Clinical Effect of Modified Gegen Qinlian Decoction Combined with Western Medicine on Children with Rotavirus Enteritis with Damp-Heat Syndrome

[Objectives] To explore the clinical effect of Modified Gegen Qinlian Decoction combined with western medicine on children with rotavirus enteritis with damp-heat syndrome and its influence on myocardial enzymes.

Mechanism of Gegen Qinlian Decoction in Treating Type 2 Diabetes Mellitus Complicated with NAFLD Based on Network Pharmacology

[Objectives]To explore the mechanism of Gegen Qinlian Decoction in treating type 2 diabetes mellitus(T2DM)complicated with non-alcoholic fatty liver disease(NAFLD)by analyzing the effective components of Gegen Qinlian Decoction.[Methods]TCMSP database was used to analyze the active components of Gegen Qinlian Decoction,and pubchem and Swiss ADME databases were also used to predict drug targets,extract T2DM complicated with NAFLD targets from OMIM and Genecards databases.Venny plot was drawn to obtain intersection targets,and finally Cytoscape was used to make core target maps and drug-target-disease network maps.Using DAVID and Metascape database to analyze the intersection targets,the gene ontology information of Go and KEGG was obtained.Microbial informatics technology was used to visualize GO,and Cytoscape was used to make drug-target-disease network map-enrichment pathway map.[Results]The network pharmacological analysis showed that Gegen Qinlian Decoction acted on the key targets of type 2 diabetes mellitus complicated with non-alcoholic fatty liver disease,such as ALB and ALT1,through many components,and achieved the purpose of treating this disease.The chemical constituents of the drug include formononetin,5-hydroxyisomucronulatol-2,5-2-O-glucoside,cholesteryl laurate,isoliquiritigenin,etc.[Conclusions]This study provides a new idea and theoretical support for future drug research and clinical practice.

The potential role of Gegen Qinlian decoction in the treatment of coronavirus disease 2019 based on network pharmacology and validation of lipopolysaccharide-induced macrophages

Background:Coronavirus disease 2019(COVID-19)is caused by severe acute respiratory syndrome coronavirus 2,which has led to deaths and currently lacks an efficient treatment.Despite studies suggesting the potential of the Gegen Qinlian decoction(GQD)in preventing COVID-19,comprehensive analyses of its anti-COVID-19 potential are still lacking.Methods:GQD treatment was evaluated for its efficacy in ameliorating the early stage(24 hours)of lipopolysaccharide(LPS)-induced cytokine storm in vivo.Additionally,target genes of GQD were co-analyzed with COVID-19 signature genes to identify key ingredients and their pathways.Validation was also conducted using an LPS-induced macrophage model.Results:GQD treatment effectively ameliorated the early stage of LPS-induced cytokine storm in vivo.Key ingredients such as quercetin were found to be involved in multiple pathways,including inflammation,immunity,oxidative stress,cell proliferation,and apoptosis,through the AGE-RAGE signaling pathway and IL-17 signaling pathway.In the LPS-induced macrophage model,quercetin inhibited macrophage polarization(M1)and the secretion of inflammatory factors(IL-6,TNF-α,IL-17A).Conclusions:Our results indicate that GQD can be utilized in the treatment of cytokine storm induced by COVID-19 and has the potential to treat COVID-19 by suppressing the COVID-19 signature genes and macrophage polarization.

Gegen Qinlian decoction enhances immunity and protects intestinal barrier function in colorectal cancer patients via gut microbiota

