behcet's syndrome,leprosy reaction and autoimmune hepatitis.However,due to its narrow and limited treatment window,TGT-induced organ toxicity(among which liver injury accounts for about 40%of clinical reports)has gained increasing attention.The present study aimed to clarify the cellular and molecular events underlying TGT-induced acute liver injury using single-cell RNA sequencing(scRNA-seq)technology.The TGT-induced acute liver injury mouse model was constructed through short-term TGT exposure and further verified by hematoxylin-eosin staining and liver function-related serum indicators,including alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase and total bilirubin.Using the mouse model,we identified 15 specific subtypes of cells in the liver tissue,including endothelial cells,hepatocytes,cholangiocytes,and hepatic stellate cells.Further analysis indicated that TGT caused a significant inflammatory response in liver endothelial cells at different spatial locations;led to marked inflammatory response,apoptosis and fatty acid metabolism dysfunction in hepatocytes;activated hepatic stellate cells;brought about the activation,inflammation,and phagocytosis of liver capsular macrophages cells;resulted in immune dysfunction of liver lymphocytes;disturbed the intercellular crosstalk in liver microenvironment by regulating various signaling pathways.Thus,these findings elaborate the mechanism underlying TGT-induced acute liver injury,provide new insights into the safe and rational applications in the clinic,and complement the identification of new biomarkers and therapeutic targets for liver protection.

Phenylethanoid glycosides from traditional Mongolian medicine Cymbaria daurica alleviate alloxan-induced INS-1 cells oxidative stress and apoptosis

Cymbaria daurica L. is a well-known traditional Mongolian medicine, which has been used to treat diabetesrelated conditions characterized by persistent thirst and hunger, copious urination, and weight loss. We aimed to investigate the protective effects of C. daurica extracts and phenylethanoid glycosides including verbascoside and isoacteoside on INS-1 cells. We discovered phenylethanoid glycosides from n-butanol extract with large content through extraction and separation. We continue to study the protective effects of phenylethanoid glycosides including verbascoside and isoacteoside on INS-1 cells. INS-1 cells were treated with C. daurica, cell viability assay, RNA-seq technology, superoxide dismutase activity and malonaldehyde content, quantitative real time-PCR and Western blot analysis were used to study the protective effects of C. daurica. Cell viability assay resulted that n-butanol extract and verbascoside, isoacteoside showed protective effects of C. daurica. According to the RNA-seq technology to identify the differentially expressed genes in INS-1 cells, the pathway of gene enrich the protective effect of C. daurica on oxidative stress. SOD activity and the content of MDA indicated that C. daurica could enhance the antioxidant capacity of INS-1 cells. Further investigation indicated C. daurica alleviate oxidative stress by inhibiting INS-1 cell apoptosis. C. daurica may play an anti-diabetic role by inhibiting islet cell apoptosis.

The effect and mechanism of stilbene glycosides on improving neuronal injury in Alzheimer's disease rats by regulating ASK/MKK7/JNK pathway

Objective:To investigate the mechanism of action of tetrahydroxy stilbene glycosides(TSG)in ameliorating neuronal damage in Alzheimer's disease rats by regulating MKK7 and JNK kinases.Methods:A total of 24-month-old 42 SD rats were randomly selected for the experiment in 7 groups:normal group,sham-operated group,model group,positive drug group,low,medium and high dose TSG group at 0.033 g/kg,0.1 g/kg,0.3 g/kg.The Model Group and the TSG groups were established by stereotaxic Aβ25-35 solution.After 28 days,the model rats were selected by passive avoidance test.After screening,each dosage group of TSG and positive drug group was given intragastrically according to the corresponding dosage,and the experiment was carried out after 28 days.The pathological changes of hippocampal CA1 region were observed by tissue staining,and the amount of MKK7 and JNK proteins and the expression content of MKK7 and JNK mRNA by histochemical method of protein,and qRTPCR assay.Results:(1)He staining observation:Compared with the normal group and the sham-operated group,the number of nerve cells in the model group decreased and arranged irregularly,the cell membrane shrank,and the nucleus deformed and dissolved.The number of neurons in the positive drug group and TSG Group also increased significantly,the order is also relatively well.(2)From the results of the Tunel staining experiments:the positive apoptotic cells in the model group were higher than control group and sham-operated group,positive drug group and TSG drugs group was significantly smaller than that in the model group(P<0.05).(3)Compared with the control group and the Virtual Operation Group,the MKK7 and JNK protein concentrations in the brain of the model group were increased(P<0.05)by data analysis of immunohistochemistry:Compared with the model group,the protein expression of positive drug and TSG each dose group were reduced(P<0.05).(4)The results of QRTPCR data showed that the levels of MKK7 and JNK mRNA in the brain tissue of the model group were increased compared with the normal group and sham-operated groups(P<0.05).Conclusion:Stilbene glycoside has a certain effect on neuronal injury and repair which may be related to the changes of mRNA transcription and protein expression of MKK7 and JNK kinases.

