Zuogui Jiangtang Jieyu Formula ameliorating hippocampal neuronal apoptosis in diabetic rats with depression by inhibiting JNK signaling pathway

Objective To investigate the effect of Zuogui Jiangtang Jieyu Formula(左归降糖解郁方,ZJJF)on hippocampal neuron apoptosis in diabetic rats with depression and to ascertain whether its mechanism involves the regulation of JNK signaling pathway.Methods(i)A total of 72 specific pathogen-free(SPF)grade male Sprague Dawley(SD)rats were randomly divided into six groups,with 12 rats in each group:control,model,metformin(Met,0.18 g/kg)+fluoxetine(Flu,1.8 mg/kg),and the high-,medium-,and low-ZJJF dosages(ZJJF-H,20.52 g/kg;ZJJF-M,10.26 g/kg;ZJJF-L,5.13 g/kg)groups.All groups except control group were injected once via the tail vein with streptozotocin(STZ,38 mg/kg)combined with 28 d of chronic unpredictable mild stress(CUMS)to establish diabetic rat models with depression.During the CUMS modeling period,treatments were administered via gavage,with control and model groups receiving an equivalent volume of distilled water for 28 d.The efficacy of ZJJF in reducing blood sugar and alleviating depression was evaluated by measuring fasting blood glucose,insulin,and glycated hemoglobin levels,along with behavioral assessments,including the open field test(OFT),forced swim test(FST),and sucrose preference test(SPT).Hippocampal tissue damage and neuronal apoptosis were evaluated using hematoxylin-eosin(HE)staining and terminal deoxynucleotidyl transferase-mediated dUTP nickend labeling(TUNEL)staining.Apoptosis-related proteins Bax,Bcl-2,caspase-3,and the expression levels of JNK/Elk-1/c-fos signaling pathway were detected using Western blot and real-time quantitative polymerase chain reaction(RT-qPCR).(ii)To further elucidate the role of JNK signaling pathway in hippocampal neuronal apoptosis and the pharmacological effects of ZJJF,an additional 50 SPF grade male SD rats were randomly divided into five groups,with 10 rats in each group:control,model,SP600125(SP6,a JNK antagonist,10 mg/kg),ZJJF(20.52 g/kg),and ZJJF(20.52 g/kg)+Anisomycin(Aniso,a JNK agonist,15 mg/kg)groups.Except for control group,all groups were established as diabetic rat models with depression,and treatments were administered via gavage for ZJJF and intraperitoneal injection for SP6 and Aniso for 28 d during the CUMS modeling period.Behavioral changes in rats were evaluated through the OFT,FST,and SPT,and hippocampal neuron damage and apoptosis were observed using HE staining,Nissl staining,TUNEL staining,and transmission electron microscopy(TEM).Changes in apoptosis-related proteins and JNK signaling pathway in the hippocampal tissues of rats were also analyzed.

Therapeutic Effect of Intragastric Administration of Jiangtang Shuxin Recipe Suspension on Diabetic Heart Failure in Rats and Its Mechanism

[Objectives]To observe the therapeutic effect of intragastric administration of Jiangtang Shuxin recipe on diabetic heart failure(DHF)in rats and to explore its mechanism.[Methods]Fifty SD rats were randomly divided into five groups,with 10 rats in each group.DHF models were prepared in the low-dose group,high-dose group,Western medicine group,and model group except the control group.Rats in the low-dose and high-dose groups were given 1.0 and 1.5 g/(kg·d)Jiangtang Shuxin recipe suspension by gavage,respectively.Rats in the Western medicine group were given gliquidone and benazepril by gavage for 2 months,and were fed with high-fat diet.Rats in the control group were fed with ordinary diet.Fasting blood glucose(FBG),serum triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),C-reactive protein(CRP),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),creatine kinase isoenzyme(CK-MB),lactate dehydrogenase(LDH),pathological morphology of myocardial tissue,NF-κB p65 protein and IκBαprotein were compared among groups.[Results]Compared with the control group,the levels of FBG,serum TG,LDL-C,CRP,IL-6,TNF-α,CK-MB and LDH increased,while the level of serum HDL-C decreased.The myocardial tissue was seriously damaged,and the expression of NF-κB p65 protein increased,while the expression of IκBαprotein decreased in the other four gruops(all P<0.05).Compared with the model group,the levels of FBG,serum TG,LDL-C,CRP,IL-6,TNF-α,CK-MB and LDH decreased,while the serum HDL-C level increased.The myocardial tissue damage was alleviated,and the expression of NF-κB p65 protein decreased,while the expression of IκBαprotein increased in the low-dose group,high-dose group and Western medicine group(all P<0.05).Compared with the Western medicine group,the levels of FBG,serum TG,LDL-C,CRP,IL-6,TNF-α,CK-MB and LDH decreased,and the level of serum HDL-C increased in the high-dose group(all P<0.05).[Conclusions]Jiangtang Shuxin recipe has a therapeutic effect on DHF in rats,with the best effect in the high-dose group.It can not only alleviate high glucose and high fat state,but also reduce myocardial injury and inflammation,and improve the pathological morphology of myocardial cells.The mechanism may be related to its inhibition of NF-кB signaling pathway.

