The flavonoid fisetin ameliorates renal fibrosis by inhibiting SMAD3 phosphorylation, oxidative damage, and inflammation in ureteral obstructed kidney in mice

Renal fibrosis is characterized by the accumulation of extracellular matrix and inflammatory cells and kidney dysfunction, which is a major pathway in the progression of chronic kidney disease (CKD). Accumulating evidence indicates that oxidative stress plays a critical role in the initiation and progression of CKD via proinflammatory and profibrotic signaling pathways. Fisetin (3,3′,4′,7-tetrahydroxyflavone) has biological activities including antioxidant, anti-inflammatory, and anti-aging effects. Therefore, we evaluated the antifibrotic effects of fisetin on unilateral ureteral obstruction (UUO)-induced kidneys. Methods: C57BL/6 female mice were subjected to right UUO and intraperitoneally injected every other day with fisetin (25 mg/kg/ day) or vehicle from 1 hour before surgery to 7 days after surgery. Kidney samples were analyzed for renal fibrosis (α-smooth muscle actin [α-SMA] expression, collagen deposition, and transforming growth factor [TGF] β1/SMAD3 signaling pathway), oxidative damage (4-HNE and 8-OHdG expression), inflammation (proinflammatory cytokine/chemokine, macrophage, and neutrophil infiltration), and apoptosis (TUNEL staining). Cultured human proximal tubule cells were treated with fisetin before TGF-β to confirm the TGF-β downstream pathway (SMAD2/3 phosphorylation). Results: We found that fisetin treatment protected against renal fibrosis by inhibiting the phosphorylation of SMAD3, oxidative damage, inflammation, apoptotic cell death, and accumulation of profibrotic M2 macrophages in the obstructed kidneys. In cultured human proximal tubular cells, fisetin treatment inhibited TGF-β1–induced phosphorylation of SMAD3 and SMAD2. Conclusion: Fisetin alleviates kidney fibrosis to protect against UUO-induced renal fibrosis, and could be a novel therapeutic drug for obstructive nephropathy.

Shortening of primary cilia length is associated with urine concentration in the kidneys

The primary cilium, a microtubule-based cellular organelle present in certain kidney cells, functions as a mechano-sensor to monitor fluid flow in addition to various other biological functions. In kidneys, the primary cilia protrude into the tubular lumen and are directly exposed to pro-urine flow and components. However, their effects on urine concentration remain to be defined. Here, we investigated the association between primary cilia and urine concentration. Methods: Mice either had free access to water (normal water intake, NWI) or were not allowed access to water (water deprivation, WD). Some mice received tubastatin, an inhibitor of histone deacetylase 6 (HDAC6), which regulates the acetylation of α-tubulin, a core protein of microtubules. Results: WD decreased urine output and increased urine osmolality, concomitant with apical plasma membrane localization of aquaporin 2 (AQP2) in the kidney. After WD, compared with after NWI, the lengths of primary cilia in renal tubular epithelial cells were shortened and HDAC6 activity increased. WD induced deacetylation of α-tubulin without altering α-tubulin levels in the kidney. Tubastatin prevented the shortening of cilia through increasing HDAC6 activity and consequently increasing acetylated α-tubulin expression. Furthermore, tubastatin prevented the WD-induced reduction of urine output, urine osmolality increase, and apical plasma membrane localization of AQP2. Conclusions: WD shortens primary cilia length through HDAC6 activation and α-tubulin deacetylation, while HDAC6 inhibition blocks the WD-induced changes in cilia length and urine output. This suggests that cilia length alterations are involved, at least in part, in the regulation of body water balance and urine concentration.

