Synergistic effect of schizandrin A and DNase I knockdown on high glucose induced beta cell apoptosis by decreasing intracellular calcium concentration.

OBJECTIVE: To explore the synergistic effect of deoxyribonuclease I (DNase I) knockdown combined with Schizandrin A (Sch A) in protecting islet beta-cells (ß-cells) from apoptosis under high-glucose (HG) conditions. METHODS: The concentration of Sch A was detected by Cell Counting Kit-8 (CCK-8). High glucose-cultured rat insulinoma beta cell line (RIN-M5F) cells were treated with Sch A and transfected with DNase I small interfering RNA (siRNA). Cell apoptosis rate and apoptosis-related protein level were examined by flow cytometry and Western blot method respectively. In addition, Na-K-adenosine triphosphatease (Na-K-ATPase) and Ca-Mg-ATPase activity, cell membrane potential, and intracellular Ca concentration was also examined respectively. RESULTS: Our study revealed that HG stimulation can cause a significant increase in DNase I level and cell apoptosis rate. However, Sch A combined with DNase I knockdown can significantly decrease the cell apoptosis rate and apoptosis-related protein levels such as BAX ( 0.05) and Caspase-3 ( 0.01). In addition, we also found that the combination of Sch A and DNase I knockdown can dramatically increase cell membrane potential level, Na-K-ATPase, and Ca-Mg-ATPase activity. Meanwhile, intracellular Ca concentration was also found to be significantly decreased by the synergistic effect of Sch A and DNase I knockdown. CONCLUSION: Overall, our study reveals a synergistic effect of Sch A and DNase I knockdown in protecting ß-cells from HG-induced apoptosis.

Arginine metabolism regulates the pathogenesis of inflammatory bowel disease.

The pathogenesis of inflammatory bowel disease (IBD) is related to genetic susceptibility, enteric dysbiosis, and uncontrolled, chronic inflammatory responses that lead to colonic tissue damage and impaired intestinal absorption. As a consequence, patients with IBD are prone to nutrition deficits after each episode of disease resurgence. Nutritional supplementation, especially for protein components, is often implemented during the remission phase of IBD. Notably, ingested nutrients could affect the progression of IBD and the prognostic outcome of patients; therefore, they should be cautiously evaluated prior to being used for IBD intervention. Arginine (Arg) is a semi-essential amino acid required for protein synthesis and intimately associated with gut pathophysiology. To help optimize arginine-based nutritional intervention strategies, the present work summarizes that during the process of IBD, patients manifest colonic Arg deficiency and the turbulence of Arg metabolic pathways. The roles of Arg-nitric oxide (catalyzed by inducible nitric oxide synthase) and Arg-urea (catalyzed by arginases) pathways in IBD are debatable; the Arg-polyamine and Arg-creatine pathways are mainly protective. Overall, supplementation with Arg is a promising therapeutic strategy for IBD; however, the dosage of Arg may need to be carefully tailored for different individuals at different disease stages. Additionally, the combination of Arg supplementation with inhibitors of Arg metabolic pathways as well as other treatment options is worthy of further exploration.

Mechanism of hepatotoxicity induced by ethanol extract of Dysosma versipellis based on "quantity-weight-evidence" network toxicology / 中国中药杂志

In this study, the toxicological/pharmacological research method of "quantity-weight-evidence" network was first proposed and practiced to supplement the existing methodology of network toxicology. We transformed the traditional qualitative network into a quantitative network in this study by attributing weights to toxic component content and target frequency, which improved the reliability of data and provided a research idea for the systematic safety evaluation and toxicological research of Chinese medicinal herbs. Firstly, 50% ethanol extract of Dysosma versipellis(DV) was administrated to rats via gavage and the potential hepatotoxic components were identified by serum pharmacochemistry. Then, the component targets were obtained from SwissTargetPrediction, PharmMapper and other online databases, and the target weights were given according to the relative content of components and target fishing frequency. Meanwhile, the targets of hepatotoxicity were predicted from online databases such as Comparative Toxicology Database(CTD) and GeneCards. Subsequently, protein-protein interaction analysis and KEGG pathway enrichment were performed with the STRING database. Finally, the quantitative network of "toxic components-weighted targets-pathways" was constructed. Eleven potential toxic compounds were predicted, including podophyllotoxin, podophyllotoxone, deoxypodophyllotoxin, and 6-methoxypodophyllotoxin. A total of 106 hepatotoxic targets and 65 weighted targets(e.g., Cdk2, Egfr, and Cyp2 c9) were identified. The results of Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment showed that these targets could act on PI3 K-AKT, MAPK, and Ras signaling pathways to play a role in inflammatory response and oxidative stress. However, traditional network toxicology showed that 51 targets such as AKT1, Alb, and Stat3 may lead to hepatotoxicity by mediating inflammation and cell proliferation. In conclusion, we proposed "quantity-weight-evidence" network toxicology in this study and used it to study the mechanism of DV-induced hepatotoxicity in rats. This study confirms the feasibility of this new methodology in toxicological evaluation and further improves the systematic evaluation of the safety of Chinese medicinal herbs.