Protective effects of Zuogui Jiangtang Jieyu Formula on hippocampal neurons in rats of diabetes complicated with depression via the TRP/KYN metabolic pathway

Objective To explore the protective effects and mechanism of Zuogui Jiangtang Jieyu Formula(左归降糖解郁方,ZGJTJYF)on hippocampal neurons in rats of diabetes complicated with depression(DD)via the TRP/KYN metabolic pathway.Methods(i)In vivo experiments:60 specified pathogen free(SPF)grade male Sprague-Dawley(SD)rats were randomly divided into six groups with 10 rats in each groups:control,DD model,positive(1.8 mg/kg fluoxetine+0.18 g/kg metformin),high-dose ZGJTJYF(ZGJTJYFH,40.500 g/kg ZGJTJYF),middle-dose ZGJTJYF(ZGJTJYF-M,20.250 g/kg ZGJTJYF),and lowdose ZGJTJYF(ZGJTJYF-L,10.125 g/kg ZGJTJYF)groups.Except for the control group,other groups were established DD model by high-fat emulsion intake with single tail vein streptozotocin(STZ)and four weeks of chronic unpredictable mild stress(CUMS).All drug administration groups were treated by gavage during CUMS modeling,and the control and model groups were given equal amount of distilled water.After four weeks,the serum levels of blood glucose and glycosylated hemoglobin were measured to determine the hypoglycemic effect of ZGJTJYF.Moreover,the open field test and Morris water maze test were performed to evaluate the antidepressant effect of ZGJTJYF.Changes in 5-hydroxytryptamine(5-HT)level were detected via high-performance liquid chromatography with electrochemical detection(HPLC-ECD);the levels of tryptophan(TRP),kynurenine(KYN),and indoleamine 2,3-dioxygenase(IDO)in the hippocampus were detected using enzyme-linked immunosorbent assay(ELISA);the protein expression levels of synaptophysin(SYN)and postsynaptic density material-95(PSD-95)were detected via immunohistochemistry(IHC);and the protein expression levels of N-methyl-D-aspartate receptor(NR)2 A and NR2 B were detected using Western blot.(ii)In vitro experiments:five SPF grade SD pregnant rats(E16–18)were used to obtain primary hippocampal neurons(Ne),six SD new-born rats were used to collected primary astrocytes(As)and microglia(MG),and to establish a Ne-As-MG co-culture system.All co-culture systems were divided into six groups:control(PBS),model[150 mmol/L glucose+200μmol/L corticosterone(G&P)+PBS],blank(G&P+blank serum),positive(G&P+positive drug-containing serum),ZGJTJYF(G&P+ZGJTJYF serum),and 1-methyl-D-tryptophan(1-MT,IDO inhibitor)(G&P+1-MT)groups.After 18 h of intervention by corresponding treatment,immunofluorescence was used to analyze the protein expression levels of SYN,PSD-95,NR2 A,and NR2 B;ELISA was performed to measure the levels of interleukin(IL)-1β,IL-6,tumor necrosis factor(TNF)-α,and TRP/KYN metabolic pathway-related factors[TRP,KYN,kynurenine acid(KYNA),quinolinic acid(QUIN)].Results(i)In vivo experimental results showed that ZGJTJYF-M and ZGJTJYF-L significantly improved the elevated blood glucose state of DD rats(P<0.01 and P<0.05,respectively);ZGJTJYF-H,ZGJTJYF-M,and ZGJTJYF-L increased their autonomous activity,learning,and memory ability(P<0.01,P<0.01,and P<0.05,respectively).Moreover,the levels of 5-HT and TRP were significantly increased(P<0.01),and the levels of KYN and IDO were significantly decreased in the hippocampus(P<0.01)of rats after ZGJTJYF-M treatment.The protein expression levels of SYN and PSD-95 were significantly upregulated in hippocampal neurons(P<0.01),while the abnormal activation of NR2A and NR2B was markedly inhibited in hippocampus(P<0.05)of rats after ZGJTJYF-M treatment.(ii)In vitro experimental results showed that ZGJTJYF-containing serum significantly increased the protein expression levels of SYN and PSD-95 in hippocampal neurons(P<0.01),decreased the levels of IL-1β(P<0.01),IL-6(P<0.05),TNF-α(P<0.01),IDO(P<0.05),KYN(P<0.05),and QUIN(P<0.01),and increased the levels of TRP and KYNA(P<0.01)in the simulated DD state.ZGJTJYF also had an significantly inhibitory effect on the abnormal activation of NR2A and NR2B in neurons(P<0.05)in a stimulated DD state.Conclusion ZGJTJYF can effectively improve 5-HT deficiency in the hippocampus of rats by inhibiting IDO expression and regulating the TRP/KYN metabolic pathway,and it has a favorable protective effect on hippocampal neuron injury caused by DD.Therefore,ZGJTJYF is an effective potential therapeutic drug for the prevention and treatment of DD.