Diminazene aceturate exacerbates renal fibrosis after unilateral ureteral obstruction in female mice

Diminazene aceturate (DIZE), an angiotensin-converting enzyme 2 (ACE2) activator, exerts anti-inflammatory and antifibrotic effects in a variety of human chronic diseases. However, the role of DIZE in kidney fibrosis and the underlying mechanism remain unclear. Therefore, we investigated the effects of DIZE on the progression of renal fibrosis after unilateral ureteral obstruction (UUO), a well-established model of chronic kidney disease. Methods: C57BL/6 female or male mice were subjected to right UUO. Mice received 15 mg/kg DIZE or vehicle (saline) daily. On the 7th day after UUO, kidneys were collected for analysis of renal fibrosis (α-smooth muscle actin, phosphorylated SMAD3, transforming growth factor (TGF)-β, Masson’s trichrome, and Sirius red staining), inflammation (macrophage infiltration, proinflammatory cytokines/ chemokines), apoptosisecrotic cell death (TUNEL and periodic acid-Schiff staining), and ACE2 activity and messenger RNA (mRNA) expression. Results: Treatment with DIZE exacerbated renal fibrosis by upregulating the profibrotic TGF-β/SMAD3 pathway, proinflammatory cytokine/chemokines (interleukin [IL]-1β, monocyte chemoattractant protein-1, IL-6, and macrophage inflammatory protein-2) levels, M2 macrophage accumulation (CD206, IL-4, IL-10, and CX3CL1), and apoptoticecrotic cell death in the obstructed kidneys of female mice but not male mice. However, DIZE treatment had no effect on ACE2 activity or mRNA expression. Conclusion: DIZE exacerbates UUO-induced renal fibrosis by aggravating tubular damage, apoptosis, and inflammation through independent of angiotensin (1–7), angiotensin II levels, and ACE2 expression/activity, rather than protecting against renal fibrosis after UUO. DIZE also has powerful effects on recruiting macrophages, including the M2-polarized subtype, in female UUO mice.

Hydrogen sulfide, a gaseous signaling molecule, elongates primary cilia on kidney tubular epithelial cells by activating extracellular signal-regulated kinase

Primary cilia on kidney tubular cells play crucial roles in maintaining structure and physiological function. Emerging evidence indicates that the absence of primary cilia, and their length, are associated with kidney diseases. The length of primary cilia in kidney tubular epithelial cells depends, at least in part, on oxidative stress and extracellular signal-regulated kinase 1/2 (ERK) activation. Hydrogen sulfide (H2S) is involved in antioxidant systems and the ERK signaling pathway. Therefore, in this study, we investigated the role of H2S in primary cilia elongation and the downstream pathway. In cultured Madin-Darby Canine Kidney cells, the length of primary cilia gradually increased up to 4 days after the cells were grown to confluent monolayers. In addition, the expression of H2S-producing enzyme increased concomitantly with primary cilia length. Treatment with NaHS, an exogenous H2S donor, accelerated the elongation of primary cilia whereas DL-propargylglycine (a cystathionine γ-lyase inhibitor) and hydroxylamine (a cystathionine-β-synthase inhibitor) delayed their elongation. NaHS treatment increased ERK activation and Sec10 and Arl13b protein expression, both of which are involved in cilia formation and elongation. Treatment with U0126, an ERK inhibitor, delayed elongation of primary cilia and blocked the effect of NaHS-mediated primary cilia elongation and Sec10 and Arl13b upregulation. Finally, we also found that H 2 S accelerated primary cilia elongation after ischemic kidney injury. These results indicate that H2S lengthens primary cilia through ERK activation and a consequent increase in Sec10 and Arl13b expression, suggesting that H2S and its downstream targets could be novel molecular targets for regulating primary cilia.

Yes-associated protein 1 as a prognostic biomarker and its correlation with telomerase in various cancers

