Effects of Gegen Qinlian Decoction on NLRP3 inflammatome signaling pathway in liver of mice with type 2 diabetes mellitus / 国际中医中药杂志

Objective:To observe the effects of different doses of Gegen Qinlian Decoction on nucleotide oligomeric domain-like receptor protein 3 (NLRP3)/Caspase-1/IL1β inflammatory signaling pathway in liver of db/db mice with type 2 diabetes mellitus (T2DM).Methods:Totally 75 SPF male db/db mice were randomly divided into model group, metformin group (0.2 g/kg), Gegen Qinlian Decoction high-, medium-, and low-dosage groups (61.80, 30.90, 15.45 g/kg), with 15 mice in each group. Another 15 db/m male mice were selected as blank control group. Each administration group was given relevant medicine for gavage, while the blank group and model group were given 0.9% sodium chloride solution for gavage, once a day, for 12 weeks. The body weight, fasting blood glucose (FBG) and glycosylated hemoglobin (HbA1c) contents of each group were measured after treatment. The mRNA expression levels of NLRP3, Caspase-1 and interleukin-1β (IL-1β) in liver were detected by real-time quantitative PCR. The expression levels of NLRP3, Caspase-1, IL-1β and IL-18 in liver tissues were detected by Western Blot. HE staining was used to observe the morphology of liver tissues.Results:Compared with model group, body weight, fasting blood glucose and HbA1c contents of mice in Gegen Qinlian Decoction high- and medium-dosage groups and metformin groups decreased ( P<0.05), the body weight and fasting blood glucose levels of mice in Gegen Qinlian Decoction low-dosage group decreased ( P<0.05); the mRNA levels of NLRP3 and IL-1β in liver tissues of all treatment groups decreased ( P<0.05), the mRNA level of Caspase-1 in liver tissue decreased in Gegen Qinlian Decoction high- and medium-dosage groups ( P<0.05); the expression of NLRP3, Caspase-1, and IL-18 in liver tissue of each treatment group decreased ( P<0.05), while the expression of IL-1β in Gegen Qinlian Decoction high- and medium-dosage groups and the metformin group decreased ( P<0.05); compared with the metformin group, the body weight and fasting blood glucose of mice in the Gegen Qinlian Decoction high-dosage decreased ( P<0.05), while the HbA1c levels in the Qinlian Decoction high- and medium-dosage decreased ( P<0.05); the expressions of NLRP3, Caspase-1 and IL-18 in liver tissues of Gegen Qinlian Decoction high-dosage group decreased ( P<0.05), the expression of IL-1β, NLRP3, Caspase-1, IL-1β, and IL-18 decreased ( P<0.05); HE staining showed that the pathological changes of liver tissue were reduced in all treatment groups. Conclusion:Gegen Qinlian Decoction may reduce blood sugar by inhibiting the activation of NLRP3/Caspase-1/IL-1β inflammatory signaling pathway in liver of db/db mice, thereby improving the inflammatory damage of T2DM.

Mechanism of heat-clearing prescriptions in alleviating type 2 diabetes mellitus:a review / 中国中药杂志

Type 2 diabetes mellitus(T2DM), a common chronic metabolic disease, is often accompanied by internal heat syndrome. Heat-clearing prescriptions are widely used to treat different heat syndromes of T2DM from the aspects of clearing stagnant heat, excess heat, damp heat, phlegm heat, and heat toxin, demonstrating remarkable effects. The mechanism of blood sugar-lowering agents has always been a hotspot of research. Recently, the basic studies of heat-clearing prescriptions from different perspectives have been increasing year by year. To clarify the mechanisms of heat-clearing prescriptions and find specific mechanisms, we systematically reviewed the basic studies of heat-clearing prescriptions commonly used for the treatment of T2DM in the past decade, intending to provide a reference for related research.

Equivalence of combined decoction and mixed single decoctions of Gegen Qinlian Decoction in alleviating chemotherapy-associated diarrhea / 中国中药杂志