Study on GC-MS fingerprint of petroleum ether fraction of Shenqi Jiangtang Granules

Objective To establish gas chromatography-mass spectrometry(GC-MS)fingerprint method for the petroleum ether fraction of Shenqi Jiangtang Granules(SQJTG)and evaluate the product quality.Methods The GC-MS fingerprint of petroleum ether fraction of SQJTG was established by GC-MS,and the chemical components corresponding to the fingerprint peaks were structurally identified on NIST2014.The batch consistency of SQJTG products was evaluated based on the chemical composition of petroleum ether parts by using fingerprint similarity evaluation and Principal components analysis(PCA)technology.At the same time,Hotelling's T2 and DMODX statistics are used to set the control range for the quality of different batches of products.Results Twenty-two components were identified from the petroleum ether part of SQJTG,accounting for 60.94%of the total components separated.The similarity of fingerprints of petroleum ether parts of 24 batches of SQJTG was greater than 0.95.The PCA of 24 batches of samples were all under the control limits of Hotellin’s T2 and DMODX statistics,indicating that the petroleum ether parts of different batches of SQJTG were consistent.Conclusion The developed GC-MS fingerprint method can be used to evaluate the quality of SQJTG.

Effects of Jiangtang Fanglong Wan on hearing in an animal model of diabetes

Objective ; To study effects of Jiangtang Fanglong Wan （glucose-lowering and deafness-preventing capsule） on hearing in an animal model of diabetes. Methods Wistar rats were used to create a diabetes model by intraperitoneal injection of streptozotocin （STZ, 55 mg/kg）. Forty rats were randomly selected to receive Jiangtang Fanglong Wan （ 10 g/kg/ day） through intragastric garage （treatment group） or normal saline （control group）. Auditory brainstem responses （ABRs） were recorded at Months 1, 2 and 3. Results ABR latencies and wave intervals were similar between the two groups at Month 1 （P 〉 0.05）. ABR latencies and wave intervals were shorter in the treatment group than those of the control group at Months 2 and 3 （P 〈 0.05 and P 〈 0.01, respectively）. Conclusion Our results suggest that Jiangtang Fanglong Wan may have a beneficial effect in preventing and treating hearing impairment associated with diabetes.

Integration of network pharmacology with experimental verification reveals the hypoglycemic mechanism of coptisine in Jinqi Jiangtang tablets:inhibition of the FoxO1 signaling pathway and hepatic gluconeogenesis

Background:Jinqi Jiangtang tablets(JQJT)have been approved for the treatment of type 2 diabetes mellitus(T2DM)in China for many years.Exploring the effective substances and mechanisms of JQJT is important for its clinical application and further drug research and development.This study aimed to explore the chemical basis and mechanisms of JQJT in the treatment of T2DM.Methods:With network pharmacology,we screened substances in JQJT and their possible targets,then constructed the action network and enriched the biological functions and pathways associated with the active components,and identified the potential targets and mechanisms of JQJT in the treatment of T2DM.Based on the network pharmacology data,we explored the hypoglycemic mechanisms of coptisine in JQJT through western blot and quantitative real-time polymerase chain reaction.Results:Forty-three compounds with good pharmacokinetic properties were identified in JQJT,together with 146 potential biological targets.Among these potential targets,74 were associated with treatment of T2DM.A compound-target network of the 43 compounds against T2DM was constructed.Biological process and signal pathway enrichment analysis of the network highlighted the FoxO signaling pathway.Western blot and quantitative real-time polymerase chain reaction results showed that coptisine,but not epiberberine,significantly inhibited expression of key genes involved in hepatocyte gluconeogenesis by regulating the FoxO1 signaling pathway.Conclusion:Network pharmacology analysis and cell experiments showed that coptisine regulated glucose homeostasis by inhibiting the FoxO1 signaling pathway and hepatic gluconeogenesis,which may be one of the mechanisms of JQJT in the treatment of T2DM.

Metabolomic profiling reveals hypoglycemic effect of Jinqi Jiangtang tablet on type 2 diabetic rats

Background:Si-Miao Sun wrote about the Jinqi Jiangtang tablet(JQJT)that is derived from the traditional Chinese herbal medicine Huanglian(Coptis Chinensis Franch)pills in the Essential Recipes for Emergent Use Worth A Thousand Gold in C.E.652.The tablet is used to treat diabetes in China owing to its powerful hypoglycemic properties.However,little is known about the metabolic mechanisms underlying these properties.Methods:Ultra-high-performance liquid chromatography coupled with quadrupole-time-of-flight tandem mass spectrometry-based metabolomics approach was performed to explore the metabolic mechanism of JQJT in the treatment of type 2 diabetes mellitus(T2DM).Results:The metabolomic pathway analysis and the Kyoto Encyclopedia of Genes and Genomes database were used to screen out 25 potential biomarkers and construct pertinent metabolic pathways.Five metabolites,such as allantoic acid and taurine,were downregulated and 20 metabolites were upregulated in the urine of T2DM rats.The biomarkers in the JQJT group exhibited a good callback trend.It is speculated that JQJT can alleviate T2DM by regulating the disorders of taurine,hypotaurine,phenylalanine,ascorbate,and aldarate metabolism,as well as pentose and glucuronate interconversions.Conclusion:This study will be meaningful in the clinical application of JQJT and will be valuable for further exploration of the metabolic mechanisms underlying its properties.

