Corn oligopeptides inhibit Akt/NF-κB signaling pathway and inflammatory factors to ameliorate CCl4 -induced hepatic fibrosis in mice.

In this study, the effect of corn oligopeptides (COPs) with liver protection activity on mice with hepatic fibrosis (HF) induced by carbon tetrachloride (CCl4 ) was studied. It was proved that COPs can ameliorate the liver injury and inflammation caused by CCl4 by histopathology and enzyme-linked immunosorbent assay in mice. The expression of Akt/NF-κB inflammatory pathway was determined by real-time polymerase chain reaction (RT-PCR) and western blotting (WB). The results showed that COPs inhibited the expression of key proteins in the inflammatory pathway. In conclusion, the results of this study suggested that COPs could improve CCl4 -induced HF by improving liver injury, reducing the expression of inflammatory factors, and inhibiting the expression of inflammatory signaling pathways. PRACTICAL APPLICATIONS: The corns around the world are mainly used as animal feed, and the liver protective activity of corn oligopeptides (COPs) is rarely applied to the market. The development of COPs liver protective food can prevent the occurrence of liver-related diseases such as hepatic fibrosis to a certain extent. Developing COPs liver protecting food can improve the utilization value of corn. It is hoped that this study can provide experimental support for the application of COPs in liver protection food.

Caloric restriction inhibits renal artery ageing by reducing endothelin-1 expression.

BACKGROUND: The renal artery plays a central role in renal perfusion and is critical for proper renal function. Ageing is an independent risk factor for both impaired renal function and vascular disorders, and associated with an increase in the expression of the vasoconstrictor endothelin-1 (ET-1), and caloric restriction (CR) without malnutrition has been shown to be an effective inhibitor of renal dysfunction induced by ageing. The objective of this study was to determine whether CR-mediated alleviation of renal dysfunction is mediated by ET-1 expression. METHODS: The young (2 months, 2 M) and old (12 months, 12 M) Sprague-Dawley male rats were used and fed ad libitum. The 12-month-old rats were further divided into 12 M and 12 M-caloric restriction (CR) (30% calorie restriction). After 8 weeks, the renal tissues were showed by PAS staining, and age-related metabolic parameters and renal functions were detected in each group of rats. The inflammatory cytokines of interleukin (IL)-6, IL-1ß, tumor necrosis factor alpha (TNF-α), and transforming growth factor beta 1 (TGF-ß1) were analyzed using ELISA. The mRNA and protein expression in the renal artery were analysis by qRT-PCR and Immunoblot analysis. RESULTS: Ageing was associated with significant increases in 24 h urine protein content and serum triglyceride and cholesterol in 12 M rats, both of which were significantly inhibited in 12 M-CR. The mRNA expression and the secretion of IL-6, IL-1ß, TNF-α, and TGF-ß1 in the renal artery was significantly increased with ageing and inhibited by CR. CR also inhibited ageing-induced Edn1 (encoding ET-1) mRNA and protein expression in the renal artery. In addition, CR could regulate ET-1 expression by inhibiting the activation of NF-κB signaling and activation and induction in the expression of NF-E2-related factor 2 (Nrf2) and histone deacetylase and gene repressor sirtuin 1 (SIRT1), both of which play a central role in mitigating oxidative stress in young rats. CONCLUSIONS: Moderate CR can reverse the ageing related kidney dysfunction by reducing the ET-1 expression. CR might be used as an alternative to prevent the ageing induced renal artery dysfunction.

Effect of Shexiang Baoxin Pill () in Alleviating Early Hypertensive Renal Injury in Rats.

