Targeted Restoration of GPX3 Attenuates Renal Ischemia/Reperfusion Injury by Balancing Selenoprotein Expression and Inhibiting ROS-mediated Mitochondrial Apoptosis.

BACKGROUND: Renal ischemia/reperfusion (IR) injury is the leading cause of acute kidney injury in both autologous and transplanted kidneys. Low-level glutathione peroxidase 3 (GPX3) is associated with renal IR injury. The exact mechanism of targeted GPX3 restoration in renal IR injury has yet to be determined. METHODS: The distribution of GPX3 in different tissues and organs of the body was investigated. The level of GPX3 in renal IR injury was assessed. To confirm the action of GPX3 and its mechanisms, IR models were used to introduce adeno-associated virus 9 containing GPX3, as well as hypoxia/reoxygenation-exposed normal rat kidney cells that consistently overexpressed GPX3. Reverse molecular docking was used to confirm whether GPX3 was a target of ebselen. RESULTS: GPX3 is abundant in the kidneys and decreases in expression during renal IR injury. GPX3 overexpression reduced renal IR injury and protected tubular epithelial cells from apoptosis. Proteomics analysis revealed a strong link between GPX3 and mitochondrial signaling, cellular redox state, and different expression patterns of selenoproteins. GPX3 inhibited reactive oxygen species-induced mitochondrial apoptosis and balanced the disordered expression of selenoproteins. GPX3 was identified as a stable selenoprotein that interacts with ebselen. Ebselen enhanced the level of GPX3 and reduced IR-induced mitochondrial damage and renal dysfunction. CONCLUSIONS: Targeted restoration of GPX3 attenuates renal IR injury by balancing selenoprotein expression and inhibiting reactive oxygen species-mediated mitochondrial apoptosis, indicating that GPX3 could be a potential therapeutic target for renal IR injury.

Validation of CSF-1 receptor (CD115) staining for analysis of murine monocytes by flow cytometry.

CD115, the receptor for colony stimulating factor 1, is essential for survival and differentiation of monocytes and macrophages and is therefore frequently used to define monocyte subsets and their progenitors in immunological assays. However, CD115 surface expression and detection by flow cytometry is greatly influenced by cell isolation and processing methods, organ source, and disease context. In a systematic analysis of murine monocytes, we define experimental conditions that preserve or limit CD115 surface expression and staining by flow cytometry. We also find that, independent of conditions, CD115 surface levels are consistently lower in Ly6Clo monocytes than in Ly6Chi monocytes, with the exception of Ly6Clo monocytes in the bone marrow. Furthermore, in contrast to IL-34, the presence of colony stimulating factor 1 impairs CD115 antibody staining in a dose-dependent manner, which, in a model of ischemic kidney injury with elevated levels of colony stimulating factor 1, influenced quantification of kidney monocytes. Thus, staining and experimental conditions affect quantitative and qualitative analysis of monocytes and may influence experimental conclusions.

Selenoprotein Gene mRNA Expression Evaluation During Renal Ischemia-Reperfusion Injury in Rats and Ebselen Intervention Effects.

Effects of selenoproteins on many renal diseases have been reported. However, their role in renal ischemia-reperfusion (I/R) injury is unclear. The present study was performed to investigate the impact of ebselen and renal I/R injury on the expression of selenoproteins. Sprague-Dawley rats were pretreated with or without ebselen (10 mg/kg) through a daily single oral administration from 3 days before renal I/R surgery. RT-qPCR (real-time quantitative PCR) was performed to determine the mRNA expression of 25 selenoprotein genes in the renal tissues. The expression levels of two selenoproteins, including GPX3 (glutathione peroxidase 3) and DIO1 (iodothyronine deiodinase 1), were evaluated by Western blot or/and IHF (immunohistofluorescence) assays. Furthermore, renal function, renal damage, oxidative stress, and apoptosis were assessed. The results showed that in renal I/R injury, the mRNA levels of 15 selenoprotein genes (GPX1, GPX3, GPX4, DIO1, DIO2, TXNRD2, TXNRD3, SEPHS2, MSRB1, SELENOF, SELENOK, SELENOO, SELENOP, SELENOS, and SELENOT) were decreased, whereas those of eight selenoprotein genes (GPX2, GPX6, DIO3, TXNRD1, SELENOH, SELENOM, SELENOV, and SELENOW) were increased. I/R also induced a reduction in the expression levels of GPX3 and DIO1 proteins. In addition, our results indicated that ebselen reversed the changes in those selenoprotein genes, excluding SELENOH, SELENOM, SELENOP, and SELENOT, in renal I/R injury and alleviated I/R-induced renal dysfunction, tissue damage, oxidative stress, and apoptosis. To our knowledge, this is the first study to investigate the changes of 25 mammalian selenoprotein genes in renal I/R injury kidneys. The present study also provided more evidence for the roles of ebselen against renal I/R injury.