Objectives@#The aims of this study were to investigate the expression of Yes-associated protein 1 (YAP1), its prognostic significance, and the correlation between YAP1 and telomerase in various cancers. @*Methods@#The Gene Expression Profiling Interactive Analysis database was used to analyze RNA sequencing data and the survival rate of patients with various cancers in The Cancer Genome Atlas (TCGA) database. PrognoScan was used to analyze the prognostic value of YAP1 expression in various cancers. Tumor Immune Estimation Resource was used to determine the correlation between YAP1 expression and telomerase in various cancer types based on TCGA data. @*Results@#The analysis suggested that YAP1 was differentially expressed between tissues of various cancers and non-tumor tissues. High YAP1 expression was also related to a poor prognosis in adrenocortical carcinoma, bladder urothelial carcinoma, and pancreatic adenocarcinoma. Moreover, YAP1 expression was correlated with the expression of telomerase reverse transcriptase and telomerase RNA component in various cancer types. @*Conclusion@#These results suggest that YAP1 is a potential biomarker with prognostic significance and relevance for oncogene research in various cancer types. The correlation between the expression of YAP1 and telomere-associated genes will help to understand their cancer-promoting mechanisms and interactions.

Mechanisms and therapeutic targets of ischemic acute kidney injury

Acute kidney injury (AKI) due to renal ischemia reperfusion (IR) is a major clinical problem without effective therapy and is a significant and frequent cause of morbidity and mortality during the perioperative period. Although the pathophysiology of ischemic AKI is not completely understood, several important mechanisms of renal IR-induced AKI have been studied. Renal ischemia and subsequent reperfusion injury initiates signaling cascades mediating renal cell necrosis, apoptosis, and inflammation, leading to AKI. Better understanding of the molecular and cellular pathophysiological mechanisms underlying ischemic AKI will provide more targeted approach to prevent and treat renal IR injury. In this review, we summarize important mechanisms of ischemic AKI, including renal cell death pathways and the contribution of endothelial cells, epithelial cells, and leukocytes to the inflammatory response during ischemic AKI. Additionally, we provide some updated potential therapeutic targets for the prevention or treatment of ischemic AKI, including Toll-like receptors, adenosine receptors, and peptidylarginine deiminase 4. Finally, we propose mechanisms of ischemic AKI-induced liver, intestine, and kidney dysfunction and systemic inflammation mainly mediated by Paneth cell degranulation as a potential explanation for the high mortality observed with AKI.

Periodontal health status of adolescents aged 15 years in Jinju city / 대한구강보건학회지

OBJECTIVES: The purpose of this study was to evaluate the periodontal status of adolescents aged 15 years in Jinju city. METHODS: The study subjects were 506 adolescents aged 15 years in Jinju city. We investigated the prevalence and severity of periodontal disease using the Community Periodontal Index (CPI) recommended by the WHO. Data on the frequency of daily tooth-brushing were collected through self-reported questionnaires. The information obtained on both the periodontal health status and frequency of daily toothbrushing of adolescents in Jinju city was compared with the data from the 6th Korea National Health and Nutrition Examination Survey 2013-2015 (KNHANES-VI). The software utilized in the analysis was SPSS version 23. Statistical significance was set at P < 0.05. RESULTS: The rate of healthy periodontal status in Jinju city was lower compared to KNHANES-VI (57.7% versus 63.7%). The rate of gingival bleeding in Jinju city and KNHANES-VI was 11.3% and 10.8%, respectively. The rate of calculus in Jinju city and KNHANES-VI was 31.0% and 25.6%, respectively. The rates of gingival bleeding and calculus were not significantly different between Jinju city and KNHANES-VI. The healthy periodontal segments in Jinju city were more than those in KNHANES-VI (5.43 versus 5.25). The bleeding periodontal segments in Jinju city were less than those in KNHANES-VI (0.25 versus 0.45). However, the periodontal segments with calculus in Jinju city were not significantly different from those of KNHANES-VI (0.31 versus 0.30). The frequency of daily tooth-brushing in Jinju city was more than that in KNHANES-VI (2.67 versus 2.47). CONCLUSIONS: These findings suggest that appropriate oral health education should be widely conducted to promote periodontal health in adolescents.