This study compared the chemical profiles, component content, dry paste yield, and pharmacological effects of samples obtained from the mixed single decoctions and the combined decoction of Gegen Qinlian Decoction(GQD), aiming to provide an experimental foundation for evaluating the equivalence of the two decocting methods and the suitability of TCM formula granules in clinical application. The same decoction process was used to prepare the combined decoction and mixed single decoctions of GQD. Ultra-performance liquid chromatography coupled with Q-Exactive Orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap MS) was employed to compare the chemical profiles between the two groups. High-performance liquid chromatography(HPLC) was used to compare the content of nine characteristic components between the two groups. Then, a delayed diarrhea mouse model induced by irinotecan was established to compare the pharmacological effects of the two groups on chemotherapy-induced diarrhea. The UPLC-Q-Exactive Orbitrap MS in ESI~+ and ESI~- modes identified 59 chemical components in the compound decoction and mixed single decoctions, which showed no obvious differences in component species. The content of baicalin and wogonoside was higher in the compound decoction, while that of puerarin, daidzein-8-C-apiosylglucoside, berberine, epiberberine, wogonin, glycyrrhizic acid, and daidzein was higher in the mixed single decoctions. Further statistical analysis revealed no significant difference in the content of the nine characteristic components between the compound decoction and the mixed single decoctions. The dry paste yield had no significant difference between the two groups. Compared with the model group, both compound decoction and mixed single decoctions alleviated the weight loss and reduced diarrhea index in mice. Both of them lowered the levels of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), cyclooxygenase-2(COX-2), intercellular adhesion molecule-1(ICAM-1), interleukin-10(IL-10), malondialdehyde(MDA), and nitric oxide(NO) in the colon tissue. Furthermore, they significantly increased the levels of glutathione peroxidase(GSH-Px) and superoxide dismutase(SOD). Hematoxylin-eosin(HE) staining showed that colon tissue cells were tightly arranged with clear nuclei in both groups without obvious difference. The compound decoction and mixed single decoctions showed no significant differences in chemical component species, content of nine characteristic components, dry paste yield, or the pharmacological effects on alleviating chemotherapy-induced diarrhea. The findings provide a reference for evaluating the flexibility and superiority of combined or single decocting method in the preparation of TCM decoctions or formula granules.

Study on serum pharmacodynamic material basis of Gegen Qinlian Decoction in treatment of ulcerative colitis based on UHPLC-Q-Orbitrap-MS / 中国中药杂志

This study established a mouse model of ulcerative colitis and explored the serum transitional components of Gegen Qinlian Decoction by UHPLC-Q-Orbitrap-MS. Based on the exact relative molecular weight and MS/MS spectrum, 55 prototype components and 59 metabolites were identified from the model group, while 18 prototype components and 35 metabolites from the control group. The prototype components in serum were mainly flavonoids and the characteristic components of the model group were alkaloids. Glucuronidation, sulfonation, and glycosylation have been confirmed to be the main metabolic types in vivo. The results of comparative analysis of differences indicated that puerarin, baicalin, wogonoside, wogonin, chrysin, oroxylin A, berberine, berberrubine, and palmatine were the characteristic components in model state, which at the same time, were confirmed by pharmacological studies to be the serum pharmacodynamic material basis of Gegen Qinlian Decoction in the treatment of ulcerative colitis. This study has provided reference for explaining the metabolic transformation pattern and mechanism of action of Gegen Qinlian Decoction in vivo.

Effects and mechanisms of Supplemented Gegen Qinlian Decoction Formula against podocyte pyroptosis and insulin resistance in model rats with diabetic kidney disease / 中国中药杂志

This study explored the in vivo effects and mechanisms of the modern classical prescription Supplemented Gegen Qinlian Decoction Formula(SGDF) against diabetic kidney disease(DKD). Sixty rats were randomly divided into the normal group, model group, SGDF group, and rosiglitazone(ROS) group. The modified DKD rat model was established by employing the following three methods: exposure to high-fat diet, unilateral nephrectomy, and intraperitoneal injection of streptozotocin(STZ). After modeling, rats in the four groups were treated with double distilled water, SGDF suspension, and ROS suspension, respectively, by gavage every day. At the end of the 6 th week of drug administration, all the rats were sacrificed for collecting urine, blood, and kidney tissue, followed by the examination of rat general conditions, urine and blood biochemical indicators, glomerulosclerosis-related indicators, podocyte pyroptosis markers, insulin resistance(IR)-related indicators, and key molecules in the insulin receptor substrate(IRS) 1/phosphatidylinositol-3-kinase(PI3 K)/serine threonine kinase(Akt) signaling pathway. The results showed that SGDF and ROS improved the general conditions, some renal function indicators and glomerulosclerosis of DKD model rats without affecting the blood glucose(BG). Besides, they ameliorated the expression characteristics and levels of podocyte pyroptosis markers, alleviated IR, and up-regulated the protein expression levels of the key molecules in IRS1/PI3 K/Akt pathway to varying degrees. In conclusion, similar to ROS, SGDF relieves DKD by targeting multiple targets in vivo. Specifically, it exerts the therapeutic effects by alleviating podocyte pyroptosis and IR. This study has preliminarily provided the pharmacological evidence for the research and development of new drugs for the treatment of DKD based on SGDF.