BACKGROUND We previously showed,using the Traditional Chinese Medicine System Pharmacology Database,that Gegen Qinlian decoction(GQD)had a direct antitumor effect,and was combined with programmed cell death protein(PD)-1 inhibitors to treat microsatellite stable(MSS)tumor-bearing mice.However,the effect of GQD on patients with colorectal cancer(CRC)is not clear.AIM To determine the therapeutic mechanism of GQD in improving immune function,reducing inflammation and protecting intestinal barrier function.METHODS Seventy patients with CRC were included in this study:37 in the control group and 33 in the treatment group.The proportions of CD4+T,CD8+T,natural killer(NK),NKT and T regulatory cells were measured by flow cytometry.Levels of the cytokines tumor necrosis factor(TNF)-α,interferon(IFN)-γ,interleukin(IL)-2,IL-6,IL-10 and serotonin(5-hydroxytryptamine;5-HT)in serum were assessed by enzyme-linked immunosorbent assay(ELISA).The expression of zonula occludens(ZO)-1,occludin,nuclear factor(NF)-κB and TNF-αin tumor and normal tissues was measured by immunohistochemistry.The composition of gut microbiota from patients in the treatment group was assessed using 16S rDNA analysis.RESULTS There were no adverse events in the treatment group.The proportion of CD4+T cells and NKT cells in the post-treatment group was significantly higher than that in the pre-treatment and control groups(P<0.05).The level of TNF-αin the posttreatment group was significantly lower than that in the pre-treatment and control groups(P<0.05).The concentration of 5-HT in the post-treatment group was significantly lower than that in the pre-treatment group(P<0.05).The expression of ZO-1 and occludin in tumor tissues in the treatment group was significantly higher than that in the control group(P<0.05).The expression of ZO-1 in normal tissues of the treatment group was significantly higher than that in the control group(P=0.010).Compared with the control group,expression of NF-κB and TNF-αin tumor tissues of the treatment group was significantly decreased(P<0.05).Compared with the pre-treatment group,GQD decreased the relative abundance of Megamonas and Veillonella.In addition,GQD increased the relative abundance of Bacteroides,Akkermansia and Prevotella.CONCLUSION GQD enhances immunity and protects intestinal barrier function in patients with CRC by regulating the composition of gut microbiota.

LC-ESI-MS/MS,a modified method for simultaneous quantification of isoflavonoids,flavonoids,alkaloids and saponins in a Chinese herbal preparation Gegen-Qinlian decoction

A sensitive and reliable liquid chromatography-electrospray ionisation-tandem mass spectrometry(LC-ESI-MS/MS)method was established to simultaneously quantitate four categories of compounds(isoflavonoids,flavonoids,alkaloids and saponins)in Gegen-Qinlian decoction(GQD).These compounds were separated by a Shiseido CAPCELL PAK C18 column with a linear gradient consisting of 0.1%(v/v)formic acid in water(A)and 0.1%(v/v)formic acid in acetonitrile(B),and delivered at a flow rate of 0.3 mL/min.All the analytes were determined by electrospray positive ionization tandem mass spectrometry in a multiple reaction monitoring(MRM)mode.Linearity,accuracy,precision,recovery and stability of the method were evaluated with the validation over the range of 4.0-538 5 ng/mL.The proposed method was applied to the analysis of a Chinese herbal preparation GQD successfully.

Administration of modified Gegen Qinlian decoction for hemorrhagic chronic radiation proctitis: A case report and review of literature

BACKGROUND Hemorrhagic chronic radiation proctitis(CRP) is a common late complication of irradiation of the pelvis and seriously impairs life quality. There is no standard treatment for hemorrhagic CRP. Medical treatment, interventional treatment, and surgery are available, but they are limited in their applications due to nondefinite efficacy or side effects. Chinese herbal medicine(CHM), as a complementary or alternative therapy, may provide another option for hemorrhagic CRP treatment.CASE SUMMARY A 51-year-old woman with cervical cancer received intensity-modulated radiation therapy and brachytherapy with a total dose of 93 Gy fifteen days after hysterectomy and bilateral adnexectomy. She received six additional cycles of chemotherapy with carboplatin and paclitaxel. Nine months after radiotherapy treatment, she mainly complained of 5-6 times diarrhea daily and bloody purulent stools for over 10 d. After colonoscopy examinations, she was diagnosed with hemorrhagic CRP with a giant ulcer. After assessment, she received CHM treatment. The specific regimen was 150 mL of modified Gegen Qinlian decoction(GQD) used as a retention enema for 1 mo, followed by replacement with oral administration of 150 mL of modified GQD three times per day for 5 mo. After the whole treatment, her diarrhea reduced to 1-2 times a day. Her rectal tenesmus and mild pain in lower abdomen disappeared. Both colonoscopy and magnetic resonance imaging confirmed its significant improvement. During treatment,there were no side effects, such as liver and renal function damage.CONCLUSION Modified GQD may be another effective and safe option for hemorrhagic CRP patients with giant ulcers.