OBJECTIVE: To investigate the effect of Shexiang Baoxin Pill (, SBP) on early hypertensive renal injury in rats and to explore the possible mechanism. METHODS: Twelve-week-old spontaneous hypertensive rats (SHRs) with high-salt diet (dietary containing 8% NaCl) were randomized into the SBP group [40 mg/(kg·d)], losartan potassium group [20 mg/(kg·d)] and saline group by stratified random sampling method, 12 in each group. Blood pressure and urea albumin creatinine ratio were measured. After 10 weeks, the expression levels of serum creatinine (Scr), hypersensitive C-reactive protein (hs-CRP), interleukin (IL)-1 ß, IL-6, tumor necrosis factor α (TNF-α), and transforming growth factor ß (TGF-ß) in serum were assessed. Kidney pathology periodate-schiff staining was performed. Semi-quantitative count of macrophage infiltration was determined by immunochemistry of CD68 staining. Real-time quantitative polymerase chain reaction and Western blot were performed to examine the mRNA and protein expressions of Toll-like receptor 4 (TLR4), nuclear factor κB (NF-κB), monocyte chemokine peptide (MCP-1), inducible nitric oxide synthase (iNOS), and arginase-1 (Arg-1). RESULTS: SBP did not affect the mortality of SHR (P<0.05). SBP significantly reduced the level of elevated blood pressure of SHRs, but the effect was less significantly than that of losartan potassium. SBP decreased urine protein (P<0.01) and the expression levels of IL-1 ß, IL-6, TNF-α, and TGF-ß in serum. The 22-week-old SHRs showed mild proliferation of glomerular endothelial cells, glomerular ischemic lesions, inflammatory cell infiltration in renal tubular interstitium and arteriosclerosis. Both SBP and losartan potassium had alleviated renal pathological change, and significantly reduced the infiltration of macrophage (P<0.05, P<0.01). SBP and losartan potassium decreased the expressions of TLR4, NF-κB, MCP-1, iNOS, and Arg-1. CONCLUSION: SBP significantly modified the early hypertensive renal injury by reducing inflammation, and the effect was similar to losartan potassium.

[Isolation and Identification of Rat Kidney Stem Cells].

OBJECTIVE: To isolate and steadily culture kidney stem cells (KSCs) from rat renal papilla, and to identify the biological characteristics of KSCs. METHODS: KSCs were isolated from the tips of renal papilla in 4 weeks-old Sprague-Dawley rats. The morphology of KSCs was observed under inversion microscope, and the phenotye characteristics of kSCs were identified through flow cytometry and immunofluorescence. The abilities of KSCs in adipogenic and osteogenic differentiation were evaluated. The differences of gene expression between KSCs and rat renal tubular epithelial cells (RTECs)were compared using quantitative real time polymerase chain reaction (qRT-PCR). RESULTS: KSCs showed a spindle-shaped and arborization-like growth pattern. Immunofluorescence indicated that KSCs staining with alpha-sooth muscle actin (α-SMA), Vimentin, N-Cadherin, Nestin, CD133 marker, and without E-cadherin, cytokeratin-18 (CK-18), zona occludens protein-1 (ZO-1). The positive staining of CD29, CD90, CD73 were 99. 0%, 95. 8%, 99. 9% respectively, the positive staining of CD45 was 3. 4%. The positive stainings of stem cell marker CD133 and Nestin were 33. 2% and 70. 2% respectively, while the double staining rate was 31. 4%., KSCs showed positive staining by oil red 0 after adipogenic differentiation, and orange calcium deposition by alizarin red staining after osteogenic differentiation. qRT-PCR showed that the expressions of embryonic stem cell marker Nanog, Oct4/pou5f1,Sox2/sry-box-2 in KSCs were higher than those in RTECs (P< 0.01), and the expressions of mesenchymal marker c-SMA, Vimentin were also higher in KSCs (P<0. 01). Compared with RTECs, the expressions of mature epithelium marker E-Cadherin, CK18 in KSCs were lower (P< 0. 01). CONCLUSION: KSCs were isolated successfully and steadily cultured from the rat renal papilla, which were identified with featured biological characteristics.

Application of continuous renal replacement therapy for acute kidney injury in elderly patients.

This study aims to analyze the factors that affect the prognosis of continuous renal replacement therapy (CRRT) in elderly patients with acute kidney injury (AKI). Data obtained from 41 elderly patients with AKI who underwent CRRT in our department between January 2001 and December 2010 was retrospectively evaluated in this study. The enrolled patients were 80 to 100 years old, with a mortality of 60.98%. The mean Acute Physiology and Chronic Health Evaluation II (APACHE II) score was 27.8±5.6 points, and the mean risk coefficient was 0.80±0.10. The APACHE II score of the survival group was significantly higher than that of the death group. The comparisons of therapeutic dosages between <25 mL/(kg⋅h) and 25-50 mL/(kg⋅h), and between 25-50 mL/(kg⋅h) and >50 mL/(kg⋅h) all had no statistical significance. The prognosis of CRRT and the number of involved organs were related to the APACHE II score. Logistic regression analysis revealed that the number of involved organ, APACHE II score, mechanical ventilation, and hypoalbuminemia were the major risk coefficients that affected the prognosis of patients with bedside hemofiltration. The turnover of elderly CRRT patients was related to the number of involved organs, APACHE II score, mechanical ventilation, hypoalbuminemia, and other factors. The APACHE II score was the important reference index of CRRT starting time and could predict mortality risk.