Molecular mechanism of Gegen Qinlian Decoction in promoting differentiation of brown adipose tissue to improve glucose and lipid metabolism disorders in diabetic rats / 中国中药杂志

This study explored the molecular mechanism underlying the Gegen Qinlian Decoction(GQD) promoting the differentiation of brown adipose tissue(BAT) to improve glucose and lipid metabolism disorders in diabetic rats. After the hypoglycemic effect of GQD on diabetic rats induced by high-fat diet combined with a low dose of streptozotocin was confirmed, the total RNA of rat BAT around scapula was extracted. Nuclear transcription genes Prdm16, Pparγc1α, Pparα, Pparγ and Sirt1, BAT marker genes Ucp1, Cidea and Dio2, energy expenditure gene Ampkα2 as well as BAT secretion factors Adpn, Fndc5, Angptl8, IL-6 and Rbp4 were detected by qPCR, then were analyzed by IPA software. Afterward, the total protein from rat BAT was extracted, and PRDM16, PGC1α, PPARγ, PPARα, SIRT1, ChREBP, AMPKα, UCP1, ADPN, NRG4, GLUT1 and GLUT4 were detected by Western blot. The mRNA expression levels of Pparγc1α, Pparα, Pparγ, Ucp1, Cidea, Ampkα2, Dio2, Fndc5, Rbp4 and Angptl8 were significantly increased(P<0.05) and those of Adpn and IL-6 were significantly decreased(P<0.05) in the GQD group compared with the diabetic group. In addition, Sirt1 showed a downward trend(P=0.104), whereas Prdm16 tended to be up-regulated(P=0.182) in the GQD group. IPA canonical pathway analysis and diseases-and-functions analysis suggested that GQD activated PPARα/RXRα and SIRT1 signaling pathways to promote the differentiation of BAT and reduce the excessive lipid accumulation. Moreover, the protein expression levels of PRDM16, PGC1α, PPARα, PPARγ, SIRT1, ChREBP, AMPKα, UCP1, GLUT1, GLUT4 and NRG4 were significantly decreased in the diabetic group(P<0.01), which were elevated after GQD intervention(P<0.05). Unexpectedly, the expression of ADPN protein in the diabetic group was up-regulated(P<0.01) as compared with the control group, which was down-regulated after the administration with GQD(P<0.01). This study indicated that GQD promoted BAT differentiation and maturity to increase energy consumption, which reduced the glucose and lipid metabolism disorders and thereby improved diabetes symptoms.

Preliminary study on effect of Gegen Qinlian Decoction on enzyme activity, gene expression and methylation level of FASN in adipose tissue of rats with insulin resistance / 中国中药杂志

To investigate the effect of Gegen Qinlian Decoction(GQD) on enzyme activity, gene expression and methylation level of fatty acid synthase(FASN) in adipose tissue from rats with insulin resistance induced by high-fat diet. The 60% fat-powered high-fat diet was continuously given to male SD rats to induce the insulin resistance model. Then, they were divided into five groups randomly and administrated by gavage every day for 16 weeks with following drugs respectively: 10 mL·kg~(-1)water for control group(C) and insulin resistance model control group(IR), 1.65 g·kg~(-1)GQD per day for low-dose group(GQDL), 4.95 g·kg~(-1)GQD per day for medium-dose group(GQDM), 14.85 g·kg~(-1)GQD per day for high-dose group(GQDH), and 5 mg·kg~(-1) rosiglitazone per day for rosiglitazone group(RGN). Epididymal adipose tissue was taken to determine enzyme activity of FASN by colorimetric method, mRNA expression level of Fasn by quantitative Real-time PCR(Q-PCR) and CpGs methylation level between +313 and +582 by bisulfite sequencing PCR(BSP). These results showed that Fasn expression was significantly lowered in IR model rats compared with the control rats(P<0.01). Enzymatic activity and CpGs methylation level of Fasn in IR group showed downward trends. Low and medium-dose GQD can increase enzyme activity of FASN(P<0.05). Moreover, low-dose GQD increased the total CpGs methylation level of Fasn fragment between +313 and +582 in insulin resistance rats(P<0.05). For GQDM group, the methylation frequency of CpGs at positions +506 and +508(P<0.01) as well as the methylation frequency of CpGs on the binding sites of transcription factorzinc finger protein 161(P<0.05) were significantly increased. The methylation frequency of CpG at +442 position was positively correlated with Fasn expression(P<0.01, r=0.735), and methylation frequencies of CpGs at +345 and +366 positions were positively associated to enzyme activity of FASN respectively(P<0.05, r=0.479; P<0.01, r=0.640). In conclusion, GQD can reverse enzyme activity of FASN and methylation level of Fasn in adipose tissue of insulin resistant rats, and CpG sites at positions +506 and +508 may be the targets of GQD. The methylation level of CpGs at + 345 and + 366 sites were possibly related to FASN activity, while methylation of CpG at + 442 site may be closely correlated with mRNA level of Fasn. In addition, GQD did not significantly change mRNA expression level of Fasn, but effectively reversed enzymatic activity, suggesting that GQD may regulate the post transcriptional expression of Fasn.

