Hypoxia-inducible factor-1α activation can attenuate renal podocyte injury and alleviate proteinuria in rats in a simulated high-altitude environment.

OBJECTIVE: The aims of this study were to understand whether podocyte injury is involved in proteinuria after rapid ascent to high altitude and to explore whether hypoxia-inducible factor (HIF)-1α is involved in the adaptive regulation of this proteinuria. METHODS: Rats in the experimental group were housed in a low-pressure oxygen chamber to simulate a high-altitude environment (5,000 m). The intervention group was placed under the same conditions as the experimental group and prolyl-hydroxylase inhibitor (PHI) was intraperitoneally injected. The control group was housed in a low altitude environment (500 m). On days 0, 7, 14, and 28, urinary albumin quantification and electrophoresis were performed. The expression levels of CD2-associated protein (CD2AP), nephrin and HIF-1α were detected by immunofluorescence. RESULTS: The medium and large molecule proteins with molecular weights ranging from 63 to 75 kD were present in the urine of rats in the experimental group and that the urinary albumin levels first increased and then decreased with time and the increase on day 14 was most significant (24.58 ± 4.30 mg on day 14 VS 5.13 ± 1.58 mg on day 0). Electron microscopy revealed podocyte lesions in rats in the experimental group. Immunofluorescence results showed that the protein expression levels of CD2AP and nephrin in the glomeruli of rats in the experimental group were lower than those in the control group (P < 0.001) and that the expression levels of which in the intervention group were higher than those in the experimental group (P < 0.001). The expression of HIF-1α protein in the renal tissues of rats in the experimental group was higher than that in the control group (P < 0.001) and lower than that in the intervention group (P < 0.001). CONCLUSION: The podocyte injury may be involved in the occurrence of proteinuria after rapid ascent to high altitude. PHI may have a potential role in reducing proteinuria by upregulating local HIF-1α expression in the kidney to alleviate podocyte injury.

Angelica sinensis Polysaccharide and Astragalus membranaceus Polysaccharide Accelerate Liver Regeneration by Enhanced Glycolysis via Activation of JAK2/STAT3/HK2 Pathway.

The promotion of liver regeneration is crucial to avoid liver failure after hepatectomy. Angelica sinensis polysaccharide (ASP) and Astragalus membranaceus polysaccharide (AMP) have been identified as being associated with hepatoprotective effects. However, their roles and specific mechanisms in liver regeneration remain to be elucidated. In the present study, it suggested that the respective use of ASP or AMP strikingly promoted hepatocyte proliferation in vitro with a wide range of concentrations (from 12.5 µg/mL to 3200 µg/mL), and a stronger promoting effect was observed in combined interventions. A significantly enhanced liver/body weight ratio (4.20%) on day 7 and reduced serum transaminase (ALT 243.53 IU/L and AST 423.74 IU/L) and total bilirubin (52.61 IU/L) levels on day 3 were achieved by means of ASP-AMP administration after partial hepatectomy in mice. Metabonomics showed that differential metabolites were enriched in glycolysis with high expression of beta-d-fructose 6-phosphate and lactate, followed by significantly strengthened lactate secretion in the supernatant (0.54) and serum (0.43) normalized to control. Upon ASP-AMP treatment, the knockdown of hexokinase 2 (HK2) or inhibited glycolysis caused by 2-deoxy-d-glucose decreased hepatocyte proliferation in vitro and in vivo. Furthermore, pathway analysis predicted the role of JAK2/STAT3 pathway in ASP-AMP-regulated liver regeneration, and phosphorylation of JAK2 and STAT3 was proven to be elevated in this promoting process. Finally, downregulated expression of HK2, an attenuated level of lactate secretion, and reduced hepatocyte proliferation were displayed when STAT3 was knocked out in vitro. Therefore, it can be concluded that ASP-AMP accelerated liver regeneration and exerted a hepatoprotective effect after hepatectomy, in which the JAK2/STAT3/HK2 pathway was actively involved in activating glycolysis.

[Dexmedetomidine suppresses mechanical allodynia by inhibiting hyperpolarization-activated inward current].

The purpose of the present study was to investigate the effects of dexmedetomidine (DEX) on neuropathic pain in the chronic compression of dorsal root ganglion (CCD) rat model and the underlying mechanism. Pain behavioral tests were applied to observe the effects of DEX on mechanical allodynia in Sprague Dawley (SD) rats. Whole cell patch clamp was used to observe the influence of DEX on excitability and hyperpolarization-activated inward current (Ih) of C- and Aδ-type dorsal root ganglion (DRG) neurons. The results showed that mechanical allodynia of CCD rats was significantly inhibited by DEX (P < 0.05). In C- and Aδ-type DRG neurons from the CCD rats, DEX significantly increased rheobase and after hyperpolarizing potential, as well as decreased Ih current density. These results suggest that DEX could attenuate the neuropathic pain in the CCD rats, and the mechanism might be related to the depressed Ih current density and excitability of C- and Aδ-type DRG neurons.

Dispelling Dampness, Relieving Turbidity and Dredging Collaterals Decoction, Attenuates Potassium Oxonate-Induced Hyperuricemia in Rat Models.