Ebselen ameliorates renal ischemia-reperfusion injury via enhancing autophagy in rats.

Renal ischemia-reperfusion (I/R) injury is one of the most common causes of chronic kidney disease (CKD). It brings unfavorable outcomes to the patients and leads to a considerable socioeconomic burden. The study of renal I/R injury is still one of the hot topics in the medical field. Ebselen is an organic selenide that attenuates I/R injury in various organs. However, its effect and related mechanism underlying renal I/R injury remains unclear. In this study, we established a rat model of renal I/R injury to study the preventive effect of ebselen on renal I/R injury and further explore the potential mechanism of its action. We found that ebselen pretreatment reduced renal dysfunction and tissue damage caused by renal I/R. In addition, ebselen enhanced autophagy and inhibited oxidative stress. Additionally, ebselen pretreatment activated the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway. The protective effect of ebselen was suppressed by autophagy inhibitor wortmannin. In conclusion, ebselen could ameliorate renal I/R injury, probably by enhancing autophagy, activating the Nrf2 signaling pathway, and reducing oxidative stress.

[Establishment of Rat Prostatitis Model through Induction with Estrogen and Androgen at Different Concentrations].

OBJECTIVE: To establish an experimental prostatitis animal model in Sprague-Dawley (SD) rats through induction by treatment of estrogen and androgen at different concentrations. METHODS: Fifty-three male SD rats aged 3 to 4 months were used in the study, and the castration model of male rats was established by excision of bilateral testes. The rats were randomly assigned to a blank group, a castration group and treatment groups receiving estrogen and androgen at different concentrations after castration, with 4 rats in each group. Dihydrotestosterone (DHT) and estradiol (E) were administered daily by subcutaneous injection to the treatment groups. All the rats were sacrificed by way of cervical dislocation after 1 month and the serum DHT and E concentrations of the rats in each group were assessed with ELISA. Prostate specimens were collected and the relative weight of the prostate of each group of rats was calculated. After HE staining of the prostate tissue, we observed with optic microscope structural changes in the prostate tissue and the state of prostatic inflammation in each group. Immunohistochemical examination was done to assess the expression of three inflammatory factors, transforming growth factor-ß1 (TGF-ß1), interleukin (IL)-6 and IL-8, in rat prostate tissues. RESULTS: The results of HE staining of rat prostate tissue showed that, compared with the blank group and castration group, the degree of inflammation increased significantly in the E0.05+DHT 0.5 mg/kg group and DHT0.15+E0.15 mg/kg group ( P<0.05). However, once the concentration of DHT exceeded 0.5 mg/kg, the degree of inflammation did not further aggravate. The results of immunohistochemical staining showed that when the concentration of exogenous E was constant, the expression of TGF-ß1 and IL-8 increased significantly in the E0.05+DHT 0.15 mg/kg group, E0.05+DHT 0.5 mg/kg group and E0.05+DHT 1.5 mg/kg group compared with that of the blank group ( P<0.05). In the E0.05+DHT 0.15 mg/kg group and E0.05+DHT 0.5 mg/kg group, the expression of TGF-ß1 and IL-8 increased significantly compared with that of the castration group ( P<0.05). Once the concentration of DHT reached 0.5 mg/kg, further increase in the concentration of DHT did not lead to any significant changes in the expression of TGF-ß1 or IL-8. In addition, when the concentration of exogenous DHT remained unchanged, the expressions of TGF-ß1, IL-6, and IL-8 increased significantly in the DHT0.15+E 0.05 mg/kg group and DHT0.15+E 0.5 mg/kg group, compared with that of the blank group and castration group ( P<0.05). CONCLUSION: Castration combined with treatment of different concentrations of estrogen and androgen could successfully induce the prostatitis model in SD rats.