[Risk factors and short-term prognosis of acute kidney injury in elderly patients].

OBJECTIVE: To explore the risk factors and short-term outcomes of acute kidney injury (AKI) in elderly patients. METHODS: A total of 232 elderly AKI patients at Chinese PLA General Hospital from June 2008 to December 2009 were enrolled. They were divided into two groups according to their outcomes at 28 days and at 29 days to 3 months after AKI respectively. Their clinical data were analyzed to explore the risk factors and their effects on the outcomes of AKI. RESULTS: There were 215 males and 17 females with an average age of (86.7 ± 5.3) years. Thirty-eight cases (16.4%) died within 28 days after AKI and 57 (24.6%) died within 3 months. Infection (43.1%) was the major cause of AKI. The other causes included hypovolemia (19.0%), use of nephrotoxic drugs (16.8%) and cardiovascular events (15.1%) respectively. Logistic regression analysis revealed that low body mass index (BMI), oliguria, mechanical ventilation, hypoalbuminemia and peak serum level of creatinine (Scr >246.5 µmol/L) were the prognostic factors of AKI in those patients dying within 28 days after AKI (P < 0.05). Low BMI, hypoalbuminemia and high blood level of urea nitrogen (BUN) were the prognostic factors of AKI in those patients dying within 29 days to 3 months after AKI (P < 0.05). CONCLUSION: Infection, hypovolemia, use of nephrotoxic drugs and cardiovascular events are common causes of AKI in elderly patients.Low BMI, oliguria, mechanical ventilation, hypoalbuminemia, high level of BUN and peak level of Scr ( > 246.5 µmol/L) are the prognostic factors of AKI in elderly patients.

[Effects of kidney stem cells on injure-recovery of human renal tubular epithelial cells].

OBJECTIVE: To investigate the protective effects and the mechanism of kidney stem cells (KSC) on the injured tubular epithelial cells. METHODS: KSC were isolated from rat renal papilla. The human kidney epithelial cells (HKC) injury model was induced with 0.1 µmol/L antimycin A for 30 minutes. The injured HKC were co-cultured with KSC or supernatant of cultured KSC. The apoptosis of HKC were detected by flow cytometry. The changes of ATP in the HKC and the level of malondialdehyde (MDA), superoxide dismutase (SOD) and lactate dehydrogenase (LDH) in the supernatant of cultured HKC were detected after being co-cultured with KSC. RESULTS: The study of the co-culture showed that KSC was less capable to migrate through the micropores of Transwell compared with bone marrow stem cells. However, after being co-cultured with the KSC conditional supernatant, ATP content of injured HKC, total SOD value in the supernatant of injured HKC were increased, and the MDA and LDH in the supernatant of injured HKC decreased. CONCLUSION: KSC have protected effects and participate in the repair of injured HKC.

[Stem cells and diabetic nephropathy].

Diabetic nephropathy (DN) is one of the most common microvascular complications of diabetic mellitus. Its treatment by now is mainly focused on the suppression of the renin-angiotonin system and the control of relative risk factors such as hypertension, hyperglycemia and so on, whose effectiveness is disappointed. Stem cells can be found in almost all organs and tissues, the main functional characteristics of stem cells are their capacity for self-renewal, the differentiation into several other cell types, and their immunomodulatory ability. In this article, we review the potential of stem cells as new therapeutic agents in the treatment of DN, discuss about how the diabetic environment affects these cells, and the therapeutic benefits that these stem cells offer for treating DN, suggesting that stem cells therapies might represent a new and promising strategy for the treatment of DN.

[Low testosterone levels are inversely correlated with carotid artery plaque formation in elderly women].