Study on the molecular mechanism of Gegen Qinlian decoction in the treatment of influenza based on network pharmacology

Objective:The study shows that Gegen Qinlian decoction(GGQL)is feasible in the treatment of influenza.It is explained from traditional Chinese medicine theory and modern pharmacology,and the formula is tested in vitro.The results show that Gegen Qinlian decoction has a good inhibitory effect on influenza virus.Methods:Search TCMSP database to get the components and targets of GGQL,and construct the compound target network.Then search OMIM database and Disgenet database to obtain influenza genes,and the common genes were obtained by VennDiagram.STRING database was used to construct gene function association network.Finally,David database was used to analyze GO enrichment and pathway enrichment.Results:146 ingredients and 128 targets were obtained,588 genes were obtained.46 common genes were obtained by VennDiagram,and import the common genes into the STRING database and construct the gene function association network,then the network information data was imported into Cytoscape software for topology analysis.Finally,13 enriched GO clusters and 7 enriched pathway clusters were obtained by David database,and construct the targets-GO-pathway network.Conclusion:Network pharmacology of GGQL reveal that the target and the pathway of the drug was closely related to influenza,and the results of this study have been verified with published experimental studies,which shows that GGQL has not only theoretical basis for the treatment of influenza,but also in vitro experiments support this conclusion.

Explanation of Gegen Qinlian Decoction Syndrome and Its Clinical Application

Gegen Qinlian Decoction is one of the commonly used classical prescriptions,which consists of fourherbs:Gegen(Puerariae Lobatae Radix),Huangqin(Scutellariae Radix),Huanglian(Coptidis Rhizoma),and Zhigancao Glycyrrhizae Radix et Rhizoma Praeparata cum Melle.The clinical application experience of Gegen Qinlian Decoction is as follows:first,according to Zhongjing Zhang's original text,heat distressing the large intestine with unresolved exterior syndrome is theoriginal meaningof Gegen Qinlian Decoction syndrome.In clinical practice,bacillary dysentery,gastrointestinal cold,etc.,belong to simultaneous exterior and interior disease,which are completely consistent with the original meaning of this prescription;second,the clinical digestive system diseases with exuberant heat of the large intestine as the core manifestation cannot be treated merely based on the exterior syndrome;third,due to the interior-exterior relationship between the lung and the large intestine and between the meridians,the use of Gegen(Puerariae Lobatae Radix),Huangqin(Scutellariae Radix),and Huanglian(Coptidis Rhizoma)can make the source of body clear,and the waste qi can be released and thepore can be easily opened,so this prescription can also be used for exogenous fever;fourth,with multidimensional comprehensive understanding of the syndrome,pathogenesis,symptoms and pharmacology,Gegen Qinlian Decoction has also been further applied to the treatment of hypertension,diabetes,etc.

Double-blinded,randomized clinical trial of Gegen Qinlian decoction pinpoints Faecalibacterium as key gut bacteria in alleviating hyperglycemia

Background:Accumulating evidence suggests that metabolic disorders,including type 2 diabetes mellitus(T2DM),can be treated with traditional Chinese medicine formulas,such as the Gegen Qinlian decoction(GQD).This study elucidates the mechanisms by which gut microbes mediate the anti-diabetic effects of GQD.Methods:We conducted a double-blind randomized clinical trial involving 120 untreated participants with T2DM.During the 12-week intervention,anthropometric measurements and diabetic traits were recorded every 4 weeks.Fecal microbiota and serum metabolites were measured before and after the intervention using 16S rDNA sequencing,liquid chromatography-mass spectrometry,and Bio-Plex panels.Results:Anti-diabetic effects were observed in the GQD group in the human trial.Specifically,glycated hemoglobin,fasting plasma glucose,and two-hour postprandial blood glucose levels were significantly lower in the GQD group than in the placebo group.Additionally,Faecalibacterium was significantly enriched in the GQD group,and the short-chain fatty acid levels were higher and the serum inflammation-associated marker levels were lower in the GQD group compared to the placebo group.Moreover,Faecalibacterium abundance negatively correlated with the levels of serum hemoglobin,fasting plasma glucose,and pro-inflammatory cytokines.Finally,the diabetes-alleviating effect of Faecalibacterium was confirmed by oral administration of Faecalibacterium prausnitzi(DSMZ 17677)in T2DMmousemodel.Conclusions:GQD improved type 2 diabetes primarily by modulating the abundance of Faecalibacterium in the gut microbiota,alleviating metabolic disorders and the inflammatory state.