Effect of Jiangtang Bushen Recipe (降糖补肾方) in Intervention Treatment of Patients with Impaired Glucose Tolerance

Objective:To evaluate the intervention effect of diet, exercise and Jiangtang Bushen Recipe (JBR, 降糖补肾方), a Chinese herbal recipe, in preventing the progress of patients with impaired glucose tolerance ( IGT ) to diabetes mellitus (DM) type 2. Methods: Fifty-one IGT patients with their diagnosis conformed to the diagnosis standard of WHO, 1999, were randomly divided into the control group (n = 26) and the TCM group (n=25). Patients in the control group attended to the educational course for DM and received dietotherapy and kinetotherapy, and to those in the TCM group, under these treatments, JBR was given additionally. Oral glucose tolerance test (OGTT), body weight index (BWI), levels of blood lipids and fasting insulin of all the patients were examined after 3 months, 6 months and 12 months of treatment. The total observation time was 1 year. Results:Except the 6 cases out of the 51 patients (11.7%), on whom the observa-tion discontinued, in the control group, as compared with before treatment, levels of fasting insulin and fasting blood glucose after treatment were not changed significantly ( P > 0. 05 ) , also insignificant difference was shown in levels of total cholesterol (TC) and triglyceride (TG), thoughthe two indexes lowered slightly after treatment (P>0. 05), but significant difference was shown in comparison of OGTT/2h, blood glucose and BWI (P<0. 05). While in the TCM group, fasting blood glucose was changed insignificantly (P> 0. 05) , but there was significant difference in comparison of fasting insulin, TC, BWI, OGTT/2h and plasma glucose levels (P<0. 01) respectively before and after treatment. At the end of the stud-y, the cumulative cases with conversion to diabetes were 3 (13. 6%) in the control group, and 1 (4. 3%) in the TCM group, x2 test showed insignificant difference in comparison of diabetes conversion rate between the two groups (P>0. 05), however, the TCM group showed a better year conversion rate of normal glucose tolerance than that in the control group (x2 = 8. 31, P<0. 01). Conclusion: TCM intervention is possibly effective in delaying the conversion of IGT to DM type 2, and plays integrative effeciency in impelling IGT patients to health. The favorable education and treatment of DM controlling, including dieto- and kineto-therapy may also be advantageous in IGT intervention, but could not be effective in blocking the advance of IGT.Original article on CJITWM (Chin) 2004 ;24 (4): 317

Exploring the mechanisms of Jiangtang decoction in the treatment of diabetic nephropathy based on network pharmacology

Background:The purpose of this research is to predict the mechanisms of the experienced prescription Jiangtang decoction for treating diabetic nephropathy based on network pharmacology,the predicted targets and pathways were validated by celluar experiments.Methods:The active ingredients of the experienced prescription Jiangtang decoction and their putative targets were collected from TCMSP Database and SwissTargetPrediction platform.Diabetic nephropathy-related targets were excavated from GeneCards and DisGeNET database.Then,the interactions of potential targets of the experienced prescription Jiangtang decoction with well-known diabetic nephropathy targets were used to construct the protein-protein interaction network by STRING database and Cytoscape,and screened the core targets via topological analysis.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were performed by Metascape platform.Finally,we conducted in vitro experiments to verify this prediction of the network pharmacology.A diabetic nephropathy model was established by treating mesangial cells with D-ribose,in which the therapeutic effects of the experienced prescription Jiangtang decoction were evaluated.CCK-8 and LDH assay were used to test cell viability and cell toxicity,cell apoptosis was evaluated by Hoechst 33258 staining,AO/EB staining,levels of ROS were detected by fluorescent probe,the expression levels of MAPK signaling pathway-associated proteins and apoptosis-related proteins Bax were measured by western blotting assay.Results:A total of 74 active ingredients contained and 871 potential identified targets were retrieved from databases.Simultaneously,803 diabetic nephropathy-associated targets were also obtained,180 overlapped targets were considered as potential therapeutic targets of the experienced prescription Jiangtang decoction against diabetic nephropathy.By constructing a protein-protein interaction network and topological analysis,57 core targets were screened.Gene Ontology analysis highlighted 1655 Gene Ontology terms main including apoptotic signaling pathway,regulation of reactive oxygen species metabolic process and positive regulation of cell migration.KEGG enrichment analysis revealed that core targets were enriched mainly in MAPK,AGE-RAGE,TNF,PI3K-Akt signaling pathways.Cellular experiments further demonstrated D-ribose decreased mesangial cells viability,increased LDH release and the ROS level,induced apoptosis and activated the p38/JNK MAPK signal pathways,while the experienced prescription Jiangtang decoction could be useful in attenuation of D-ribose-induced damage.Conclusion:Network pharmacology intuitively shows the multi-component,multi-target and multi-pathway therapeutic effects of the experienced prescription Jiangtang decoction on diabetic nephropathy.By in vitro experiment,it revealed that the experienced prescription Jiangtang decoction has potential therapeutic effects on diabetic nephropathy via alleviating oxidative stress and apoptosis.the experienced prescription Jiangtang decoction treatment significantly inhibited the D-ribose-stimulated JNK and p38 MAPK activation.