OBJECTIVE: To study the relationship between serum testosterone levels and the plaque formation of the carotid artery in a population-based cohort of independently living healthy women above 60 years of age. METHODS: Analysis of the healthy elders from a population-based cohort study in 9 communities of Beijing. Carotid intima-media thickness and atherosclerotic plaques were determined ultrasonographically. Serum testosterone levels were measured by immunoassay. The data were analyzed with ANOVA and logistic regression analysis. RESULTS: There was an inverse correlation between testosterone and plaque formation in old females (P < 0.01), while no association was found in males. Female with testosterone levels in the lowest quartile (< 0.49 nmol/L) had more risk of plaque formation (OR = 3.805, P < 0.01) after adjusted with age and other traditional factors of atherosclerosis. CONCLUSION: Testosterone concentrations are negatively associated with carotid artery atherosclerosis in old women in Beijing, experimental and prospective studies are needed to determine the possible therapeutic role of testosterone in atherosclerosis.

[Long-term follow-up of elderly patients with atherosclerotic renal artery stenosis treated by percutaneous transluminal angioplasty with stent implantation].

OBJECTIVE: To evaluate the long-term effects of percutaneous transluminal renal angioplasty with stent (PTRAS) on hypertension and renal function in elderly patients with atherosclerotic renal artery stenosis (ARAS). METHODS: The data of elderly ARAS patients as diagnosed by angiography (stenosis ≥ 70%) were retrospectively collected. PTRAS was performed in 65 patients. The average follow-up period was 30.9 months. RESULTS: There were significant decrease in BP (blood pressure, mm Hg, 1 mm Hg = 0.133 kPa) (before: 154 ± 24/ 79 ± 119 vs after: 132 ± 14/69 ± 8; P < 0.01) at Day 3 post-PTRAS and the decrease of BP continued until 36 months after PTRAS. The average category of antihypertensive medication also decreased from 2.3 ± 1.1 to 2.1 ± 1.0. The incidence of contrast-induced nephropathy (CIN) was 9.2%. Logistic regression analysis showed that the factors of pre-operative diabetes mellitus, GFR ≤ 30 ml×min(-1)·1.73 m(-2), systolic pressure ≥ 180 mm Hg and hydration therapy had a significant relationship with the incidence of CIN (P = 0.0072; OR = 13.51; P = 0.0002; OR = 519.27; P = 0.0134; OR = 13.16 and P = 0.0266; OR = 0.10; respectively). Renal function improved in 9.1%-15.8% of patients, stabilized in 67.3% - 55.3% and deteriorated in 23.4% - 28.9% of patients at Months 12 - 36 post-PTRAS. Logistic regression analysis showed that the diabetics had a higher risk of deteriorating renal function at month 12 post-PTRAS (P = 0.0277; OR = 6.32). The restenosis rate was 13.8%. CONCLUSION: PTRAS is beneficial in the control of blood pressure in elderly ARAS patients within 36 months after operation. The post-PTRAS improvement of renal function in elderly patients is limited.

[The effects of atorvastatin on aging kidney].

OBJECTIVE: To evaluate whether taking atorvastatin for long time has positive effects on age-related renal impairment. METHODS: 20-month-age normal female Wistar rats were divided into three groups (n = 9). First group were fed atorvastatin 10 mg/(kg x d). Second group were fed atorvastatin 1 mg/(kg x d). Third group were fed the same volume normal saline served as control. All the rats were sacrificed after four months. 3-month-age normal female Wistar rats (n = 9) also served as normal control. Kidney weight, serum creatinine (Scr) and blood-lipoids were measured. Paraffin sections of renal tissues were stained with PAS and Sirius red. Sclerosis index of glomerulus was calculated. RESULTS: Renal mass diminution was found in all the groups of aging rats. Scr was decreased in the group of aging rats with atorvastatin 1 mg/(kg x d). The level of blood-lipoids of aging rats was higher than that of young rats. The level of serum cholesterol and low-density lipoprotein (LDL) were decreased in first group (both P < 0.05) and only LDL decreased in second group (P < 0.05). Morphological changes of aging kidney were focal segmental glomerulosclerosis, widen of mesangial region, infiltration of inflammatory cells and sclerosis of arteriole. The treatment of atorvastatin improved the pathologic changes in the aging rats significantly, especially in the first group. CONCLUSION: Taking atorvastatin for long time can notably improve the pathological changes of aging kidney. All these effects may be induced by lowing of blood-lipoids, relieving the sclerosis of renal arteriole and reducing the infiltration of inflammatory cells.