Antidiabetic Effects of Gegen Qinlian Decoction via the Gut Microbiota Are Attributable to Its Key Ingredient Berberine

Gegen Qinlian Decoction(GQD),a traditional Chinese medicine(TCM)formula,has long been used for the treatment of common metabolic diseases,including type 2 diabetes mellitus.However,the main limitation of its wider application is ingredient complexity of this formula.Thus,it is critically important to identify the major active ingredients of GQD and to illustrate mechanisms underlying its action.Here,we compared the effects of GQD and berberine,a hypothetical key active pharmaceutical ingredient of GQD,on a diabetic rat model by comprehensive analyses of gut microbiota,short-chain fatty acids,proinflammatory cytokines,and ileum transcriptomics.Our results show that berberine and GQD had similar effects on lowering blood glucose levels,modulating gut microbiota,inducing ileal gene expression,as well as relieving systemic and local inflammation.As expected,both berberine and GQD treatment significantly altered the overall gut microbiota structure and enriched many butyrate-producing bacteria,including Faecalibacterium and Roseburia,thereby attenuating intestinal inflammation and lowering glucose.Levels of short-chain fatty acids in rat feces were also significantly elevated after treatment with berberine or GQD.Moreover,concentration of serum proinflammatory cytokines and expression of immune-related genes,including Nfkb1,Stat1,and Ifnrg1,in pancreatic islets were significantly reduced after treatment.Our study demonstrates that the main effects of GQD can be attributed to berberine via modulating gut microbiota.The strategy employed would facilitate further standardization and widespread application of TCM in many diseases.

Effect of Puerarin on the Pharmacokinetics of Baicalin in Gegen Qinlian Decoction (葛根芩连汤) in Mice

Objective： To study the pharmacokinetics of puerarin（PUE） in Gegen Qinlian Decoction（葛根芩连汤, GQD）, and the effects of PUE dosage variations on the pharmacokinetics of baicalin（BAL） in mice. Methods： GQD is composed of the concentrated granules of four Chinese herbs. Three dosages with different levels of PUE, including GQD, GQD co-administered with PUE, and GQD co-administration with two times the amount of PUE, were used to research the pharmacokinetics of PUE and BAL in mice. The indirect competitive enzyme-linked immunosorbent assay（ic ELISA） methods based on an anti PUE-monoclonal antibody（MAb） and BAL-MAb were employed to determine the concentration of PUE and BAL in mice blood. Results： After the co-administration of GQD with PUE, the area under the curves（AUC0-14 h） of PUE increased 2.8 times compared with GQD. At the dose of GQD co-administration at two times that of PUE, the AUC0-14 h of PUE was almost equal to that of GQD co-administration of PUE, showing non-linear pharmacokinetics. The AUC0-48 h of BAL showed a good dose-related increase of PUE（r=0.993） in the range from 100 to 300 mg/kg, indicating that PUE dramatically affects the absorption of BAL in mice. There was no significant difference in the other pharmacokinetic parameters, such as the first time of maximum concentration（Tmax）, the second Tmax, or the mean residence time. Conclusions： The ic ELISA methods were successfully applied to pharmacokinetic studies of PUE and BAL in GQD in mice. The dosage variability of PUE of the main ingredient in GQD affects its own pharmacokinetic characteristics and the absorption characteristics of BAL.

Comprehensive Metabolic Profiling of Modified Gegen Qinlian Decoction by Ultra-high-Performance Liquid Chromatography-Diode Array Detection-Q-Exactive-Orbitrap-Electrospray Ionization-Mass Spectrometry/Mass Spectrometry and Application of High-Performance

Objective:Gegen Qinlian decoction(GQD)is a classical traditional Chinese medicine formulation which has been used for almost 2000 years.At Guang'anmen Hospital,Beijing,a modified GQD version(m GQD)with seven instead of four herbal ingredients has been applied to treat Type 2 diabetes.Quality control is a crucial prerequisite for the therapeutic application of herbal medicines.For the identification of products derived from classical GQD,the Chinese Pharmacopeia requires the analysis of only three marker compounds.Because m GQD is a more complex mixture containing seven herbs and hundreds of constituents,the pharmacopoeia method for GQD is inadequate.Materials and Methods:A more comprehensive characterization of the formula's constituents has been developed using ultra-high-performance liquid chromatography-diode array detection(UHPLC-DAD)-Q-Exactive-mass spectrometry(MS)in electrospray ionization positive and negative mode.Moreover,a new method for the fingerprint analysis of m GQD via high-performance thin-layer chromatography(HPTLC)has been established.Results:Altogether,91 compounds have been assigned to their originating plants and 84 substances were identified either by comparison with authentic references or with data from the literature.The HPTLC method is based on the application of two different mobile phases and is able to detect both lipophilic and hydrophilic constituents of m GQD.Conclusions:The modified GQD was extensively characterized by UHPLC combined with DAD and Q-Exactive Orbitrap high-resolution MS detection,leading to the assignment and identification of compounds present in the decoction.In addition,a new method for the fingerprint analysis of the m GQD using HPTLC was established,which allows fast and simple identification of the herbal ingredients in the mixture.