Onion peel water extracts enhance immune status in forced swimming rat model / 한국실험동물학회지

Onion peel contains a high concentration of quercetin and other flavonoids. In this study, the potential immune-enhancing effects of an onion peel water extract (OPE) supplement were investigated by the rat forced swimming test. OPE was prepared using hot water. Thirty-six male Sprague Dawley rats were fed a pellet diet for 1 week and were then randomly divided into six groups: normal control (NC), forced swimming control (FSC), positive control (quercetin 20 mg/kg), and three groups administered 4, 20, or 100 mg/kg of OPE. Oral drug administration was conducted daily for 4 weeks. All rats, except those of NC group, were forced to swim in water and were considered exhausted when they failed to rise to the water surface to breathe within a 7-s period. Blood lymphocyte counts, immune organ weights, histopathological analysis, and serum interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, and interleukin (IL)-12 levels were determined. OPE-treated rats consumed more food and had an increased thymic cortex to medulla ratio than that observed in FSC group rats (P<0.05). The area of the white pulp in the spleens of OPE-treated group rats was increased compared with that in FSC group rats (P<0.05). Furthermore, blood lymphocyte numbers and IFN-gamma, TNF-alpha, and IL-12 concentrations were significantly higher in OPE-fed groups than in FSC group (P<0.05). These results suggest that an OPE supplement can improve the immune status by increasing the number of immune-related cells and specific cytokine levels.

Red ginseng extract increases the functional components of silkworm Bombyx mori L. used as an anti-diabetes medicinal resource / 동물의과학연구지

The anti-diabetes mechanism of silkworm Bombyx mori L. powder and extracts was found to inhibit the activity of alpha-glycosidase. The major functional component of silkworm powder was 1-deoxynojirimycin (1-DNJ), which exerts a blood glucose-lowering effect. In this study, we aimed to compare the effects of the supplements, including red ginseng extract on the functional components of silkworm. Fifty silkworm larvae were divided into the control group (Con, N=50), group A (A, artificial diet 95% and mulberry leaf powder 5%), group B (B, artificial diet 95% and mulberry powder 5%), group C (C, artificial diet 95% and Rubus coreanus remainders 5%), group D (D, artificial diet 95% and red ginseng extract 5%), and group E (E, artificial diet 95% and yeast powder (Saccharomyces cerevisiae). Body weights and length of silkworm larvae showed significant improvement in group A, D. In particular, the growth rate in group D (artificial diet 95% and red ginseng extract 5%) was larger than that of Con. In addition, the results showed that 1-DNJ concentration was significantly largest in group D. From these results, it is concluded that the addition of red ginseng extract may be effective for larval growth and 1-DNJ accumulation in silkworm rearing with an artificial diet.

Effects of onion peel water extracts on swimming endurance in rat / 동물의과학연구지

Onion (Allium cepa L.) contains high levels of dietary fibers and antioxidants, including vitamin C, D, and folates. Onion is also known as a quercetin-rich vegetable with high flavonoid content. Onion peel contains over 20 times more quercetin than onion flesh. The aim of this study was to examine the question of whether onion peel extract supplementation has an effect on maximal exercise performance in rat. Onion peel extracts were extracted with hot water. Thirty male Sprague Dawley rats were maintained on a pellet diet for one week, and then randomly divided into five groups: Normal control, Positive control (quercetin 20 mg/kg), Onion peel 4 mg/kg, Onion peel 20 mg/kg, and Onion peel 100 mg/kg. Oral administration was performed daily. The experimental period was four weeks. Thereafter, animals were then forced to swim in water and the maximal exercise performance period from the swimming start time to the exhausted time, in which they failed to rise to the surface of the water to breathe within a 7 second period, was measured. After necropsy, weights of gastrocnemius muscles were measured. Lactate dehydrogenase concentration in serum was measured using an enzymatic method, using a commercial kit. The maximal exercise performance period was significantly longer in the onion peel extracts fed groups, compared with the control group. The lactate dehydrogenase concentration of the onion peel extracts fed groups was significantly lower, compared with the control group. Based on these results, we suggest that onion peel water extract supplementation can enhance exercise capacity caused by the mechanism of decreasing lactate dehydrogenase concentration.