川赤芍总苷提取工艺优化、组成和抗氧化活性分析

探究超声波辅助乙醇提取川赤芍总苷,通过单因素和响应面优化提取工艺,采用高效液相色谱(HPLC)法检测总苷组成,并通过体外抗氧化试验评价总苷的抗氧化活性。结果表明:最佳提取工艺为超声功率175 W、超声时间42 min、乙醇浓度70%、液料倍数30(mL/g),此工艺条件下提取的川赤芍总苷得率为(5.86±0.32)%。HPLC分析总苷共检测到21个特征峰,其中没食子酸0.63 mg/g、氧化芍药苷11.28 mg/g、儿茶素18.64 mg/g、芍药内酯苷5.82 mg/g、芍药苷54.82 mg/g、苯甲酰芍药苷15.74 mg/g、丹皮酚原苷0.98 mg/g,共占总质量的95.35%。总苷清除DPPH·、ABTS+·和·OH-的IC50值分别为0.78、1.13、0.98 mg/mL,总苷的抗氧化活性强弱与浓度呈现较好的正相关。

山银花茎叶化学成分及其抗炎活性研究

目的对山银花茎叶化学成分进行分离和结构鉴定,并进行体外抗炎活性评价。方法山银花茎叶80%甲醇提取物采用Diaion HP20SS、Sephadex LH-20、高速逆流色谱、反相半制备液相色谱等进行分离纯化,根据理化性质及波谱数据鉴定所得化合物的结构。通过测定单体化合物抑制LPS诱导小鼠巨噬细胞RAW264.7释放NO水平评价其抗炎活性。结果从山银花茎叶中分离得到13个化合物,分别鉴定为苄醇-O-β-D-吡喃葡萄糖-(1→6)-β-D-吡喃葡萄糖苷(1)、獐牙菜苷(2)、表断马钱子苷半缩醛内酯(3)、马钱子苷半缩醛内酯(4)、裂环马钱苷(5)、断氧化马钱苷(6)、裂马钱素二甲基乙缩醛(7)、绿原酸甲酯(8)、芹菜素-7-O-β-D-葡萄糖苷(9)、木犀草素-7-O-β-D-葡萄糖苷(10)、野漆树苷(11)、木犀草素-7-O-α-L-吡喃阿拉伯糖(1→6)-β-D-吡喃葡萄糖苷(12)、忍冬苷(13);化合物2~8、11~13可抑制LPS诱导RAW264.7细胞释放NO。结论化合物1首次从忍冬属植物中分离得到,化合物3、5、7、9、11~12首次从山银花茎叶中发现。化合物2~8、11~13具有一定的抗炎活性。

何首乌叶不同萃取物的抗炎活性及其化学成分鉴定

目的:研究何首乌叶的成分,为其功能食品的研发提供基础数据。方法:对何首乌叶醇提取物进行有机溶剂萃取,分别得到何首乌叶石油醚、二氯甲烷、乙酸乙酯、正丁醇、水等不同萃取物,并对不同萃取物进行抗炎活性检测,后续通过硅胶柱色谱、C18柱色谱结合半制备液相等方法对抗炎活性较好的成分进行分离、纯化以及结构鉴定,结果:在质量浓度0~100μg/mL范围内,乙酸乙酯萃取物对RAW264.7细胞存活率无明显的抑制作用,且剂量依赖地抑制NO的生成。石油醚和二氯甲烷萃取物在质量浓度0~25μg/mL时,均无明显细胞毒性,并在25μg/mL时,二氯甲烷萃取物对NO的抑制率(51.47%)高于石油醚萃取物的抑制率(43.91%)。从二氯甲烷和乙酸乙酯萃取物中分离、鉴定出11个化合物,分别为2,3,4,6-三羟基苯乙酮-3-O-β-D-glucoside(1)、杨梅苷(2)、槲皮苷(3)、阿福豆苷(4)、β-谷甾醇(5)、正十六烷-5,8,11三烯酸(6)、二十六烷(7)、α-亚麻酸(8)、山柰酚-3-O-芸香糖苷(9)、肉豆蔻酸(10)和槲皮素(11)。结论:何首乌叶具有很好的抗炎活性,化合物2,4,6,7,8,9,10,11均为首次从何首乌中分离得到。