葛根芩连汤加味治疗急性肠炎的临床观察

目的探究葛根芩连汤加味治疗急性肠炎的效果。方法纳入急性肠炎患者共80例,按照随机数字表法分为对照组与观察组各40例。对照组接受奥美拉唑等常规西医治疗,观察组在此基础上加用葛根芩连汤治疗。观察两组疗效,并比较治疗前后中医证候积分、炎症指标[肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL6)、超敏C反应蛋白(hs-CRP)、降钙素原(PCT)]、白蛋白、前白蛋白及免疫指标[免疫球蛋白G(IgG)、免疫球蛋白A(IgA)、免疫球蛋白M(IgM)]。结果观察组临床总有效率为95.00%,比对照组的80.00%高(P<0.05)。两组治疗后胃脘疼痛、身热不扬、大便稀溏、恶心呕吐评分比治疗前低,且观察组比对照组低(P<0.05);TNF-α、IL-6、hs-CRP及PCT比治疗前低,且观察组比对照组低(P<0.05);白蛋白、前白蛋白、IgG、IgA及IgM比治疗前高,且观察组比对照组高(P<0.05)。结论中西医结合治疗急性肠炎疗效确切,可明显抑制患者炎症反应,改善其血清白蛋白、前白蛋白水平。

葛根芩连汤加减联合二甲双胍对2型糖尿病胃肠湿热证患者中医证候积分及胰岛功能的影响

目的探讨葛根芩连汤加减联合二甲双胍对2型糖尿病(type 2 diabetes mellitus,T2DM)胃肠湿热证患者的影响。方法选取2022年4月—2023年12月深圳市中医院收治的78例T2DM胃肠湿热证患者为研究对象,根据不同治疗方法分为对照组和观察组,各39例。对照组口服盐酸二甲双胍片,观察组在对照组基础上给予葛根芩连汤加减治疗。对比两组的中医证候积分、血糖水平(空腹血糖、餐后2 h血糖、糖化血红蛋白)、胰岛功能(胰岛素抵抗指数、胰岛β细胞功能指数)、不良反应。结果治疗后,观察组的中医证候积分低于对照组,差异有统计学意义(P<0.05)。观察组空腹血糖、餐后2 h血糖、糖化血红蛋白水平均低于对照组,差异有统计学意义(P均<0.05)。治疗后,两组胰岛素抵抗指数水平均降低,且观察组低于对照组,两组胰岛β细胞功能指数均提高,且观察组高于对照组,差异有统计学意义(P均<0.05)。结论在T2DM胃肠湿热证患者中使用葛根芩连汤加减与二甲双胍联合治疗,可有效改善中医证候、血糖水平及胰岛功能,安全性高。

