Mechanism of Huanglian Wendan Decoction in improving impaired glucose tolerance based on skeletal muscle NLRP3/caspase-1/IL-1β, IL-18 pathway / 中国中药杂志

This study investigated the mechanism of improving impaired glucose tolerance(IGT) of rats by Huanglian Wendan Decoction from the perspective of the skeletal muscle Nod-like receptor protein 3(NLRP3)/cysteinyl aspartate specific proteinase-1(caspase-1)/interleukin-1β(IL-1β), interleukin-18(IL-18) pathway. Healthy male SD rats were fed with the diet containing 45% fat for 20 weeks, accompanied by a high-temperature and high-humidity environment and an inactive lifestyle, for the establishment of the IGT rat model. The rats were divided into the blank control group, model control group, metformin hydrochloride group(positive drug group, 0.05 g·kg~(-1)·d~(-1)) and Huanglian Wendan Decoction group(7.8 g·kg~(-1)·d~(-1)). After continuous intragastric administration for 4 weeks, the obesity and glycemic indexes of all the rats were measured. The fasting serum insulin(FINS) level was determined by ELISA, with the insulin sensitivity index(ISI) and insulin resistance index(IRI) calculated. The mRNA and protein expression le-vels of nuclear factor kappaB(NF-κB), NLRP3, caspase-1, IL-1β and IL-18 in skeletal muscle tissue were detected by real-time polymerase chain reaction(PCR), Western blot and immunofluorescence. Compared with the blank control group, the model control group witnessed significantly increased mRNA and protein expression of NF-κB, NLRP3, caspase-1, IL-1β and IL-18. As revealed by the comparison with the model control group, Huanglian Wendan Decoction could effectively regulate the obesity status, reduce body weight, correct the abnormal levels of 2-hour plasma glucose(2 hPG), insulin resistance index(IRI), insulin sensitivity index(ISI), and lower the mRNA and protein expression of NF-κB, NLRP3, caspase-1, IL-1β and IL-18 in the skeletal muscle tissue of IGT rats. Combined with previous studies, the above results showed that the occurrence and development of IGT was closely related to inflammatory response and the classic pyroptosis pathway in skeletal muscle, such as NLRP3/caspase-1/IL-1β, IL-18. It is inferred that the mechanism of Huanglian Wendan Decoction was to alleviate insulin resistance(IR) and then reverse the course of IGT lies in the regulation of the abnormal insulin receptor signaling pathway based on the NLRP3 inflammasome pathway.

Effect of Huanglian Wendantang on Inflammatory Response and Hepatocyte Burn Death in IGT Rats / 中国实验方剂学杂志

Objective:To investigate whether there is inflammatory reaction and cell pyroptosis in impaired glucose tolerance （IGT） rats induced by high-fat diet and the intervention effect of Huanglian Wendantang. Method:Healthy male SD rats were fed with 45% fat content diet for 20 weeks to replicate the IGT model. The rats in line with the model establish criteria were randomly divided into 3 groups， with 10 rats in each group， another 10 rats were selected as the blank control group. Huanglian Wendantang group was given 7.8 g·kg^-1·d^-1 compound decoction of Huanglian Wendantang， and the positive control group was given 0.05 g·kg^-1·d^-1 aqueous solution of metformin hydrochloride， with the dose volume of 10 mL·kg^-1·d^-1. The blank group and the model group were given the same volume of distilled water. After continuous intragastric administration for 4 weeks， the body weight， body length and abdominal circumferences were measured， the Lee's obesity index was calculated， and the levels of fasting plasma glucose （FPG） and 2-hour plasma glucose （2 h PG） were detected in each group. Serum interleukin-6 （IL-6） content was detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA and protein expressions of nuclear factor kappa B （NF-<italic>κ</italic>B）， cysteinyl aspartate specific proteinase-1 （Caspase-1） and NOD-like receptor protein 3 （NLRP3） in liver tissues were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. Immunofluorescence was used to detect the expression of Caspase-1 and NLRP3 in rat liver. Hematoxylin and eosin （HE） staining was used to observe the morphology of liver tissue. Result:Compared with blank group， the body weight， abdominal circumference， body length and Lee's index in model group were significantly increased （<italic>P</italic><0.05， <italic>P</italic><0.01）， the levels of FPG and 2 h PG were significantly increased （<italic>P</italic><0.05， <italic>P</italic><0.01）， serum IL-6 content and NF-<italic>κ</italic>B， Caspase-1 and NLRP3 gene and protein expressions in liver tissue were significantly increased （<italic>P</italic><0.01）， immunofluorescence technique showed that Caspase-1 and NLRP3 expressions increased obviously in IGT rat model’s liver tissues， and inflammatory cells infiltration was observed obviously in HE stained rat liver tissue model. Compared with model group， Huanglian Wendantang and metformin hydrochloride groups could effectively reduce the body weight of IGT rats （<italic>P</italic><0.01） and abdominal obesity， and correct the levels of FPG and 2 h PG （<italic>P</italic><0.01）， while effectively reducing serum IL-6 content and gene and protein expressions of NF-<italic>κ</italic>B， Caspase-1 and NLRP3 in liver tissues of IGT rats， and alleviating inflammatory cell infiltration. Conclusion:Inflammatory response exists in IGT model rats induced by high-fat diet. Huanglian Wendantang can obviously reduce its inflammatory response and alleviate liver cell pyroptosis.