Chemical constituent analysis of Gegen Qinlian Decoction based on UPLC-LTQ-Orbitrap-MS / 中草药

Objective: Identification of chemical constituents from Gengen Qinlian Decoction by UPLC-LTQ-Orbitrap-MS. Methods: The analysis was performed on Dikma Endeavorsil C18 (100 mm × 2.1 mm, 1.8 μm) with mobile phase of 0.1% formic acid water solution (A)-acetonitrile (B) for gradient elution at a flow rate of 0.3 mL/min. The UPLC-LTQ-Orbitrap-MS was equipped with an Electrospray ionization ion probe and MS1 and MS2 data of samples were collected in positive and negative ion mode, respectively. Results: A total of 67 constituents were identified from Gegen Qinlian Decoction by reference substance identification, software prediction analysis and related literature reports, including 36 flavonoids, 12 alkaloids, four triterpenoids and triterpenoid saponins, and 15 other ingredients. Conclusion: In this study, UPLC-LTQ-Orbitrap-MS was used to systematically elucidate the chemical constituents of Gegen Qinlian Decoction, and the fragmentation characteristics of its main chemical constituents were preliminarily explained and summarized, which provided a reference for the quality control and mechanism research of Gegen Qinlian Decoction.

Effect of Gegen Qinlian Decoction and composition of index components on ulcerative colitis related colon cancer in mice based on "pathway-disease" interaction network / 中草药

Objective: A network model of ulcerative colitis-associated colon cancer (UCRCC) and NF-κB signaling pathway was established, in order to predict the key inflammatory targets of UCRCC and identify the effect of GQD and composition of index components on these targets. Methods: HPLC method was used, the mobile phase was acetonitrile-0.2% phosphoric acid aqueous solution, and the gradient elution was carried out. The 10 indicators were determined. The literature of databases such as Pubmed and ScienceDirect were searched, the terms of the upstream and downstream proteins of UCRCC disease and NF-κB pathway were examined. The Cytoscape software was used to predict the key inflammatory targets of UCRCC. The UCRCC model was established by AOM/DSS method, and the effect of GQD and composition of index components on key inflammatory targets were identified by immunohistochemistry and ELISA. Results: The contents of puerarin, daidzin, glycyrrhizin, jatrorrhizine, baicalin, palmatine, berberine, baicalein, glycyrrhizic acid and wogonin in GQD were 1.677, 0.154, 0.159, 0.045, 0.448, 0.035, 0.095, 0.013, 0.111 and 0.006 μg/g, respectively. The top eight inflammatory factors of the key protein in the interaction network were NF-κBp65, iNOS, COX-2, Bcl-2, TNF-α, IL-1β, ICAM-1 and VCAM-1. Compared with the model group, the high-, medium- and low-dose groups of GQD, the composition of index components group and the 5-ASA group were able to down-regulate the relative expression levels of NF-κBp65, iNOS, Bcl-2, COX-2 in colonic tissues of UCRCC mice (P < 0.01), and down-regulate the expression levels of TNF-α, IL-1β, ICAM-1, VCAM-1 in serum of UCRCC mice (P < 0.05, 0.01). Conclusion: GQD and composition of index components can improve the pathological condition of colon tissue in UCRCC mice, down-regulate the expression of inflammatory factors in colon tissue of UCRCC mice, and have a certain intervention effect on UCRCC, which laid the foundation for determining the quality marker (Q-marker) of GQD.

Gegen Qinlian Decoction formula syndrome and its application in diabetes, hypertension, hyperlipidemia and obesity / 中国中药杂志

Gegen Qinlian Decoction can be used to treat intestinal dampness and heat. In addition to diarrhea diseases, it is also commonly used in the treatment of diabetes, hypertension, hyperlipidemia, obesity and other chronic metabolic diseases. It can not only alleviate symptoms, but also reduce blood sugar, blood pressure, lipid and weight. Neck stiffness, blush, red lips, red tongue, dry mouth, sweating, palpitation, insomnia and feces are the key indications of Gegen Qinlian Decoction. It can be used alone to reduce blood sugar for diabetes mellitus. In the treatment of hypertension, it can reduce blood pressure when being used alone or combined with Tianma Gouteng Yin or Chaihu Jia Longgu Muli Decoction. Large dose(30-120 g) of Pueraria lobata is the key to the effect of Gegen Qinlian Decoction.