The Mechanism of Action of Qihuang Jiangtang Capsule in the Treatment of Type 2 Diabetes Based on Network Pharmacology and Molecular Docking Technology

Objective Our objective was to investigate the potential mechanism of action of Qihuang Jiangtang capsule(QHJTC)in the treatment of type 2 diabetes mellitus(T2DM)through network pharmacology and molecular docking.Methods The active components of materia medica in the formula of QHJTC were searched on the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and Encyclopedia of Traditional Chinese Medicine.The targets related to the active components were obtained via PubChem database.The targets related to T2DM were retrieved through the GeneCards database.The targets corresponding to the active components and diabetes mellitus were uploaded to the Venn diagrams website to get the Venn diagram,and the intersecting targets were the potential targets of QHJTC in treating T2DM.The active components and potential targets were imported into Cytoscape 3.7.2 software to construct the active component–potential target network,and the key compounds and targets were screened by the Network Analyzer module in the Tools module.The potential targets were imported into the STRING database to obtain the interaction relationships,so as to analyze and construct the protein–protein interaction(PPI)network by Cytoscape 3.7.2 software.The intersecting targets were introduced into Metascape for gene ontology(GO)functional enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis.The top 20 signaling pathways obtained by the KEGG pathway enrichment analysis and the related targets and the corresponding targets were analyzed by using Cytoscape 3.7.2 software to construct the“active component–important target-key pathway network”for the intervention of T2DM with QHJTC.The molecular docking of active components and core targets was performed with AutoDock software.Results A total of 237 active components and 281 related targets were obtained from QHJTC,as well as 1362 T2DM targets and 155 potential targets of QHJTC in treating T2DM.There were 32 key components and 49 key targets identified by the active component–potential target network topology analysis.There were 471 terms obtained from GO functional enrichment analysis,among which 248 related to biological processes,125 related to molecular functions,and 98 related to cellular components.There were 299 signaling pathways obtained from KEGG pathway enrichment analysis.The active components of QHJTC were found spontaneously binding to the core targets.Conclusions QHJTC can treat T2DM through multi-components,multi-targets,and multi-pathways.

清血消脂降糖方对2型糖尿病大鼠胰岛素抵抗的影响及机制

目的探讨清血消脂降糖方对2型糖尿病(T2DM)大鼠胰岛素抵抗(IR)的改善作用及潜在机制。方法采用腹腔注射30 mg/kg链脲佐菌素联合高脂高糖饲料喂养的方法建立T2DM大鼠模型。实验设置正常对照组,模型组,清血消脂降糖方低、高剂量组(6.525、13.05 g/kg,以生药量计)和二甲双胍组(阳性对照,0.18 g/kg),每组8只。给药组大鼠灌胃相应药物,正常对照组、模型组大鼠灌胃等体积生理盐水,每日1次,持续6周。测定大鼠体重、空腹血糖(FBG)并进行口服葡萄糖耐量试验,检测大鼠血清中空腹胰岛素(FINS)水平并计算胰岛素抵抗指数(HOMA-IR)、胰岛素敏感指数(ISI),检测大鼠血清中血脂四项、肝功能、氧化应激指标及炎症因子水平,观察大鼠肝组织病理变化,检测大鼠肝组织中蛋白激酶R样内质网激酶(PERK)、叉头框蛋白O1(FOXO1)蛋白磷酸化水平。结果与模型组相比,清血消脂降糖方高剂量组和二甲双胍组大鼠体重、ISI、高密度脂蛋白胆固醇、超氧化物歧化酶水平显著升高(P<0.05),FBG、糖负荷120 min时的血糖值、葡萄糖曲线下面积、FINS、HOMA-IR、低密度脂蛋白胆固醇、总胆固醇、甘油三酯、丙氨酸转氨酶、天冬氨酸转氨酶、碱性磷酸酶、丙二醛、白细胞介素6、肿瘤坏死因子α、C反应蛋白水平均显著降低(P<0.05),肝组织病理损伤明显改善,肝组织中PERK、FOXO1蛋白磷酸化水平显著降低(P<0.05)。结论清血消脂降糖方可调节糖脂代谢、炎症因子及氧化应激水平,缓解T2DM大鼠IR,其作用机制可能与抑制PERK/FOXO1信号通路有关。