Segmental artery clamping versus main renal artery clamping in nephron-sparing surgery: updated meta-analysis.

OBJECTIVES: Ischemia-reperfusion injury is harmful in partial nephrectomy (PN) in renal cell carcinoma. Choosing an appropriate surgical method is important to reduce ischemia-reperfusion injury. This study aimed to compare the effect of segmental artery clamping (SAC) and main renal artery clamping (MAC) on patients who underwent PN. METHODS: Studies from January 2008 to November 2019 were identified by an electronic search of English and Chinese databases, including PubMed, Excerpt Medica Database, Cochrane Library, Wanfang, VIP, and Chinese National Knowledge Internet, without language restriction. Two reviewers were involved in the trial. The effects on operation time (OT), warm ischemia time (WIT), length of hospital stay (LOS), blood transfusion rate, postoperative complication rate, Clavien classification (≥ 3), and positive surgery margin (PSM) were evaluated using Stata software. Standardized mean difference (SMD, for continuous data) and pooled odds ratios (for count data) with 95% confidence interval (CI) were used as effect indicators. RESULTS: Thirty-two studies were included. SAC decreased the 1-week (SMD = - 0.973; 95% CI = - 1.414, - 0.532; P = 0.000), 1-month (SMD = - 0.411; 95% CI = - 0.769, - 0.053; P = 0.025), and 3-month (affected kidney: SMD = - 0.914; 95% CI = - 1.662, - 0.617; P = 0.000) percentages of postoperative changes in renal function (estimated glomerular filtration rate) between the SAC and MAC groups. Sub-group analysis showed that the SAC group had longer OT (SMD = 0.562; 95% CI = 0.252, 0.871; P = 0.000) than the MAC group. However, no differences were observed in the OT, WIT, LOS, blood transfusion rate, postoperative complication rate, Clavien classification (≥ 3), and PSM between the two groups. CONCLUSIONS: SAC is superior to MAC in terms of short-term postoperative renal function recovery. The use of SAC or MAC depends on tumor size, location, surgical modality, and surgeon's judgments.

Normobaric hyperoxia plays a protective role against renal ischemia-reperfusion injury by activating the Nrf2/HO-1 signaling pathway.

Following renal ischemia-reperfusion injury (RIRI), because of the decrease in oxygen supply to the kidney, a large amount of oxygen-free radicals is generated, and in severe cases, tissue cells will undergo apoptosis or even die. Normobaric hyperoxia (NBHO) is a very common clinical adjuvant treatment. It restores the oxygen supply after renal ischemia and combats oxidative stress in tissues, thus playing a protective role. In this study, our aim is to elucidate the protective mechanism of NBHO inhalation in a rat RIRI model. We performed a surgical excision of the left kidney of the rat and established a right kidney solitary kidney model. Later, the right renal pedicle of the rat was clamped using a non-invasive vascular clamp for 45 min. After the vascular clamp was released and reperfused for 24 h, the rat was placed in a closed oxygen chamber. It was subjected to inhalation of high-concentration oxygen (50%-55%), 2 h daily, for 7 days.RIRI induces postoperative weight loss, impaired renal function, increased oxygen free radicals, reduced antioxidant substances, increased histopathological damage, and increased levels of apoptosis. These effects were significantly improved after treatment with NBHO. At the same time, NBHO significantly increased the expression levels of Nrf2 and HO-1 in the tissues after RIRI. To verify whether HO-1 induced by Nrf2 is involved in the resistance to oxidative stress, after the rat RIRI and before inhaling NBHO, we intraperitoneally injected HO-1 specific inhibitor zinc protoporphyrin (ZnPP) (45 µmol/Kg). However, we found that ZnPP reversed the protective effect of NBHO on RIRI in rats. Combining all the results, we have demonstrated the protective effect of NBHO on RIRI, which can be at least partially attributed to the activation of the Nrf2/HO-1 antioxidative stress pathway.