Caffeoyl Glycoside对小鼠免疫细胞功能的影响

本实验室从胡黄连根茎分离到1,6-di-O-caffeoyl-β-D-glucopyranoside单体Caffeoyl Glycoside(CG),体外试验测定其对小鼠免疫细胞功能的影响。通过MTT法检测CG对Balb/c小鼠脾淋巴细胞增殖,NK细胞对K562细胞杀伤活性的影响,以及小鼠腹腔巨噬细胞(PMΦ)能量代谢水平;CG对PMΦ吞噬中性红能力的影响。CG在一定剂量范围内单独或者协同非特异性丝裂原(ConA或LPS)作用均能够显著增强小鼠脾淋巴细胞、PMΦ的增殖能力(P<0.05或P<0.01),CG单独作用就能显著提高NK对K562细胞的杀伤活性(P<0.05或P<0.01)。CG在体外对小鼠主要免疫细胞的增值和功能方面具有显著的促进作用(P<0.05或P<0.01)。研究结果显示CG具有特异性与非特异性的免疫增强作用。

Caffeoyl Glycoside对脾淋巴细胞生长周期及膜表面标志的影响

用流式细胞术测定了从西藏胡黄连分离的Caffeoyl Glycoside(CG)对小鼠脾淋巴细胞生长周期及其膜表面标志CD4+、CD8+的影响.结果表明:CG通过促进脾淋巴细胞G0/G1期向DNA合成期(S期)转化,从而促进脾淋巴细胞增殖,对脾淋巴细胞膜表面标志的影响,是通过CG上调CD4+、CD8+和CD4+CD8+双阳性细胞亚群,即主要是通过提高Th细胞的数量发挥免疫增强作用.

Caffeoyl Glycoside对小鼠免疫细胞功能的影响

本实验室从胡黄连根茎分离到1,6-di-O-caffeoyl-β-D-glucopyranoside单体Caffeoyl Glycoside(CG),体外试验测定其对小鼠免疫细胞功能的影响。通过MTT法检测CG对Balb/c小鼠脾淋巴细胞增殖,NK细胞对K562细胞杀伤活性的影响,以及小鼠腹腔巨噬细胞(PMΦ)能量代谢水平;CG对PMΦ吞噬中性红能力的影响。CG在一定剂量范围内单独或者协同非特异性丝裂原(Con A或LPS)作用均能够显著增强小鼠脾淋巴细胞、PMΦ的增殖能力(P<0.05或P<0.01),CG单独作用就能显著提高NK对K562细胞的杀伤活性(P<0.05或P<0.01)。CG在体外对小鼠主要免疫细胞的增值和功能方面具有显著的促进作用(P<0.05或P<0.01)。研究结果显示CG具有特异性与非特异性的免疫增强作用。

A New Flavanone and a New Flavanone Glycoside from Scutellaria baicalensis

A new flavanone （2S）-5, 7, 2＇, 5＇-tetrahydroxy-flavanone 1 together with a new flavanone glycoside （2S）-5, 7, 2＇, 5＇-tetrahydroxy-flavanone 7-O-13-D-glucopyranoside 2, were isolated from the dry roots of Scutellaria baicalensis. Their structures were elucidated on the basis of spectroscopic data.

红花钓钟柳中新型糖基转移酶的筛选与鉴定

苯乙醇苷类化合物因具有抗氧化、抗肿瘤等生理活性在食品、医药等领域备受关注。糖基转移酶作为生物催化剂可用于各种糖苷产物的制备。为丰富苯乙醇苷类化合物的糖苷多样性,在红花钓钟柳转录组中筛选糖基转移酶候选基因并进行功能鉴定。该研究基于不同植物组织中基因差异表达分析筛选到7条候选基因,将其克隆到pET-MBP表达载体上,并导入大肠杆菌Escherichia.coli Transetta(DE3)异源表达后利用体外酶促反应进行其生物催化活性研究。获得一个新型糖基转移酶UGT712A2,可以催化桂叶苷B C4-OH的葡萄糖基化反应生成新的苯乙醇苷类化合物桂叶苷B-4-葡萄糖苷。同时,UGT712A2也可以催化毛蕊花糖苷、芦丁、胡黄连苷Ⅱ、7-去甲基软木花椒素等化合物的糖基化,具有一定的底物宽泛性。因此,UGT712A2可能是一种潜在的酶工具,用于苯乙醇苷类化合物和其他多种新型糖苷产物的制备,从而发现新的糖苷化合物应用于食品、医药领域。