Chemical profiling of Jinqi Jiangtang tablets by HPLC-ESI-Q-TOF/MS

AIM： To profile the chemical constituents in Jinqi Jiangtang tablets. METHOD： Based on the chromatographic retention behavior, fragmentation patterns of chemical components, and published lit- eratures, a high-performance liquid chromatography coupled with electrospray ionization quadrnpole time-of-flight tandem mass spectrometry （HPLC-ESI-Q-TOF/MS） method was established to characterize and identify components in Jinqi Jiangtang tablets. RESULTS： A total of 52 chemical compounds, including eight iridoid glycosides, seven phenolic acids, twelve alkaloids, six fla- vonoids, and nineteen saponins, were identified in Jinqi Jiangtang tablets. CONCLUSION： The established method could serve as a powerful tool for structural characterization and quality control of this Chinese herbal preparation.

Jiangtang Xiaozhi Recipe(降糖消脂方)Prevents Diabetic Retinopathy in Streptozotocin-Induced Diabetic Rats

Objective： To evaluate the prevention effect of diabetic retinopathy of Jiangtang Xiaozhi Recipe （降糖消脂方, JXR） in streptozotocin （STZ）-induced diabetic rats. Methods： Sprague-Dawley rats were randomly divided into normal control group and diabetic group. Rats in the diabetic group were induced by intraperitoneal administration of STZ （50 mg/kg）, and subdivided into 5 groups. Rats in the diabetic control group were given saline; four treatment groups were given metformin （300 mg/kg）, JXR （2, 4 and 8 g/kg） respectively for 8 weeks, while rats in the normal control group were injected with citrate buffer and given the same volume of vehicle. Body weight and food intake were measured every week. The hypoglycaemic effects were determined by testing fasting blood glucose （FBG） every other week, and hemoglobin Alc （HbAlc）, insulin, and glucagon at the end of the treatment. The preventive effects of JXR on STZ-induced diabetic rats were determined by histopathological examination with hematoxylin and eosin staining, and periodic acid-schiff staining. The effects were further evaluated by serum superoxide dismutase （SOD） activity and malondialdehyde （MDA）. Results： High-dose JXR significantly reduced FBG and HbAlc level at the 8th week of administration （P〈0.01, P〈0.05）. JXR significantly increased insulin level （P〈0.05）, and decreased glucagon level （P〈0.05）. JXR showed the antioxidant defense with increased SOD activity and decreased MDA contents in diabetic rats. Histopathological studies revealed that there were no basement membrane thickening and mild destruction in the treated groups. Morphometric measurements of retina microvascular showed that acellular capillary and capillary density decreased in treated rats while pericyte and endothelial cell increasing after the treatment. Conclusion： JXR have protective effect of diabetic retinopathy and its mechanism may be associated with the obvious hypoglycemic and antioxidant effect.

Danzhi Jiangtang capsule(丹蛭降糖胶囊) alleviate hyperglycemiaand periodontitis via Wnt/β-catenin signaling in diabetic rats

OBJECTIVE:To elucidate whether Danzhi Jiangtang capsule(丹蛭降糖胶囊,DJC)has treatment effects on diabetic periodontitis and the potential mechanism.METHODS:One week after the induction of diabetes by streptozotocin(STZ),60 male Wistar rats were ligated by orthodontic ligation thread in cervical portion of bilateral maxillary first molar to induce diabetic periodontitis.Periodontitis was exanimated by tooth tissue morphology after 4 weeks.And then all rats were divided into 5 groups:diabetic periodontitis group(DP,n=20),periodontal basic treatment group(DP+BT,n=20),periodontal basic treatment+DJC treatment group(DP+BT+DJC,n=20)and additional Wnt/β-catenin inhibitor group(DP+BT+DJC+WIKI4/21 H7,n=20).Eight weeks after different interventions,fasting plasma glucose(FPG),C-peptide and glycosylated hemoglobin(Hb Alc)of rats were measured and then all rats were sacrificed.The paraffin sections of periodontal tissue were executed hematoxylin-eosin(HE)staining and immunohistochemical examination.The m RNA and protein levels of inflammatory cytokines were evaluated by quantitative polymerase chain reaction(Q-PCR)and Western blotting(WB).The protein levels of Wnt/β-catenin signaling molecules were measured by WB.RESULTS:The blood glucose and C-peptide concentrations in DP,DP+BT and DP+BT+DJC groups were gradually reduced,with gradually decreased distance of CEJ-A and the percentage of periodontal ligament(PDL),as well as gradually increased Hb Alc.The number of monocytes and leukocytes in the junctional epithelium and periodontal connective tissue was markedly decreased in DP+BT+DJC group(P<0.05),which was slightly reduced in DP+BT group comparing to DP group.The protein levels of Wnt1 andβ-catenin were obviously up-regulated with DJC treatment,while the SOST and DDK1 were markedly down-regulated with DJC treatment.The expression levels of BGP were lowest in DP group and highest in DP+BT+DJC group,while the tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ)and metalloproteinase-3(MMP-3)were highest in DP group and lowest in DP+BT+DJC group.All these changes could be reversed by Wnt/β-catenin inhibitor.CONCLUSION:DJC can improve the hyperglycemia and both distal alveolar bone loss and alveolar bone loss in furcation area of diabetic periodontitis rats by reducing the inflammation of gingival tissue and regulating the expressions of BGP,TNF-α,IFN-γand MMP-3 potentially through Wnt/β-catenin signaling.