葛根芩连汤加减联合美沙拉嗪治疗大肠湿热型溃疡性结肠炎临床效果及其对肠道菌群、免疫功能和炎症水平的调节

目的观察葛根芩连汤加减联合美沙拉嗪治疗大肠湿热型溃疡性结肠炎的临床效果及其对患者肠道菌群、免疫功能和炎症水平的调节作用。方法选择2020年1月—2022年12月在医院消化内科治疗的100例溃疡性结肠炎患者作为研究对象,按照随机数字表法将患者平均分为美沙拉嗪组和葛根芩连组,各组50例,美沙拉嗪组患者单纯口服美沙拉嗪治疗,葛根芩连组患者在美沙拉嗪组基础上联合葛根芩连汤加减治疗。观察两组患者的临床效果和不良反应情况,并比较干预前后血清炎症因子、免疫功能以及肠道菌群变化。结果美沙拉嗪组患者的有效率为80.00%(40/50),葛根芩连组患者的有效率为94.00%(47/50),与美沙拉嗪组比较,葛根芩连组的有效率明显升高(P<0.05)。干预4周后,与美沙拉嗪组比较,葛根芩连组患者的血清白介素(IL)-6、IL-8、γ-干扰素(IFN-γ)以及肿瘤坏死因子-α(TNF-α)水平均明显下降(P<0.01);葛根芩连组患者外周血中CD_(3)^(+)、CD_(4)^(+)以及CD_(4)^(+)/CD_(8)^(+)水平明显升高,而CD_(8)^(+)水平则明显下降(P<0.01)。且葛根芩连组患者肠道双歧杆菌、乳酸杆菌以及肠球菌水平相较于美沙拉嗪组均明显升高,差异具有统计学意义(P<0.01)。干预4周后,葛根芩连组患者的IBDQ评分和SF-36评分相较于美沙拉嗪组均明显升高,差异具有统计学意义(P<0.01)。治疗期间两组患者的主要不良反应有腹胀、消化不良、头晕头痛、乏力、皮疹等,美沙拉嗪组和葛根芩连组的不良反应总发生率分别为10.00%(5/50)、14.00%(7/50),差异无统计学意义(P>0.05)。结论葛根芩连汤加减联合美沙拉嗪能够提高大肠湿热型溃疡性结肠炎患者的临床疗效,抑制血清炎症水平,调节机体免疫功能,其作用机制与调节患者肠道菌群平衡有关。

葛根芩连汤治疗溃疡性结肠炎研究进展

溃疡性结肠炎(ulcerative colitis,UC)是一种慢性复发性的结肠和直肠的炎症性疾病,其典型表现为腹痛、腹泻、黏液脓血便和吸收障碍等,属中医“肠澼”“痢疾”“滞下”“泄泻”“脏毒”等范畴。葛根芩连汤出自《伤寒论》,善清胃肠湿热,亦可调和气血,原方由葛根、黄芩、黄连和炙甘草4味中药组成,是治疗UC的经典良方。临床应用方面,主要通过葛根芩连汤加减、联合其他方剂、联合西药、联合灌肠、联合针灸等方式治疗UC患者,其优势体现在改善肠道黏膜屏障功能损伤,减少肠道炎症反应,缓解腹泻、腹痛、脓血便、里急后重、肛门灼热等症状,降低复发率,减少不良反应等,有效提高了UC患者的生活质量。作用机制方面,葛根芩连汤治疗UC主要与调节自身免疫、降低炎性细胞因子、调节T细胞、抗氧化应激、改善肠道菌群失调,调节IL-6/JAK2/STAT3、TLR4/NF-κB、Notch、Nrf2等信号通路有关。葛根芩连汤中单味药有效成分的研究表明,黄芩苷、黄芩多糖、葛根素、小檗碱、甘草酸二铵、甘草次酸等是治疗UC的主要有效成分。文章检索近年来国内外文献,从临床应用研究和实验机制研究两个方面综述葛根芩连汤治疗UC的研究进展,以期为葛根芩连汤治疗UC的遣方用药及后续研究葛根芩连汤治疗UC的作用机制提供参考依据,发挥中医药治疗UC的优势。

葛根芩连汤对2型糖尿病db/db小鼠胰腺组织PERK/ATF4/CHOP信号通路的影响

目的观察葛根芩连汤对2型糖尿病(T2DM)小鼠胰腺内质网应激的影响,探讨其治疗T2DM的作用机制。方法75只SPF级雄性db/db小鼠随机分为模型组、二甲双胍组和葛根芩连汤高、中、低剂量组,每组15只,15只db/m小鼠作为空白组,给药组分别予相应药物灌胃12周。检测小鼠体质量、空腹血糖(FBG)及糖化血红蛋白(HbA1c),HE染色观察胰腺组织病理变化,TUNEL染色检测胰岛细胞凋亡情况,Western blot检测胰腺组织葡萄糖调节蛋白78(GRP78)、蛋白激酶R样内质网激酶(PERK)、p-PERK、活化转录因子4(ATF4)、C/EBP同源蛋白(CHOP)蛋白表达,实时荧光定量PCR检测胰腺组织PERK、ATF4、CHOP mRNA表达。结果与空白组比较,模型组小鼠体质量、FBG和HbA1c含量显著增加(P<0.01);胰腺组织结构不完整,边界模糊,内有空泡,胰岛细胞凋亡明显增加(P<0.01);胰腺组织GRP78、p-PERK、ATF4、CHOP蛋白表达显著升高(P<0.01),PERK、ATF4、CHOP mRNA表达显著升高(P<0.01)。与模型组比较,各给药组小鼠体质量、FBG和HbA1c含量显著减少(P<0.05,P<0.01);胰腺组织病理改变有所减轻,胰岛细胞凋亡不同程度减少(P<0.05,P<0.01);葛根芩连汤高、中剂量组和二甲双胍组胰腺组织GRP78、p-PERK、ATF4、CHOP蛋白表达显著降低(P<0.01),PERK、ATF4、CHOP mRNA表达显著降低(P<0.05,P<0.01)。结论葛根芩连汤可能通过抑制内质网应激PERK/ATF4/CHOP信号通路,下调相关基因和蛋白表达,减少胰岛细胞凋亡,保护胰岛细胞功能,延缓T2DM进展。