17-Dehydroxyholothurin A, a New Sulfated Triterpene Glycoside from Sea Cucumber Holothuna impatiens

Abstract： A new sulfated triterpene glycoside named as 17-dehydroxyholothurin A 1 was isolated from the sea cucumber Holothuria impatiens. Its structure was elucidated on the basis of spectroscopic data （2D NMR and MS） and chemical evidence. Compound 1 showed cytotoxic activity against cancer cell lines MKN-28, MFC-7, KB, HL-60 and Hep G2 with IC50 values of 1.98, 4.53, 2.01, 4.69 and 2.80 ×10^-6 tool/L, respectively.

马鞭草苯乙醇苷类成分研究

目的研究马鞭草Verbenae Herba的苯乙醇苷类成分。方法马鞭草80%乙醇提取物采用硅胶、Sephadex LH-20、TLC、半制备HPLC进行分离纯化,根据理化性质及波谱数据鉴定所得化合物的结构。结果从中分离得到9个化合物,分别鉴定为verbofficoside A(1)、肉苁蓉苷D(2)、广防风苷A(3)、毛蕊花糖苷(4)、异毛蕊花糖苷(5)、肉苁蓉苷C(6)、肉苁蓉苷F(7)、去咖啡酰基毛蕊花苷(8)、jionoside C(9)。结论化合物1为新化合物,化合物3、6~9均首次从该植物中分离得到。

A New Disulfated Triterpene Glycoside from the Sea Cucumber Mensamaria intercedens Lampert

A new disulfated triterpene glycoside named intercedenside B was isolated from the sea cucumber Mensamaria intercedens Lampert. The structure was deduced from the spectral data.

A New Pregnane Glycoside from Fermented Leaves of Agave americana

A new minor pregnane glycoside was isolated from the fermented leaves of Agave americana. Its structure was elucidated as (20S)-5a-pregnane-3? 20-diol 20-O--D-glucopyrano- side (1) by spectral methods.

A New Dihydroflavone Glycoside from Glycyrrhiza uralensis

A new dihydroflavone glycoside was isolated from the underground parts of Glycyrrhiza uralensis. Its structure was elucidated as 7-hydroxyl-4'-O-β-D-(6'-O-α-hydroxylpropionyl) glucopyranosyl dihydroflavone by spectral methods.

Protective mechanism of Paeoniae Radix Alba against chemical liver injury based on network pharmacology,molecular docking,and in vitro experiments

Objective:To explore and validate the potential targets of Paeoniae Radix Alba(P.Radix,Bai Shao)in protecting against chemical liver injury through network pharmacology,molecular docking technology,and in vitro cell experiments.Methods:Network pharmacology was used to identify the common potential targets of P.Radix and chemical liver injury.Molecular docking was used to fit the components,which were subsequently verified in vitro.A cell model of hepatic fibrosis was established by activating hepatic stellate cell(HSC)-LX2 cells with 10 ng/mL transforming growth factor-β1.The cells were exposed to different concentrations of total glucosides of paeony(TGP),the active substance of P.Radix,and then evaluated using the cell counting kit-8 assay,enzyme-linked immunosorbent assay,and western blot.Results:Analysis through network pharmacology revealed 13 key compounds of P.Radix,and the potential targets for preventing chemical liver injury were IL-6,AKT serine/threonine kinase 1,jun protooncogene,heat shock protein 90 alpha family class A member 1(HSP90AA1),peroxisome proliferator activated receptor gamma(PPARG),PTGS2,and CASP3.Gene Ontology(GO)enrichment analysis indicated the involvement of response to drugs,membrane rafts,and peptide binding.Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis revealed that the main pathways involved lipid and atherosclerosis and chemical carcinogenesis-receptor activation.Paeoniflorin and albiflorin exhibited strong affinity for HSP90AA1,PTGS2,PPARG,and CASP3.Different concentrations of TGP can inhibit the expression of COL-I,COL-III,IL-6,TNF-a,IL-1β,HSP-90a,and PTGS2 while increasing the expression of PPAR-γand CASP3 in activated HSC-LX2 cells.Conclusion:P.Radix primarily can regulate targets such as HSP90AA1,PTGS2,PPARG,CASP3.TGP,the main active compound of P.Radix,protects against chemical liver injury by reducing the inflammatory response,activating apoptotic proteins,and promoting the apoptosis of activated HSCs.