Effect of Qingre Jiedu Medicine in Treating Tumor and Regulating Signaling Pathway / 中国实验方剂学杂志

Tumor is a new organism caused by normal local tissue and cell proliferation under the effect of various tumorigenic factors. It is a serious threat to human health and life. In the theory of traditional Chinese medicine, heat poison that is similar to tumorigenic factor is one of the main causes of tumor. Modern studies have shown that Qingre Jiedu drugs have the effect in clearing heat, detoxifying, resisting bacteria and inflammation, improving immunity and resisting tumor. Therefore, Qingre Jiedu drugs are an important part of traditional Chinese prescriptions for the treatment of tumors. The research of Qingre Jiedu drugs has attracted more and more attention. Molecular pharmacological studies have shown that Qingre Jiedu drugs can play a significant role in the treatment of many kinds of tumors by regulating the targets in multiple cell signal transduction pathways. Based on literatures, due to the lack of specific review on heat and detoxification drugs for the treatment of tumor mechanism in recent years, Sophorae Flavescentis Radix, Scutellariae Radix, Hedyotis diffusa willd and Scutellariae Barbatae Herba with typical effect in clearing heat and detoxification were selected to study the tumor cell proliferation, invasion and metastasis and angiogenesis, and effects in reducing or eliminating chemotherapeutic drug resistance, inhibiting cell proliferation and inducing cell apoptosis of Sophorae Flavescentis Radix, Scutellariae Radix, Hedyotis diffusa willd and Scutellariae Barbatae Herba through Wnt/beta-catenin, phosphatidylinositol-3-kinases(PI3K)/protein kinase B(Akt), Hedgehog signaling pathways. Apparently, heat and detoxification drugs can inhibit the occurrence and development of tumors as a result of the common action of various components in heat and detoxification drugs. Meanwhile, heat and detoxification drugs have the advantages over chemical therapy and surgical treatment. This provides a reference for the application and further study of heat and detoxification drugs in tumor treatment, and new theoretical support and action target of modern medicine for the mechanism of action of traditional Chinese medicine in treating cancer.

Intervention of Epimedium 95％ethanol elution section in potential biomarkers in urine of rats based on metabonomics / 中国药理学与毒理学杂志

OBJECTIVE To observe the effect of Epimedium 95％ethanol elution section (E95EE) on endogenous metabolism in the urine of normal rats using methods of metabonomics, and to study whether differential expressions of biomarkers can influence different systems of the body along with various body-fluids-cycle distribution. METHODS SD rats were randomly divided into blank control group and E95EE group(10 rats per group). The rats of E95EE group were ig administered with E95EE 17.1 g · L-1, once daily, for 20 d, while those of normal control group were ig given an equal volume of saline. On the day of the final E95EE administration, the urine of 12 h was collected for analysis by UPLC-TOF/MS. RESULTS This study identified nine differential endogenous metabolites (3-sn-phos-phatidate, 5, 10-methenyltetrahydrofolate, l-1-phosphatidylethanolamine, 1-acyl-glycerone 3-phosphate, N-formimidoyl-l-glutamate, keto-oxaloacetate, sulfurous acid, formyl-N-acetyl-5-methoxykynurenamin and N-acetyl-5-hydroxytryptamine) and six primary metabolic pathways [glycerophospholipid metabolism, vitamin A (retinol) metabolism, histidine degradation, tryptophan metabolism, acetyl-CoA biosynthesis from citrate, glycolysis and gluconeogenesis]. The possible role of protection of E95EE discovered in the nervous and cardiovascular systems was displayed by the decreased level of 3-sn-phosphatidate and an increased level of N-formimidoyl-l-glutamate. However, the possible toxicity of E95EE on neoplastic prevention was achieved by reducing the level of l-1-phosphatidylethanolamine and 5,10-methenyl tetra-hydrofolate. CONCLUSION E95EE can produce both protection and toxicity on nervous, cardiovascular and immune systems, as well as on tumor-associated diseases. The mechanisms may be related to metabolic pathways of triglycerides, vitamin A, tryptophane and histidines.

Comparative study on serum pharmacochemistry of Gardenia jasminoides under physiological and pathological status / 中草药

Objective: By means of serum pharmacochemistry combined with modern chromatographic technology to elucidate the material basis for efficacy of aqueous extract from Gardenia jasminoides (AEGJ), which has antipyretic activities with bitter-cold property. Methods: Waters 2695 HPLC was employed as the detection system, based on the same chromatographic eluted condition, physiological and DNP-induced fever syndrome serum samples, collected from hepatic portal vein of rats after ig administration of AEGJ, were under determining to compare the absorptive variation of the chemical compounds. Results: The chromatographic fingerprints of common peaks for six batches of AEGJ were established to ascertain the optimized administration dosage, blood collection time, and the method used to process the serum. Compared with the normal administrated group under physiological status of rats, we found that the significant increment absorption of jasminoidin and the generation of two new migration compounds in the DNP fever syndrom model group. Conclusion: The increment absorption and the migration of chemical compounds in serum of DNP fever syndrome pathological model of rats might reasonably lead us to consider these chemical compounds as symbols of the bitter-cold property of G jasminoides.