葛根芩连汤对胰岛素抵抗大鼠肝脏能量代谢及游离脂肪酸的影响

目的探讨葛根芩连汤对胰岛素抵抗(IR)大鼠肝脏能量代谢及游离脂肪酸(FFA)的影响。方法采用60%脂肪的高脂饲料连续喂养13周复制IR大鼠模型。将SD大鼠随机分为正常组、模型组、罗格列酮组(5 mg·kg^(-1))以及葛根芩连汤低、中、高剂量组(1.65、4.96、14.86 g·kg^(-1)),每组6只。灌胃给药,每天1次,连续给药干预16周。测定IR相关指标:血清空腹胰岛素(FINS)、空腹血糖(FPG)值,计算IR指数;建立HPLC法测定大鼠肝组织中三磷酸腺苷(ATP)、二磷酸腺苷(ADP)、一磷酸腺苷(AMP)含量;采用ELISA法测定大鼠肝组织中的FFA含量;采用全自动生化分析仪检测血清总胆固醇(TC)、甘油三酯(TG)含量;采用HE染色法观察肝脏组织病理变化。结果(1)模型复制结束后,与正常组比较,模型组大鼠的FINS、FPG水平及IR指数显著升高(P<0.01);肝脏组织中ATP、ADP、AMP、FFA含量均无明显变化(P>0.05)。(2)给药干预结束后,与正常组比较,模型组大鼠的FINS、FPG水平及IR指数显著升高(P<0.01),肝脏组织中ATP、ADP、AMP含量均显著下降(P<0.01),FFA含量显著升高(P<0.01),血清TC、TG水平均明显升高(P<0.05);肝细胞排列紊乱,出现脂肪变性,并存在大量脂滴。与模型组比较,各给药组大鼠的FINS水平及IR指数均明显降低(P<0.05,P<0.01),罗格列酮组大鼠的FPG水平明显降低(P<0.05);各给药组大鼠肝脏组织中ADP、AMP含量均明显升高(P<0.05、P<0.01),葛根芩连汤低、中、高剂量组大鼠肝脏组织中ATP含量明显升高(P<0.05、P<0.01);罗格列酮组及葛根芩连汤低、高剂量组大鼠肝脏组织中FFA含量均显著降低(P<0.01);葛根芩连汤低、高剂量组大鼠血清TC水平明显降低(P<0.05,P<0.01),罗格列酮组及葛根芩连汤低剂量组大鼠血清TG水平明显降低(P<0.05,P<0.01);各给药组大鼠的肝细胞脂肪变性有不同程度减轻,且脂滴减少,病理损伤得到改善。结论葛根芩连汤可能通过调节能量代谢并降低FFA水平,从而改善肝脏脂质代谢紊乱,恢复脂质与能量平衡,从而起到改善IR的作用。