Purpose: Dispelling dampness, relieving turbidity and dredging collaterals decoction (DED), is a traditional Chinese medicine used in the treatment of hyperuricemia. We aimed to explore the effect and mechanism of DED in the treatment of hyperuricemia. Methods: The effects of DED (9.48, 4.74, and 2.37 g/kg/d) on potassium oxonate (750 mg/kg/d)-induced hyperuricemia in rats were evaluated by serum uric acid (UA), creatinine (CRE), blood urea nitrogen (BUN), and renal pathological changes. Network pharmacology was used to identify the effective components and targets of DED, and the key targets and signaling pathways for its effects on hyperuricemia were screened. Molecular docking was used to predict the action of DED. H&E, immunohistochemistry, WB, and PCR were used to validate the network pharmacology results. Results: DED can effectively alleviate hyperuricemia, inhibit UA, CRE, BUN, and xanthine oxidase (XOD) activity, and reduce renal inflammatory cell infiltration and glomerular atrophy. The experiment identified 27 potential targets of DED for hyperuricemia, involving 9 components: wogonin, stigmasterol 3-O-beta-D-glucopyranoside, 3ß-acetoxyatractylone, beta-sitosterol, stigmasterol, diosgenin, naringenin, astilbin, and quercetin. DED can relieve hyperuricemia mainly by inhibiting RAGE, HMGB1, IL17R, and phospho-TAK1, and by regulating the AGE-RAGE and IL-17 signaling pathways. Conclusion: DED can alleviate hyperuricemia by inhibiting XOD activity and suppressing renal cell apoptosis and inflammation via the AGE-RAGE signaling pathway and IL-17 signaling pathway. This study provides a theoretical basis for the clinical application of DED.

Progesterone suppresses the progression of colonic carcinoma by increasing the activity of the GADD45α/JNK/c­Jun signalling pathway.

Colorectal cancer (CRC) is the third most diagnosed cancer worldwide. Progesterone is associated with a decreased risk of CRC and leads to a favourable prognosis. However, the specific mechanism by which progesterone suppresses malignant progression remains to be elucidated. In the present study, the level of progesterone was first analysed in 77 patients with CRC, and immunohistochemistry was performed to detect the expression of progesterone receptor (PGR) in the paired specimens. The correlations between progesterone, PGR and CRC prognosis were assessed. A Cell Counting Kit­8 assay was then used to detect proliferation of the CRC cells. Flow cytometry was performed to estimate apoptosis and to evaluate the cycle of the CRC cells. A xenograft tumour model was established in nude mice to assess the role of progesterone in tumour growth. Finally, a PCR microarray was used to screen differentially expressed genes to further interpret the mechanism by which progesterone inhibits the malignant progression of CRC. It was found that low expression of progesterone and PGR were significantly associated with poor prognosis of CRC. In addition, progesterone suppressed CRC cell proliferation by arresting the cell cycle and inducing apoptosis in vitro. Moreover, the inhibitory role of progesterone in tumour growth was verified in vivo. Further investigation showed that the level of growth arrest and DNA damage­inducible protein α (GADD45α) was up­regulated by progesterone, and this was followed by the activation of the JNK pathway. Progesterone increased the activity of the JNK pathway via GADD45α to inhibit proliferation by arresting the cell cycle and inducing apoptosis, thereby suppressing the malignant progression of CRC. Therefore, it can be concluded that progesterone and PGR might act as inhibiting factors for poor prognosis of CRC.

Abdominal paracentesis drainage attenuates intestinal inflammation in rats with severe acute pancreatitis by inhibiting the HMGB1-mediated TLR4 signaling pathway.

BACKGROUND: Our previous studies confirmed that abdominal paracentesis drainage (APD) attenuates intestinal mucosal injury in rats with severe acute pancreatitis (SAP), and improves administration of enteral nutrition in patients with acute pancreatitis (AP). However, the underlying mechanisms of the beneficial effects of APD remain poorly understood. AIM: To evaluate the effect of APD on intestinal inflammation and accompanying apoptosis induced by SAP in rats, and its potential mechanisms. METHODS: SAP was induced in male adult Sprague-Dawley rats by 5% sodium taurocholate. Mild AP was induced by intraperitoneal injections of cerulein (20 µg/kg body weight, six consecutive injections). Following SAP induction, a drainage tube connected to a vacuum ball was placed into the lower right abdomen of the rats to build APD. Morphological changes, serum inflammatory mediators, serum and ascites high mobility group box protein 1 (HMGB1), intestinal barrier function indices, apoptosis and associated proteins, and toll-like receptor 4 (TLR4) signaling molecules in intestinal tissue were assessed. RESULTS: APD significantly alleviated intestinal mucosal injury induced by SAP, as demonstrated by decreased pathological scores, serum levels of D-lactate, diamine oxidase and endotoxin. APD reduced intestinal inflammation and accompanying apoptosis of mucosal cells, and normalized the expression of apoptosis-associated proteins in intestinal tissues. APD significantly suppressed activation of the intestinal TLR4 signaling pathway mediated by HMGB1, thus exerting protective effects against SAP-associated intestinal injury. CONCLUSION: APD improved intestinal barrier function, intestinal inflammatory response and accompanying mucosal cell apoptosis in SAP rats. The beneficial effects are potentially due to inhibition of HMGB1-mediated TLR4 signaling.