Chondroitin Sulfate Alleviates Diabetic Osteoporosis and Repairs Bone Microstructure via Anti-Oxidation, Anti-Inflammation, and Regulating Bone Metabolism.

Diabetic osteoporosis (DOP) belongs to secondary osteoporosis caused by diabetes; it has the characteristics of high morbidity and high disability. In the present study, we constructed a type 1 diabetic rat model and administered chondroitin sulfate (200 mg/kg) for 10 weeks to observe the preventive effect of chondroitin sulfate on the bone loss of diabetic rats. The results showed that chondroitin sulfate can reduce blood glucose and relieve symptoms of diabetic rats; in addition, it can significantly increase the bone mineral density, improve bone microstructure, and reduce bone marrow adipocyte number in diabetic rats; after 10 weeks of chondroitin sulfate administration, the SOD activity level was upregulated, as well as CAT levels, indicating that chondroitin sulfate can alleviate oxidative stress in diabetic rats. Chondroitin sulfate was also found to reduce the level of serum inflammatory cytokines (TNF-α, IL-1, IL-6, and MCP-1) and alleviate the inflammation in diabetic rats; bone metabolism marker detection results showed that chondroitin sulfate can reduce bone turnover in diabetic rats (decreased RANKL, CTX-1, ALP, and TRACP 5b levels were observed after 10 weeks of chondroitin sulfate administration). At the same time, the bone OPG and RUNX 2 expression levels were higher after chondroitin sulfate treatment, the bone RANKL expression was lowered, and the OPG/RANKL ratio was upregulated. All of the above indicated that chondroitin sulfate could prevent STZ-induced DOP and repair bone microstructure; the main mechanism was through anti-oxidation, anti-inflammatory, and regulating bone metabolism. Chondroitin sulfate could be used to develop anti-DOP functional foods and diet interventions for diabetes.

Clinical Effect of Jinlida and Tongxinluo on Type 2 Diabetic Kidney Disease Under Vessel Collateral Theory / 中国实验方剂学杂志

Objective: To observe the clinical efficacy of Jinlida (JLD) granules and Tongxinluo (TXL) capsules on type 2 diabetic kidney disease (DKD) under the guidance of vessel collateral theory. Method: A total of 120 patients with type 2 DKD, were randomly divided into 2 groups:the normal control group (60 cases) and the treatment group (60 cases). The patients in normal control group were treated with dietary control and hyperglycemia control. Based on treatment in control group, patients in treatment group were additionally treated with JLD granules (1 bag, tid), and TXL capsules (4 capsules, tid). The treatment was lasted for 12 weeks. The clinical efficacy, traditional Chinese medicine(TCM) syndrome scores were observed and compared before and after treatment. At the same time, the levels of glucose metabolism indexes including fasting blood glucose (FBG),postprandial 2 h plasma glucose (2 hPG), glycosylated hemoglobin (HbA1c) and the insulin resistance (IR); the levels of lipid metabolism indexes including total cholesterol(TC),triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and the high-density lipoprotein cholesterol (HDL-C); the levels of renal function indexes including urinary albumin excretion rate (UAER) and serum creatinine (SCr); as well as nailfold microcirculation were detected and compared. Result: ①The total effective rate was 80.0% in treatment group, significantly higher than 61.67% in the normal control group (PPPPPβ2-MG in treatment group was significantly obvious than that in the control group (PPConclusion: Tongxinluo combined with Jinlida can improve the clinical symptoms of patients with type 2 diabetic nephropathy and reduce urinary trace albumin, and its mechanism may be related to lowering glucose, regulating lipid metabolism and improving microcirculation.