Antioxidant capacity of extract from Jiangtang Xiaozhi recipe in vitro

OBJECTIVE: To evaluate the antioxidant capacity of the extract from Jiangtang Xiaozhi recipe(JXR) of in vitro.METHODS: JXR extract was prepared according to previously reported method. In vitro antioxidant assays were used in this experiment, including 1,1-Diphenyl-2-picrylhydrazyl free radical(DPPH) radical scavenging ability, 2-2’-Azinobis-(3-ethylbenzthiazoline-6-sul phonate(ABTS) radical scavenging ability, reducing power assay, fluorescence recovery after photo bleaching assay, β-carotene bleaching assay, ferric thiocyanate assay, and thiobarbituric acid method.RESULTS: DPPH, ABTS assay showed that JXR extract had distinct effect on scavenging free radicals;reducing power and ferricreducing-antioxidant power assay showed that JXR extract possessed redox ability;β-Carotene bleaching assay and antioxidant activity in a linoleic acid system using ferric thiocyanate method, thiobarbituric acid assay indicated that JXR extract could effectively inhibit lipid peroxidation, and the effect was better than that of Vitamin C.CONCLUSION: JXR extract has significant antioxidant capacity in vitro.

Jiangtang Xiaoke granule attenuates glucose metabolism disorder via regulating endoplasmic reticulum stress in the liver of type 2 diabetes mellitus mice

OBJECTIVE:To observe the effect of Jiangtang Xiaoke(JTXK) granule on endoplasmic reticulum(ER)stress in high fat diet(HFD)-induced type 2 diabetes mellitus(T2 DM) KK-Ay mice.METHODS:KK-Ay mice were fed with HFD to induce the T2 DM model,while normal control C57 BL/6 J mice were given standard feed.Fasting blood glucose(FBG) in all mice was measured weekly and oral glucose tolerance tests(OGTTs) were performed at 4 and 10 weeks after start of treatment to determine glucose metabolism.Serum fasting insulin(FINS) and insulin sensitivity index(ISI) were measured to determine insulin sensitivity.m RNA expressions of eukaryotic initiation factor-2 alpha(e IF2α),glucose regulated protein 78(GRP78),activating transcription factor 4(ATF4),and C/EBP homology protein(CHOP) were assessed by reverse transcription polymerase chain reaction and the protein expressions of p-e IF2α,GRP78,and CHOP were assessed by Western blotting.RESULTS:JTXK granule significantly reduced FBG and free fatty acid levels and improved OGTT at the120 min of the 10-week treatment in T2 DM KK-Ay mice.FINS and Hb Alc levels were reduced and insulin sensitivities were increased in KK-Ay diabetic mice,which were improved with the treatment of JTXK granule,especially at the 7 and 3.5 g/kg doses.JTXK granule at the 3.5 g/kg dose was most effective in reducing both gene and protein expressions of e IF2α,GRP78,and CHOP.CONCLUSION:ER stress response is increased in T2 DM KK-Ay mice.Treatment with JTXK granule attenuates glucose disorders,improves insulin sensitivity,and reduces serum FFA in T2 DM KK-Ay mice.The mechanisms may be attributed to regulation of the signaling ER stress pathway via decreasing e IF2α phosphorylation and suppressing e IF2α-ATF4-CHOP activation.

Qizhi Jiangtang capsule(芪蛭降糖胶囊)activates podocyte autophagy in diabetic kidney disease by inhibiting phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin pathways

OBJECTIVE:To investigate the therapeutic action and mechanism of the Qizhi Jiangtang capsule(芪蛭降糖胶囊,QZJT)on diabetic kidney disease(DKD)treatment.METHODS:This experiment used db/db mice and podocytes(MPC5)to develop DKD model.Evaluation of the effect of the QZJT on db/db mice by testing urine and blood biochemical parameters(24-h urinary albumin,serum creatinine,blood urine nitrogen),pathological kidney injury,and podocyte integrity.Moreover,autophagosomes in podocytes of DKD mice and cultured podocytes were detected using electron microscopy.Additionally,Western blotting was applied to detect the expression of podocyte marker protein(podocin),autophagy-associated proteins,and phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/AKT/mTOR)signaling pathway changes in vivo and in vitro.RESULTS:QZJT significantly reduced urine protein,blood nitrogen urea,and serum creatinine and showed histological restoration of renal tissues.QZJT also significantly improved the down-regulation of podocin and foot fusion and effacement in db/db mice.QZJT increased autophagic vesicles in mice and cultured podocytes.QZJT also upregulated microtubuleassociated protein 1 light chain 3-II(LC3-II)/(LC3-I)and Beclin-1 and downregulated phosphorylated-PI3K(pPI3K),p-AKT,and p-mTOR in db/db mice and MPC5 cells.However,autophagy inhibitor 3-methyladenine partially alleviated the above effects in MPC5 cells.CONCLUSIONS:These results showed that the QZJT can enhance podocyte autophagy and ameliorate podocyte injury in DKD by inhibiting the PI3K/AKT/mTOR signaling pathway.