Study on the Mechanism of Gegen Qinlian Decoction for Lowering Blood Lipids and Preventing Blood Glucose Increase Based on Intestinal Flora / 中国药房

OBJECTIVE：To study the ef fects of Gegen qinlian decoction （GGQLD）on blood lipid and blood glucose of hyperlipidemia（HLP）model rats ，and to explore its mechanism from the perspective of intestinal flora. METHODS ：Totally 48 rats were randomly divided into blank control group （n＝8）and modeling group （n＝40）. For consecutive 5 weeks，model group was given high-lipid diet to induce HLP model ；blank control group was given routine diet. After modeling ，30 modeling rats were randomly divided into model group ，simvastatin group （positive control ，10 mg/kg），GGQLD high-dose ，medium-dose and low-dose groups （14.85，4.95，1.65 g/kg，by crude drug ），with 6 rats in each group. Blank control group and model group were given constant volume of normal saline intragastrically ；administration groups were given relevant medicine intragastrically ，once a day，for consecutive 11 weeks. At the same time ，each group was continuously given corresponding diet. After the last medication ， body mass and body length of rats were determined ，and Lee ’s index was calculated. Serum levels of TG ，TC，HDL-C，LDL-C and fasting blood glucose （FBG）were determined in rats. DNA of rat caecum content was extracted for 16S rRNA V 3-V4 region sequencing. The Two-part model was used to analyze the correlation between intestinal flora with lipids and blood glucose. RESULTS：After 11 weeks of administration ，compared with blank control group ，the body mass ，body length ，Lee’s index ， serum levels of TC ，TG，HDL-C and FBG of model group were increased significantly （P＜0.05 or P＜0.01），while the level of HDL-C was decreased significantly （P＜0.05）. Compared with model group ，body mass and Lee ’s index and serum levels of TG ， FBG of rats in GGQLD high-dose group ，and serum levels of TC ，TG in GGQLD medium-dose group ，as well as serum level of TG of rats in GGQLD low-dose group was decreased significantly （P＜0.05 or P＜0.01）. Correlation analysis with intestinal flora showed that TC and TG shared 3 operational taxonomic units （OTU），including OTU 559，OTU701 and OTU 135（OTU135 was also shared with FBG ），which were all positively correlated with the level of TC ，TG and FBG （P＜0.01）. The three OTU were annotated as Tyzzerella of Spirillaceae ，Anaerotruncus of Verrucaceae and Peptoclostridium of Streptococcidae ，respectively. High-dose and low-dose GGQLD had a down-regulating effect on Tyzzerella and Anaerotruncus（P＜0.05 or P＜0.01），while had up-regulating effect on Peptoclostridium（P＜0.01）. CONCLUSIONS ：High-dose GGQLD （14.85 g/kg）can effectively reduce the body mass and blood lipid of HLP model rats ，and can prevent the abnormal increase of blood glucose of model rats. The mechanism may be associated with that the reduction of intestinal flora （Tyzzerella，Anaerotruncus）content.

Study on raw and fermented Gegen Qinlian Decoction for treatment of diabetes based on 1H-NMR metabolomics in T2DM rats / 中草药

Objective To compare the antidiabetic effects between raw and fermented Gegen Qinlian Decoction (GQD) containing different endogenous metabolites by 1H NMR technology based on metabolomics. Methods The antidiabetic effects were evaluated using high glucose and high fat diet combined with streptozotocin (STZ). The treatment groups were fed with GQD and fermented GQD every day, respectively for 8 weeks.The general condition, body weight and fasting blood glucose (FBG) of rats were observed. The chemical compositions of serum were assigned by 1H NMR spectroscopy. Using the multivariate statistical analysis as the main data analysis methods, potential biomarkers were screened in the model. Results The body weight of the diabetic rats decreased significantly compared to the normal control group after injection of STZ. In addition, the FBG level was significantly increased in the diabetic rats than that in the normal control group. GQD and fermented GQD all can improve the general condition, body mass, and FBG level of T2DM rats. The results of the principal component analysis showed that the metabolic profile of the normal control group, the model group and drug treatment group was significantly differentiated. In total, 15 potential biomarkers associated with T2DM were identified by OPLS-DA binding univariate analysis. The corresponding S-plot combined with VIP > 1 revealed that the major variations in diabetic rats were the elevation of 3-hydroxybutyrate (3-HB), choline, glycine, glycerol, β-glucose, and α-glucose, as well as the decline levels of lactate, very low-density lipoprotein (VLDL), trimethylamine-N-oxide (TMAO), acetate, glutamate, methionine, glutamine, pyruvate, and creatine. Both raw and fermented GQD displayed antidiabetic effects against STZ-induced diabetes by restoring 15 of biomarkers. Conclusion Detailed analysis of the altered metabolite levels indicated that raw and fermented GQD significantly ameliorated the disturbance in glucose metabolism, lipid metabolism, and amino acid metabolism in the therapeutic process of T2DM, while no significant difference was observed between them. The results of this study provide experimental data and theoretical basis for the clinical application of GQD fermentation from the perspective of metabolomics.