Effect of hyperbaric oxygen treatment on postoperative cognitive dysfunction in rats / 中华麻醉学杂志

Objective To evaluate the effect of hyperbaric oxygen (HBO) treatment on postoperative cognitive dysfunction (POCD) in rats.Methods Eighteen healthy male Sprague-Dawley rats,weighing 260-290 g,were anesthetized with intraperitoneal 10％ chloral hydrate 350 mg/kg.POCD was induced by injecting Aβ-40 2μl into the bilateral hippocampi by using a brain stereotaxic apparatus.The rats were randomly divided into 3 groups (n =6 each) using a random number table:normal sahne group (group NS),POCD group,and HBO treatment group(groupHBO).0.9％ normal saline 2 μl was injected into hippocampus in group NS.In group POCD,Aβ0 2 μl was injected into hippocampus.In group HBO,Aβ 2μl was injected into hippocampus,and then the rats received hyperbaric oxygen treatment lasting for 60 min once a day within 1-5 days after operation.Morris water maze test was performed on 7,14 and 21 days after operation in each group and the swimming distance and speed and escape latency were recorded.The animals were sacrificed after the end of test,the hippocampi were then removed to detect the activation of astrocytes (by immuno-histochemistry) and content of tumor necrosis factor-α (TNF-α) (by ELISA).Resets There were no significant differences in the parameters of behavior in Morris water maze test on 7 and 14 days after operation between the three groups.Compared with group NS,the swimming distance and escape latency were significantly prolonged,and the activation of astrocytes and TNF-α content were increased on 21 days after operation in group POCD,and the swimming distance and escape latency were significantly prolonged,the activation of astrocytes was increased,and no significant change was found in TNF-α content on 21 days after operation in group HBO.Compared with group POCD,the swimming distance and escape latency were significantly shortened,and the activation of astrocytes and TNF-α content were decreased in group HBO.There was no significant difference in the swimming speed at each time point among the three groups.Conclusion HBO treatment can alleviate POCD in rats,and the mechanism is related to inhibition of activation of astrocytes and inflammatory responses in hippocampi by HBO.

Activation of peroxisome proliferator-activated receptor-alpha protects the heart from ischemia/reperfusion injury.

BACKGROUND: Peroxisome proliferator-activated receptor-alpha (PPAR-alpha) is expressed in the heart and regulates genes involved in myocardial fatty acid oxidation (FAO). The role of PPAR-alpha in acute ischemia/reperfusion myocardial injury remains unclear. METHODS AND RESULTS: The coronary arteries of male mice were ligated for 30 minutes. After reperfusion for 24 hours, ischemic and infarct sizes were determined. A highly selective and potent PPAR-alpha agonist, GW7647, was administered by mouth for 2 days, and the third dose was given 1 hour before ischemia. GW7647 at 1 and 3 mg x kg(-1) x d(-1) reduced infarct size by 28% and 35%, respectively (P<0.01), and myocardial contractile dysfunction was also improved. Cardioprotection by GW7647 was completely abolished in PPAR-alpha-null mice. Ischemia/reperfusion downregulated mRNA expression of cardiac PPAR-alpha and FAO enzyme genes, decreased myocardial FAO enzyme activity and in vivo cardiac fat oxidation, and increased serum levels of free fatty acids. All of these changes were reversed by GW7647. Moreover, GW7647 attenuated ischemia/reperfusion-induced release of multiple proinflammatory cytokines and inhibited neutrophil accumulation and myocardial expression of matrix metalloproteinases-9 and -2. Furthermore, GW7647 inhibited nuclear factor-kappaB activation in the heart, accompanied by enhanced levels of inhibitor-kappaBalpha. CONCLUSIONS: Activation of PPAR-alpha protected the heart from reperfusion injury. This cardioprotection might be mediated through metabolic and antiinflammatory mechanisms. This novel effect of the PPAR-alpha agonist could provide an added benefit to patients treated with PPAR-alpha activators for dyslipidemia.