Danzhi Jiangtang capsule(丹蛭降糖胶囊)reduces renal injury in rats with diabetes induced by high fat diet and streptozotocin via downregulating toll-like receptor 4-nuclear factor-κB pathway and apoptosis

OBJECTIVE:To investigate the effect and mechanisms of Danzhi Jiangtang capsule(丹蛭降糖胶囊,DJC)on renal injury in streptozotocin(STZ)-induced diabetes of rats.METHODS:Sprague-Dawley rats were fed with high fat diet for 6 weeks followed by streptozotocin(STZ,35 mg/kg)injection.These rats were then treated with DJC(270,540 and 1080 mg/kg)daily for 8 weeks.RESULTS:A combination of high fat diet and STZ significantly increased blood glucose creatinine,urea nitrogen,and urine albumin in rats.Meanwhile,the glomerular and tubular lesions were observed in rats fed with high fat diet and injected with STZ.These biochemical and pathological changes were significantly attenuated by DJC treatments in a dose-dependent manner.Mechanistically,DJC treatments significantly decreased toll-like receptor 4(TLR4),mitogen-activated protein kinase(MAPK),and nuclear factor-κB(NF-κB)signals in the kidney of rats fed with high fat diet and injected with STZ.Terminal deoxynucleotidyl transferase dU TP nick end labeling staining and caspase-8 levels showed that renal apoptosis was increased in rats fed with high fat diet and injected with STZ,and this was attenuated by DJC treatments.CONCLUSIONS:DJC treatments protect against diabetic kidney disease,and the mechanism may be closely related to downregulation of TLR4/MAPK/NF-κB pathways and apoptosis.This study provides further evidence of using DJC as a potential therapeutic option for diabetic kidney disease.

葛芪降糖汤治疗气阴两虚型2型糖尿病临床观察

目的:比较研究在常规西药治疗的基础上,观察葛芪降糖汤对气阴两虚型2型糖尿病的临床疗效。方法:2023年7月—2024年7月治疗的60例2型糖尿病患者,随机分为治疗组和对照组各30例。其中对照组使用常规西药方案治疗,治疗组在对照组的治疗方案基础上加入葛芪降糖汤配合治疗3个月,观察治疗前后患者血糖、糖化血红蛋白和血脂的变化以及不良反应的出现情况,进行两组患者临床疗效比较。结果:治疗组临床有效性明显,能更有效地降低患者空腹和餐后血糖水平、降低糖化血红蛋白和血脂水平(P<0.05)。治疗组在不良反应发生率方面也显著低于对照组(P<0.05)。结论:配合葛芪降糖汤治疗2型糖尿病(气阴两虚型),效果明显,且不良反应少,值得今后的临床工作中推广应用。

Effect of Sanhuang Jiangtang (三黄降糖) Recipe on Insulin Peripheral Resistance in Type Ⅱ Diabetes Mellitus

Objective: To observe the effect of Sanhuang Jiangtang recipe (三黄降糖, SAT) on insulinperipheral resistance in Type Ⅱdiabetes mellitus (DM). Methods:Ninety-five patients with type Ⅱ DM wererandomly divided into two groups. Fifty-three cases of SHJT group were given decoction and tablets of SAT orally for 4 -- 6 months. The efficacy was compared with that of 42 cases treated with Glipizide as the control.Before and after treatment standard steamed bread meal test was performed to measure the insulin peripheralsensitivity, insulin release to glucose and insulin sensitivity index. Results: (1 ) The total effective rates of improving insulin peripheral resistance and reducing blood sugar in SHJT group were 79. 2 % and 80. I %, whichwas equivalent to levels in the control group, but SAT recipe was more effective in relieving symptoms of Qideficiency and signs of blood stasis. (2) In SalT group, the insulin peripheral sensitivity and insulin sensitivityindex were significantly increased (P < 0. 05 and 0. of ), meanwhile the fasting blood sugar and blood sugar areawere reduced (P < 0. 05), but the change of insulin release to glucose was blunted. (3 )The lowering of bloodsugar in SHJT group was significantly negative correlated with the changing of degree of insulin peripheral sensitivity and index of insulin sensitivity (P < 0. 01 and 0. 05), but not with that of insulin area. Conclusion: Itsuggested that the treatment of SHJT recipe might decrease insulin peripheral resistance (partial reversal) bymeans of reducing hyperinsulinemia and improving insulin sensitivity.