Glomerular aging in females is a multi-stage reversible process mediated by phenotypic changes in progenitors.

The glomeruli of postmenopausal C57BL6 mice, and age-matched males, show progressive hypertrophy and glomerulosclerosis. We asked whether this was a multistage process, was due to alterations in glomerular progenitors, and was reversible in female mice. Using cross bone marrow transplants (BMT) between young and old females, we found that BMT delivered a phenotype that was donor age-specific. The fact that lesions in young recipients were more severe if the donors were in late rather than early menopause suggested that new progenitor phenotypes had appeared. Postmenopausal recipients of BMT from young donors had reduced glomerular hypertrophy and sclerosis, implying that the aging lesions in females were reversible and that progenitors, rather than the local environment, determined the glomerular profile. The altered phenotype included increased extracellular matrix synthesis and decreased matrix metalloproteinase-2 levels as well as cell hypertrophy. The mechanism of the cellular hypertrophy was due to uncoupling of hypertrophy from proliferation, resulting from elevated p27 levels. Thus, glomerular hypertrophy and sclerosis in aging females is a multistage process, is reversible, and may be determined by the phenotype of bone marrow-derived progenitor cells.

The glomerulosclerosis of aging in females: contribution of the proinflammatory mesangial cell phenotype to macrophage infiltration.

Age-associated renal changes may be an important cause of renal failure. We recently found that aged female B6 mice developed progressive glomerular lesions. This was associated with macrophage infiltration, a frequent finding in glomerulosclerosis. We used these mice as a model for studying the mechanisms of glomerular aging. We compared the gene expression profile of intact glomeruli from late postmenopausal (28-month-old) mice to that of intact glomeruli from premenopausal (5-month-old) mice. We found that inflammation-related genes, especially those expressed by activated macrophages, were up-regulated in the glomeruli of 28-month-old mice, a result correlating with the histological observation of glomerular macrophage infiltration. The mechanism for macrophage recruitment could have been stable phenotypic changes in mesangial cells because we found that mesangial cells isolated from 28-month-old mice expressed higher levels of RANTES and VCAM-1 than cells from 5-month-old mice. The elevated serum tumor necrosis factor (TNF)-alpha levels present in aged mice may contribute to increased RANTES and VCAM-1 expression in mesangial cells. Furthermore, cells from 28-month-old mice were more sensitive to TNF-alpha-induced RANTES and VCAM-1 up-regulation. The effect of TNF-alpha on RANTES expression was mediated by TNF receptor 1. Interestingly, mesangial cells isolated from 28-month-old mice had increased nuclear factor-kappaB transcriptional activity. Inhibition of nuclear factor-kappaB activity decreased baseline as well as TNF-alpha-induced RANTES and VCAM-1 expression in mesangial cells isolated from 28-month-old mice. Thus, phenotypic changes in mesangial cells may predispose them to inflammatory stimuli, such as TNF-alpha, which would contribute to glomerular macrophage infiltration and inflammatory lesions in aging.

Development of albuminuria and glomerular lesions in normoglycemic B6 recipients of db/db mice bone marrow: the role of mesangial cell progenitors.

The pathologic hallmarks of diabetic nephropathy are excess mesangial extracellular matrix (ECM) and mesangial cell proliferation. We previously showed that mesangial cell phenotypic changes play an important role in the pathogenesis of diabetic nephropathy. We concluded that phenotypic changes were present in bone marrow (BM)-derived mesangial cell progenitors, as transplantation of BM from db/db mice, a model of type 2 diabetic nephropathy, transferred the db genotype and a nephropathy phenotype to naive B6 mice recipients. The recipients did not develop diabetes; however, they did develop albuminuria and glomerular lesions mirroring those in the donors (i.e., glomerular hypertrophy, increased ECM, and increased cell number with cell proliferation). We found that matrix metalloproteinase 2 (MMP-2) facilitated invasion of the mesangial cells into ECM and proliferation in vitro. Thus, increased MMP-2 activity in db/db mesangial cell progenitors may partially explain increased mesangial cell repopulation and proliferation in B6 recipients of db/db BM. In summary, BM-derived mesangial cell progenitors may play a crucial role in the development and progression of ECM accumulation and mesangial cell proliferation in this model of diabetic nephropathy in type 2 diabetes.