The Prognostic Significance of EIF3C Gene during the Tumorigenesis of Prostate Cancer.

Prostate cancer (PCa) is the most common malignant tumor for men. But the mechanism is unclear. EIF3C was shown to be overexpressed in PCa tissues and cell lines. EIF3C overexpression was correlated to age and tumor stage in PCa patients and indicated poor survival. The proliferation, migration, and invasiveness of PC3 cells were all inhibited after EIF3C knockdown. Additionally, the phosphorylation level of PI3K and Akt was downregulated while total NF-κB and Myc decreased after EIF3C knockdown. But the expression of IκB increased reversely. Therefore, EIF3C at least partially regulates the activity of PI3K/Akt/NF-κB signaling pathway in PC3 cells.

Risk factors of multidrug-resistant tuberculosis in China: A meta-analysis.

BACKGROUND: Multidrug-resistant tuberculosis (MDR-TB) brings major challenges to the health care workers (HCWs). This study is to determine the risk factors for MDR-TB, latent tuberculosis infection (LTBI), and tuberculosis (TB) disease among HCWs in China. METHODS: A meta-analysis was conducted to evaluate the risk factors for MDR-TB, LTBI, and TB disease among HCWs using a random-effects model, and the pooled odds ratios (ORs) with 95% confidence interval (CI) were used as effect indicators. RESULTS: We identified 46 eligible studies and found eight factors were associated with MDR. The ORs with 95% CI are migrant population 1.96 (95% CI, 1.50-2.57), low family income 2.23 (95% CI, 1.74-2.85), retreatment 7.22 (95% CI, 5.63-9.26), anti-TB treatment history 5.65 (95% CI, 4.80-6.65), multiple episodes of treatment 3.28 (95% CI, 2.60-4.13), adverse reactions 3.48 (95% CI, 2.54-4.76), interrupted treatment 3.18 (95% CI, 2.60-3.89), and lung cavities 1.42 (95% CI, 1.14-1.77). Work duration as a HCW for 5 years and above increased the risk of LTBI and TB. HCWs aged 30 years and above were more susceptible to TB (OR = 1.70, 95% CI: 1.37-2.09). CONCLUSION: The risk factors for MDR-TB in China are possibly migrant population, low family income, retreatment, anti-TB treatment history, adverse reactions, interrupted treatment, and lung cavities. Longer work duration and greater age are risk factors for LTBI and TB among HCWs.

Comparative study of the binding mode between cytochrome P450 17A1 and prostate cancer drugs in the absence of haem iron.

According to the X-ray crystal structures of CYP17A1 (including its complexes with inhibitors), it is shown that a hydrogen bond exists between CYP17A1 and its inhibitors (such as abiraterone and TOK-001). Previous short MD simulations (50 ns) suggested that the binding of abiraterone to CYP17A1 is stronger than that of TOK-001. In this work, by carrying out long atomistic MD simulations (200 ns) of CYP17A1 and its complexes with abiraterone and TOK-001, we observed a binding mode between CYP17A1 and abiraterone, which is different from the binding mode between CYP17A1 and TOK-001. In the case of abiraterone binding, the unfilled volume in the active site cavity increases the freedom of movement of abiraterone within CYP17A1, leading to the collective motions of the helices G and B' as well as the breaking of hydrogen bond existing between the 3ß-OH group of abiraterone and N202 of CYP17A1. However, the unfilled volume in the active site cavity can be occupied by the benzimidazole ring of TOK-001, restraining the motion of TOK-001. By pulling the two inhibitors (abiraterone and TOK-001) out of the binding pocket in CYP17A1, we discovered that abiraterone and TOK-001 were moved from their binding sites to the surface of protein similarly through the channels formed by the helices G and B'. In addition, based on the free energy calculations, one can see that it is energetically favorable for the two inhibitors (abiraterone and TOK-001) to enter into the binding pocket in CYP17A1.