葛根芩连汤对肠道湿热证泄泻小鼠肠道机械屏障的影响

目的探讨葛根芩连汤对肠道湿热证泄泻模型小鼠肠道机械屏障的作用机制。方法选取15只SPF级雄性昆明小鼠,随机分为正常组5只和模型组10只。采取“高糖高脂+高温高湿+白酒稀释液+冰水法”复制肠道湿热证泄泻小鼠模型,造模成功的10只小鼠随机分为自愈组和治疗组,每组5只。治疗组灌胃5.07 g/(kg·d)葛根芩连汤水煎液,正常组和自愈组给予等容积无菌水,连续灌胃4 d后无菌提取各组小鼠十二指肠、空肠和回肠肠段。观察各组小鼠一般状态;HE染色观察各组肠段形态;采用Image J图像处理软件测量各组绒毛长度(villus height,VH)、隐窝深度(crypt depth,CD),计算VH与CD的比值(绒隐比,V/C)、杯状细胞数目和紧密连接蛋白平均光密度值(average optical density value,AOD);采用二步法免疫组织化学法分析各肠段密封蛋白1(Claudin 1)、闭合蛋白(Occludin)和紧密连接蛋白-1(zonula occluden-1,ZO-1)的表达。结果正常组小鼠毛发有光泽,精神状态佳,饮水量、肛温和二便均正常;治疗组小鼠毛发洁白、精神状态良好、大便颗粒饱满、肛温恢复正常、泄泻症状消失;自愈组小鼠毛发暗淡、肛温高、大便含水量较多、挤压不成型。与正常组相比,自愈组十二指肠、空肠和回肠肠黏膜结构部分受损,小肠腺出现萎缩,出现炎性症状。与自愈组相比,治疗组肠黏膜结构较清晰,肠上皮细胞和固有层细胞排列紧密,炎症情况明显改善。与正常组相比,自愈组十二指肠CD升高(P<0.01),十二指肠V/C和杯状细胞数降低(P<0.05,P<0.01);治疗组十二指肠V/C降低(P<0.05)。与正常组相比,自愈组空肠、回肠VH和V/C均明显降低(P<0.05,P<0.01)。与自愈组相比,治疗组十二指肠CD显著降低(P<0.01),十二指肠、回肠V/C升高(P<0.05,P<0.01)。十二指肠肠段,与正常组相比,自愈组和治疗组Claudin 1、Occludin、ZO-1 AOD值降低(P<0.05,P<0.01);与自愈组相比,治疗组Claudin 1、Occludin、ZO-1 AOD值显著升高(P<0.01)。与正常组比较,自愈组空肠、回肠肠段Occludin AOD值明显降低(P<0.01)。与自愈组比较,治疗组空肠肠段Claudin 1 AOD值升高(P<0.05),回肠肠段Occludin AOD值明显升高(P<0.01)。结论葛根芩连汤可改善肠道湿热证泄泻小鼠肠道炎症水平、促进紧密连接蛋白Claudin1、Occludin和ZO-1的表达,其机制可能与修复肠道机械屏障功能有关。

加减葛根芩连汤对感染耐药性大肠杆菌雏鸡的治疗效果

为了评价加减葛根芩连汤对感染耐药性大肠杆菌雏鸡的治疗效果,本试验首先用耐药性大肠杆菌感染4日龄雏鸡,6 h后分别灌服低(0.125 mL/羽)、中(0.25 mL/羽)和高(0.5 mL/羽)剂量的加减葛根芩连汤、锦心口服液(中药对照组)、恩诺沙星(西药对照组)和生理盐水(感染对照组和空白对照组),连续灌服5 d,计算各组雏鸡存活率和治愈率、采食量和饮水量、体重及其增量,分析血液生理生化指标和免疫器官指数。结果显示,低、中和高剂量组雏鸡24 h存活率分别为85%、80%和65%,极显著高于中药对照组(50%)和西药对照组(50%)(P<0.01);中和高剂量组雏鸡7d存活率和治愈率均达70%,极显著高于感染对照组和西药对照组(P<0.01);各感染组雏鸡每日平均采食量和平均饮水量在感染后明显减少,用药后均恢复并趋于稳定,体重保持稳定增长。血液生理生化指标结果显示,低、中和高剂量组白细胞数目显著低于西药对照组(P<0.05),平均血红蛋白含量显著高于空白对照组(P<0.01、P<0.001、P<0.01),中和高剂量组平均红细胞血红蛋白浓度显著低于空白对照组(P<0.05),高剂量组红细胞分布宽度变异系数显著高于其他各组(P<0.05),低、中和高剂量组肌酐含量极显著低于西药对照组(P<0.01)。免疫器官指数结果显示,低和中剂量组感染雏鸡法氏囊和脾脏指数极显著高于西药对照组(P<0.01),高剂量组胸腺指数显著高于中药对照组(P<0.05)。结果表明,不同剂量的加减葛根芩连汤对耐药性大肠杆菌感染雏鸡均具有较好的治疗效果,且中和高剂量的治疗效果显著优于低剂量,具有替代抗生素的潜力,可为耐药性大肠杆菌病的防治提供理论支撑。