Experience in Treating Eosinophilic Gastroenteritis with Banxia Xiexin Decoction / 中国中医药信息杂志

The most common manifestations of eosinophilic gastroenteritis (EG) are abdominal pain and diarrhea. Glucocorticoid is currently used in the treatment for EG, but the adverse reactions are relatively large. Banxia Xiexin Decoction is the famous Zhongjing prescription can be used to treat many kinds of gastrointestinal diseases. This article introduced. Banxia Xiexin Decoction combined with Shaoyao Gancao Decoction in the treatment of abdominal pain predominant based EG, as well as Banxia Xiexin Decoction combined with Gegen Qinlian Decoction in treating diarrhea predominant based EG have achieved satisfactory efficacy.

Effect of Puerarin on the Pharmacokinetics of Baicalin in Gegen Qinlian Decoction () in Mice / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the pharmacokinetics of puerarin (PUE) in Gegen Qinlian Decoction (, GQD), and the effects of PUE dosage variations on the pharmacokinetics of baicalin (BAL) in mice.</p><p><b>METHODS</b>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 (icELISA) 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.</p><p><b>RESULTS</b>After the co-administration of GQD with PUE, the area under the curves (AUC) of PUE increased 2.8 times compared with GQD. At the dose of GQD co-administration at two times that of PUE, the (AUC) of PUE was almost equal to that of GQD co-administration of PUE, showing non-linear pharmacokinetics. The (AUC) 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 (T), the second T, or the mean residence time.</p><p><b>CONCLUSIONS</b>The icELISA 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.</p>

Effect of different concentrations of Gegen Qinlian decoction on expression of P-glycoprotein and multi-drug resistance protein transporter in Caco-2 cells / 中国中药杂志

To investigate the effects of Gegen Qinlian decoction on the expression of P-glycoprotein (P-gp) and multi-drug resistance protein (MRP) in epithelial cells of human colon adenocarcinoma Caco-2 cells．The effects of different concentrations of Gegen Qinlian decoction on the expression levels of p-gp and MRP1-6 mRNA in Caco-2 cells were detected by real time quantitative reverse transcriptase PCR (qRT-PCR).12 h after drug treatment (5.00 g·L⁻¹), the expression levels of MDR1 and MRP1-6 were significantly down-regulated at concentration of 5.00 g·L⁻¹; the mRNA expression levels of MDR1，MRP1，MRP2，MRP4，MRP5 and MRP6 were significantly down-regulated at concentration of 2.50 g·L⁻¹; only the expression levels of MRP2 and MRP5 were significantly affected at concentration of 1.00 g·L⁻¹. The results showed that the expression levels of MDR1 and MRP1-6 mRNA in Caco-2 cells could be down-regulated in a dose-dependent manner. Gegen Qinlian decoction may reduce drug efflux by down-regulating the mRNA expression of cell transporters in Caco-2 cell, and increase the time of drug action, thereby enhancing the bioavailability of chemotherapeutic drugs.

Pharmacokinetic characteristics of fourteen main components of Gegen Qinlian Decoction in rats / 中国中药杂志

