Action mechanism of Sijunzi Decoction in treatment of type 2 diabetes mellitus based on network pharmacology / 中草药

Objective: The main active components and targets of Sijunzi Decoction for type 2 diabetes mellitus (T2DM) treatment were predicted by network pharmacology, and the potential mechanism of multi-component-multi-target-multi-pathway was discussed, and INSR and PI3K in the AMPK signal pathway were verified in animal experiment. Methods: Based on TCM system pharmacology database and analysis platform (TCMSP), literature mining and previous studies of our research group, the chemical components of Sijunzi decoction and its four herbs Codonopsis pilosula, Atractylodes macrocephala, Poria cocos, Glycyrrhiza uralensis were screened. Three conditions of oral absorption rate (OB), drug-like (DL), half-life (HL) were used to screen the active components of the drug; The targets of active components were predicted through the Swiss Target Prediction database; TTD, Drupe and DisGNET databases were used to screen the targets of T2DM. After mapping the component target and disease target, the interaction network of the target proteins of the active components was constructed by using the software of Cytoscape 3.2.1. Through topological analysis of the network, the nodes with degree of freedom ≥ average degree of freedom were screened out, and the core target network of core active ingredient was obtained, while using the String database to draw core-target protein-protein interaction (PPI) networks, using DAVID database for GO analysis and KEGG analysis of core target genes to construct core active components-core target-metabolism Path network diagram to explore the potential mechanism of Sijunzi Decoction against T2DM. The system docking site was used for molecular docking of the top 5 components and the top 3 proteins. Further, animal experiments were used to verify. SD rats were fed with high-sugar high-fat diet combined with low-dose streptozotocin (STZ) to induce T2DM model, and administrated with 5.65 g/kg Sijunzi Decoction for 28 d, and the levels of blood glucose (FBG) and oral glucose tolerance (OGTT) of each group of mice were determinated. The InsR, PI3K mRNA expression in AMPK signaling pathway was detected by RT-PCR. Results: A total of 113 chemical components were selected from Sijunzi Decoction, involving 47 targets for the treatment of T2DM. Twenty-two core components and 27 core targets were screened according to node degree of freedom ≥ average degree of freedom. The GO analysis results showed that it involved seven biological processes such as blood glucose homeostasis, positive regulation of adipose tissue development, four molecular functions including steroid hormone receptor activation and drug binding, and cell composition including the plasma membrane and nuclear chromatin. The results of KEGG analysis showed that it might treat diabetes through 14 signaling pathways, such as AMPK signaling pathway, PPAR signaling pathway and insulin resistance. A total of 60% of the molecules had intense binding activity and 40% had stronger binding activity. The results of animal experiments showed that Sijunzi Decoction could significantly improve OGTT (P < 0.001), and decrease FBG (P < 0.01) in T2DM rats, and significantly increase the expression of INSR mRNA (P < 0.001) and PI3K mRNA (P < 0.001). Some of the predicted results of network pharmacology were verified. Conclusion: This study reflects that Sijunzi Decoction can treat type 2 diabetes mellitus through multi-target and multi-channel, which lays a foundation for the future study of molecular mechanism.

Effects of Zuogui Jiangtang Jieyu Formulation on insulin resistance in hipppocampus of rats with diabetes-related depression / 中草药

Objective: To investigate the function of Zuogui Jiangtang Jieyu Formulation on hippocampal insulin resistance in rats with diabetes-related depression. Methods: The rat model of diabetes-related depression was established and randomly divided into four groups, including model group, positive drug group (metformin 0.18 g/kg + fluoxetine 1.8 mg/kg), high (20.53 g/kg) and low (10.26 g/kg) doses of Zuogui Jiangtang Jieyu Formulation groups. After 28 d of gavage, fasting blood glucose and peripheral insulin resistance were measured by blood glucose meter and ELISA. The depression-like behaviors of rats were tested by open field experiment and forced swimming test. The expression of p-IR and p-IRS-1 in hippocampus of rats were detected by immunofluorescence and Western blotting, while the levels of p-PI3K and p-Akt were detected by Western blotting. Results: Compared with the normal group, the blood glucose of rats in model group were significantly increased, which accompanied by obvious peripheral insulin resistance. The number of activities in open field experiment was significantly reduced in model group, and the immobility time in forced swimming experiment was significantly prolonged. In addition, it was showed that the levels of p-IR, p-IRS-1, p-PI3K and p-Akt in the brain of model rats were all significantly decreased. Compared with the model group, the blood glucose and insulin levels were significantly decreased in high-dose of Zuogui Jiangtang Jieyu Formulation group. Furthermore, the depression-like behaviors manifested in open field experiment and forced swimming experiment were improves by high dose of Zuogui Jiangtang Jieyu Formulation. More importantly, the expression of p-IR, p-IRS-1, p-PI3K and p-Akt in hippocampus was significantly increased in high dose of Zuogui Jiangtang Jieyu Formulation group compared with model group. Conclusion: Zuogui Jiangtang Jieyu Formulation can effectively regulate the insulin signaling pathway in hippocampus of rats with diabetes-related depression, and then improve the insulin resistance in the brain of model rats.