Spontaneous Sertoli Cell Tumor with Cryptorchism in a Beagle Dog / 한국실험동물학회지

A male one year-old beagle dog with unilateral cryptorchism was presented for investigation of reduced appetite. Abdominal sonography and radiography demonstrated abnormal enlargement of the left testicle in the abdominal cavity. Both the retroperitoneal cryptorchid testicle and the other contralateral testicle were removed surgically. The retroperitoneal cryptorchid testicle was an enlarged, firm and bulging sphere mass. The cut surface revealed a homogeneous white color. The contralateral testicle in the scrotum showed an almost normal appearance. Histopathologically, the retroperitoneal cryptorchid testicle was diagnosed as a Sertoli cell tumor. This report describes a case of Sertoli cell tumor with cryptorchism in a beagle dog.

Comparison of Nodal Staging, of UICC TNM and Japanese Classification, and Prognostic Nodal Grouping of UICC N3M0 in Advanced Gastric Cancer

PURPOSE: We analyzed cases of advanced gastric cancer (AGC) by using two nodal stagings, UICC and Japanese systems. We also analyzed cases of UICC N3M0 by different ways to see which nodal system or group had better prognostic power. MATERIALS AND METHODS: From Feb.1990 to May 2000, 197 UICC M0 patients of AGC who had undergone curative resection were analyzed by using the nodal stagings of the UICC and the Japanese systems. Also, 58 patients with UICC N3M0 gastric cancer were analyzed by using the Japanese n-staging, metastatic ratio and the metastatic number. RESULTS: The 5-year survival rates were 62.9%, 33.0% and 21.2% for UICC N1, N2 and N3, and 61.2% and 25.3% for Japanese n1 and n2, respectively in patients of N3M0 AGC, the 5-year survival rates were 62.5% for Japanese n1, and 33.0% and 22.9% for metastatic ratios of less than 0.5 and metastatic numbers below 26, respectively significantly better than the 5-year survival rates for higher ratios and numbers (P=0.018, 0.021). CONCLUSION: UICC N staging of gastric cancer has better prognostic power with differentiation between stages than Japanese n staging. In patients with UICC N3M0 gastric cancer, the metastatic ratio and the metastatic number, as well as the Japanese n staging, were valuable prognostic factors.

The developmental expression of voltage dependent calcium channel alpha1 subunits (alpha1D, alpha1B, alpha1A, alpha1E) mRNA in the rat brain / 대한해부학회지

Voltage dependent calcium channels (VDCCs) mediate Ca++ influx into cells and are responsible for regulation of a variety of physiological effects. The key functional property of VDCCs are attributed to the calcium-pore forming alpha1 subunit. In this study, distribution pattern of alpha1 subunit (alpha1D, alpha1B, alpha1A, alpha1E) mRNA of VDCCs in developing and adult rat brain was investigated by in situ hybridization histochemistry. In the adult rat brain, each alpha1 subunit mRNA displayed a specific and distinct distribution pattern. alpha1D was highly expressed in the olfactory bulb, dentate gyrus, pituitary gland, pineal gland, hypothalamus, superior colliculus and cerebellum. Relatively low level of alpha1B was expressed throughout the whole brain and strong expression of alpha1A was observed in CA3 area of Ammon's horn, medial geniculate body, inferior colliculus and cerebellum. High level of alpha1E was found in the olfactory bulb, hippocampus, dentate gyrus, medial habenular nucleus and cerebellum. Moreover, alpha1B, alpha1A and alpha1E were expressed only in the nervous system but alpha1D was expressed not only in the nervous system but also in other tissues including liver, heart, lung and skeletal muscle. Generally the expression of alpha1D, alpha1A, and alpha1E subunit was observed from E14 and thereafter the intensity of labeling was gradually increased to P14 and then decreased to the adult level. But the expression of alpha1B subunit was observed from E14 and gradually increased to E20 and P0 and then decresaed. From the differential expressions of VDCC alpha1 subunits in developing and adult rat brain, it is suggested that each type of VDCCs may play a distinct roles in neural and nonneural tissues, and the VDCCs may be related with development of nervous system.