A specific and selective UPLC-MS/MS method was developed and validated for the simultaneous determination of isoflavonoids(3'-hydroxy puerarin, puerarin, daidzin, daidzein, genistin, genistein), flavonoids (baicalin, baicalein, wogonoside, wogonin, liquiritin)and alkaloids(berberine, jatrorrhizine, palmatine)(14 bioactive compounds) of Gegen Qinlian Decoction(GQD) in plasma. The pharmacokinetics characteristics of 14 bioactive compounds were study after oral administration of GQD at a single dose to rats. Prednisolone was used as the internal standard of liquiritin, and naringin was used as the internal standard of the other thirteen analytes. After the plasma samples were processed by precipitation protein method, the constituents and internal standards were gradient eluted by using a Zorbax SB-18 column with a mobile phase of acetonitrile(A) and 0.1% formic acid aqueous solution(B) using a gradient elution of 0-2.5 min, 15%-30% A; 2.5-3.5 min, 30%-35% A; 3.5-5.0 min, 35%-40% A; 5.0-9.0 min, 40%-60% A; 9.0-11.0 min, 60%-15% A, and the flow rate was 0.4 mL·min⁻¹. The auto sampler was conditioned at 25 °C and the sample injection volume was 5 μL. A mass spectrometry was applied with electrospray ionization (ESI) ion source in the positive and negative ion multiple reaction monitoring(MRM) mode. All pharmacokinetic parameters were processed by non-compartmental analysis with DAS 3.2.2 software. The results showed that the linear correlation coefficient of the 14 components were all greater than 0.99, indicating that the method had good linearity in their respective concentration ranges. Post-preparative stability (25 °C, 24 h), short-term stability(25 °C, 12 h), long-term stability (-20 °C, 7 d), and freeze and thaw stability (3-cycles) of the fourteen constituents were examined to evaluate the stability of methodology. The results of the inner and inter-day relative standard deviations were both less than 10%, indicating legitimate precise and accuracy to the requirement of biological sample analysis. The assay method is proved to be sensitive, accurate and convenient. It can be applied to the pharmacokinetic study of the fourteen analytes. The kinetic parameters of the related drugs were calculated according to the blood concentration of the 14 components. The results showed that the MRT0-t of the isoflavones and flavonoids was 7.5-11.8 h, T1/2z were mainly in 11.0-29.7 h, and the AUC0-t flavonoids were larger than the isoflavones. The MRT0-t of alkaloids were between 4.3-7.2 h, T1/2z were 1.0-5.0 h, AUC0-t were less than flavonoids and isoflavones. The results suggest that flavonoids and isoflavones have a high concentration of blood and long time of action, which are beneficial to the anti-inflammatory and antipyretic effects. The concentration of alkaloids in the body is low and the time of action is short, and it may play its bacteriostasis in the intestinal tract.

Comparative Study of Probiotics and Traditional Chinese Medicine Syndrome-differentiation Treatment for Rotavirus Enteritis in Children / 广州中医药大学学报

Objective To investigate the therapeutic effect of probiotics and traditional Chinese medicine syndrome-differentiation treatment for children rotavirus enteritis. Methods One hundred and twenty cases of children with rotavirus enteritis were randomly divided into Chinese medicine group, probiotics group and control group,40 cases in each group. The 3 groups were given conventional treatment such as proper feeding, prevention of dehydration and correction of dehydration. Additionally, Chinese Medicine group was separately given modified Gegen Qinlian Decoction for patients with damp-heat syndrome, modified Huoxiang Zhengqi Decoction for patients with wind-cold syndrome, modified Baohe Pills for patients impaired by overeating, modified Shenling Baizhu Powder for patients with spleen deficiency syndrome;probiotics group received Probiotic Bifidobacterium Triple Viable Enteric Capsules;the control group was given oral use of Smecta. Daily defecation frequency and stool characteristics were recorded,and the clinical efficacy was analyzed on treatment day 3 and 5. Results(1)After treatment for 3 days,the total effective rate of the 3 groups arrived to 100%,the difference being insignificant between the 3 groups (P > 0.05);after treatment for 3 days, Chinese medicine group had higher markedly effective rate than probiotics group and control group,the difference being significant (P < 0.05). (2) After treatment for 3 and 5 days,the defecation frequency of the 3 groups was much reduced as compared with that before treatment (P < 0.05), and the improvement of defecation frequency in Chinese medicine group was superior to that in probiotic group and control group, the difference being significant (P <0.05). (3)The mean course of disease in Chinese medicine group was shorter than that in probiotic group and control group,the difference being significant (P < 0.05). Conclusion Traditional Chinese medicine syndrome-differentiation treatment exerts certain effect for the treatment of rotavirus enteritis, and can quickly reduce defection frequency and shorten the course of disease.

Effect of Gegen Qinlian Decoction on LPS, TNF-α, IL-6, and intestinal flora in diabetic KK-Ay mice / 中草药

Objective: To explore the effect of Gegen Qinlian Decoction (GGQLD) on LPS, TNF-α, IL-6, and intestinal flora in diabetic KK-Ay mice. Methods: C57BL/6J mice with ordinary feed were taken as the normal control group and orally administrated with equal distilled water. The KK-Ay mice fed with high-fat diet were divided into five groups: pioglitazone group, blank group (model group), high, medium, and low dose GGQLD group, and orally administrated with pioglitazone hydrochloride (5 mg/kg), distilled water, and GGQLD (crude drug 40, 13.3, and 4.44 g/kg), respectively. The oral administration for six groups lasted for four weeks. Tumor necrosis factor (TNF-α), interleukin 6 (IL-6), and endotoxin (LPS) levels in the plasma were determined by enzyme-linked immunosorbent assay (ELISA); Gut microbial communities were assayed by polymerase chain reaction (PCR) and PCR-denaturing gradient gel electrophoresis (PCR-DGGE) methods. Results: Compared with the model group, the LPS levels in the plasma of mice were significantly reduced by 15.61% and 14.48% respectively in the Gegenqinlian Decoction of high and medium dose group (P < 0.05), the IL-6 levels in plasma of mice were significantly reduced by 56.86%, 37.12% and 30.21% respectively in high, medium, and low dose GGQLD group (P < 0.05), and the TNF-α levels in plasma of mice were significantly reduced by 28.32%, 30.70%, and 23.42% respectively in high, medium, and low dose GGQLD group (P < 0.05). The number of DGGE bands in high dose group significantly increased, and by cloning, sequencing, and Blast analysis, Lactobacillus johnsonii only existed in the high dose group; The results showed that GGQLD could regulate the structure of intestinal flora in KK-Ay mice. Conclusion: The mechanisms of anti-diabetic effects of GGQLD in type 2 diabetic KK-Ay mice are probably related with the anti-inflammation and regulation of intestinal flora.