Palmitic acid-induced lipotoxicity promotes a novel interplay between Akt-mTOR, IRS-1, and FFAR1 signaling in pancreatic ß-cells

BACKGROUND: Free fatty acid receptor 1 (FFAR1) is G-protein coupled receptor predominantly expressed in pancreatic ß-cells that is activated by a variety of free fatty acids (FFAs). Once activated, it promotes glucose-stimulated insulin secretion (GSIS). However, increased levels of FFAs lead to lipotoxicity, inducing loss of ß-cell function. FFAR1 plays a key role in the development of type 2 diabetes (T2D), and previous studies have indicated the importance of developing anti-diabetic therapies against FFAR1, although its role in the regulation of ß-cell function remains unclear. The present study investigated the role of FFAR1 under lipotoxic conditions using palmitic acid (PA). The rat insulinoma 1 clone 832/13 (INS-1 832/13) cell line was used as a model as it physiologically resembles native pancreatic ß-cells. Key players of the insulin signaling pathway, such as mTOR, Akt, IRS-1, and the insulin receptor (INSR1ß), were selected as candidates to be analyzed under lipotoxic conditions. RESULTS: We revealed that PA-induced lipotoxicity affected GSIS in INS-1 cells and negatively modulated the activity of both IRS-1 and Akt. Reduced phosphorylation of both IRS-1 S636/639 and Akt S473 was observed, in addition to decreased expression of both INSR1ß and FFAR1. Moreover, transient knockdown of FFAR1 led to a reduction in IRS-1 mRNA expression and an increase in INSR1ß; mRNA. Finally, PA affected localization of FFAR1 from the cytoplasm to the perinucleus. CONCLUSIONS: In conclusion, our study suggests a novel regulatory involvement of FFAR1 in crosstalk with mTOR-Akt and IRS-1 signaling in ß-cells under lipotoxic conditions.

Effects of Compound Malt Pill on Gene Expressions of Androgen Receptor and Insulin Receptor in Polycystic Ovarian Syndrome Rat Models / 中国中医药信息杂志

Objective To explore the effects of Compound Malt Pill on gene expressions of androgen receptor (AR) and insulin receptor (INSR) in polycystic ovarian syndrome (PCOS) rats; To discuss its relevant mechanism of action.Methods Letrozole was used to induce and establish rat models. Rats were randomly divided into normal group, model group, Diane-35 group, low-, medium- and high-dose Compound Malt Pill groups. The normal group and model group received normal saline by gavage; the Diane-35 group was given Diane-35 by gavage; Compound Malt Pill groups were respectively given different concentrations by gavage for 21 d. The levels of serum testosterone (T) and insulin were measured by ELISA, and the gene expressions of AR and INSR were measured by RT-PCR. Results Compared with normal group, the serum T, insulin and the gene expressions of AR and INSR in model group increased significantly (P<0.05,P<0.01); compared with model group, the serum T, insulin and the gene expressions of AR and INSR in low-, medium- and high-dose Compound Malt Pill groups decreased significantly (P<0.05, P<0.01).Conclusion Compound Malt Pill can adjust endocrine-metabolic disorder in PCOS rats.

Two Novel Insulin Receptor Gene Mutations in a Patient with Rabson-Mendenhall Syndrome: The First Korean Case Confirmed by Biochemical, and Molecular Evidence

Rabson-Mendenhall syndrome (RMS) is a rare syndrome manifested by extreme insulin resistance with hyperinsulinemia, acanthosis nigricans, tooth dysplasia and growth retardation. Our patient was first noted at the age of 8 months due to pigmentations on skin-folded areas. Initial laboratory tests showed normal fasting glucose (69 mg/dL). Fasting insulin level was severely elevated, up to 554.6 microIU/mL, and c-peptide level was increased, up to 13.81 ng/mL. However, hemoglobin A1c was within normal range (4.8%). He is now 11 yr old. His growth development followed the 5-10th percentile and oral hypoglycemic agents are being administered. The last laboratory results showed insulin 364.1 microIU/mL, C-peptide 4.30 ng/mL, and hemoglobin A1c 7.6%. The boy was a compound heterozygote for the c.90C > A and c.712G > A mutations of the insulin receptor gene, INSR, which are nonsense and missense mutations. In summary, we report the first Korean case of RMS, which was confirmed by two novel mutations of the INSR.

Effect of IGF-1 on the InsR?、? gene expression and protein content in pyaemia rats / 肠外与肠内营养

Objective:To investigate the influences of IGF-1 on the stress homone,InsR?、? gene expression and protein content in pyaemia rats.Methods:SD rats with abdominal infection models established by cecal ligation and perforation method were randomly divided into three groups: pyaemic group(n=10),parenteral nutrition group(PN group,n=10) and IGF-1 Group (n=10),and 10 SD rats were used as normal group(n=10).On the sixth day,Vena Cava blood was sampled to determine the level of glucose,insulin,glucagon,and IGF-1.In addition,the expression of the InsR?、? in liver and muscle were detected by immunohistochemistry and Real-Time PCR methods.Results:The plasma glucose,insulin,glucagon in the IGF-1 group were significantly lower than that of the pyaemic group and PN group(P