Comparative study of Chinese medicine Gegen Qinlian Decoction Piece and boiled powder for the treatment of diabetes based on serum metabolomics / 药学学报

To investigate the difference of Gegen Qinlian Decoction (GQD) piece and boiled powder in the treatment of type 2 diabetes (T2DM), the characteristic of overall metabolite profile was examined in the serum of T2DM rats with 1H NMR-based metabolomics combined with the multivariate statistical analysis. A rat model of T2DM was established by feeding of high glucose and high fat diet followed by a streptozotocin (STZ) treatment. The general condition, body weight and fasting blood glucose (FBG) of rats were monitored. GQD piece and boiled powder exhibited activities in the improvement of these parameters. The results of the principal component analysis showed that there was a significant difference in the metabolic profile of the normal control group, the model group, the positive group, the herbal decoction group and the boiled powder group. Totally 15 potential biomarkers were identified by OPLS-DA binding univariate analysis. Compared with normal control group, the serum samples of T2DM showed a higher level of 3-HB, TMAO, glycine, β-glucose and α-glucose accompanied by lower level of lactate, VLDL, acetate, glutamate, methionine, glutamine, pyruvate, creatine, choline and glycerol. The above results also demonstrated that both piece and boiled powder of GQD could restore 14 of these markers. These results suggested that the disrupted metabolic pathways including energy metabolism, lipid metabolism and amino acid metabolism, were restored by GQD piece and boiled powder. The two formula did not show a significant difference. The results of this study provide experimental data and theoretical basis for the equal activities of GQD piece and boiled powder in clinical application.

Gegen Qinlian decoction activates PPARγ to ameliorate adipocytic insulin resistance in diabetic SD rats and IR-3T3-L1 adipocytes / 中国中药杂志

To investigate the effects of Gegen Qinlian decoction(GQD) in improving adipocytic insulin resistance(IR) and explore its related molecular mechanism. Diabetic rats models were induced by high glucose and high-fat diet with a small dose of streptozotocin, and after GQD treatment for 3 months, blood biochemical indexes such as fasting blood-glucose(FBG), insulin, glycosylated serum protein(GSP) and HOMA-IRI were detected and assessed. After the total RNA was extracted from the adipose tissue of diabetic SD rats, PPARγ, ADPN, GLUT4, GLUT2, ACACA and ACACB mRNA expression levels were separately detected by qPCR. Then, stable IR-3T3-L1 adipocyte model was built with 1 μmol•L⁻¹ dexamethasone. After the cell viability was detected by CCK-8 assay, 5%, 10% and 15% GQD-containing serum(GQD-CS) were respectively used to treat IR-3T-L1 adipocytes for 24 h. The contents of glucose, nonesterified fatty acid(NEFA) and adiponectin in cell culture supernatants were separately detected whereas the intracellular triglyceride(TG) contents of IR-3T3-L1 adipocytes were also measured. The ADPN, PPARγ and GLUT4 mRNA and protein expression levels were respectively detected by qPCR and Western blot in IR-3T3-L1 adipocytes. Results showed that GQD significantly decreased fasting blood glucose, insulin and GSP(P<0.01), and down-regulated HOMA-IRI(P<0.05) after the high-fat diet/streptozotocin-induced diabetic SD rats were treated for three months, with a good hypoglycemic effect. Moreover, PPARγ, ADPN, GLUT4, GLUT2, ACACA and ACACB mRNA expression levels were significantly elevated in the adipose tissue of GQD-treated diabetic SD rats. The 5%, 10% and 15% GQD-CS significantly increased glucose consumption of IR-3T3-L1 adipocytes at 24 h treatment(P<0.01), significantly decreased the intracellular TG content (P<0.01), and down-regulated NEFA to a certain extent but not significantly. Moreover, GQD-CS significantly up-regulated GLUT4 and ADPN expression. The results indicated that GQD could activate PPARγ to ameliorate adipocytic insulin resistance in the diabetic SD rats and IR-3T3-L1 adipocytes.