scholarly journals The double edge of anti-CD40 siRNA therapy: It increases renal microcapillar density but favours the generation of an inflammatory milieu in the kidneys of ApoE−/− mice

2019 ◽  
Vol 16 (1) ◽  
Author(s):  
Miguel Hueso ◽  
Angela Casas ◽  
Adrian Mallén ◽  
Laura de Ramón ◽  
Nuria Bolaños ◽  
...  
Keyword(s):  
Renal Diseases ◽  
Physiological Basis ◽  
Sirna Treatment ◽  
Double Edge ◽  
Peritubular Capillaries ◽  
Sirna Silencing ◽  
Sirna Therapy ◽  
Inflammatory Milieu ◽  
Endothelial Dysfunctions ◽  
Target Effects

Abstract Background Chronic kidney disease (CKD) is associated with endothelial dysfunctions thus prompting links between microcirculation (MC), inflammation and major cardiovascular risk factors. Purpose of the study We have previously reported that siRNA-silencing of CD40 (siCD40) reduced atherosclerosis (ATH) progression. Here, we have deepened on the effects of the siCD40 treatment by evaluating retrospectively, in stored kidneys from the siCD40 treated ApoE−/− mice, the renal microcirculation (measured as the density of peritubular capillaries), macrophage infiltration and NF-κB activation. Methods Kidneys were isolated after 16 weeks of treatment with the anti-CD40 siRNA (siCD40), with a scrambled control siRNA (siSC) or with PBS (Veh. group). Renal endothelium, infiltrating macrophages and activated NF-κB in endothelium were identified by immunohistochemistry, while the density of stained peritubular capillaries was quantified by image analysis. Results ATH was associated with a reduction in renal MC, an effect reversed by the anti-CD40 siRNA treatment (3.8 ± 2.7% in siCD40; vs. 1.8 ± 0.1% in siSC; or 1.9 ± 1.6% in Veh.; p < 0.0001). Furthermore, siCD40 treatment reduced the number of infiltrating macrophages compared to the SC group (14.1 ± 5.9 cells/field in siCD40; vs. 37.1 ± 17.8 cells/field in siSC; and 1.3 ± 1.7 cells/field in Veh.; p = 0.001). NF-κB activation also peaked in the siSC group, showing lower levels in the siCD40 and Veh. groups (63 ± 60 positive cells/section in siCD40; vs. 152 ± 44 positive cells/section in siSC; or 26 ± 29 positive cells/section in veh.; p = 0.014). Lastly, serum creatinine was also increased in the siCD40 (3.4 ± 3.3 mg/dL) and siSC (4.6 ± 3.0 mg/dL) groups when compared with Veh. (1.1 ± 0.9 mg/dL, p = 0.1). Conclusions Anti-CD40 siRNA therapy significantly increased the density of peritubular capillaries and decreased renal inflammation in the ATH model. These data provide a physiological basis for the development of renal diseases in patients with ATH. Furthermore, our results also highligth renal off-target effects of the siRNA treatment which are discussed. Graphical abstract

Single and Combinational siRNA Therapy of Cancer Cells: Probing Changes in Targeted and Nontargeted Mediators after siRNA Treatment

2016 ◽  
Vol 13 (12) ◽  
pp. 4116-4128 ◽  
Author(s):  
Hamidreza Montazeri Aliabadi ◽  
Parvin Mahdipoor ◽  
Marco Bisoffi ◽  
Judith C. Hugh ◽  
Hasan Uludağ
Keyword(s):  
Cancer Cells ◽  
Sirna Treatment ◽  
Sirna Therapy

Abstract 021: Haplotype-Dependent Differential Regulation Of The Human AT1R Gene Is Exacerbated By Age: Effects On Tissue Inflammatory And Redox Milieu

Hypertension ◽  
2014 ◽  
Vol 64 (suppl_1) ◽  
Author(s):  
Anita Rana ◽  
Sudhir Jain ◽  
Nitin Puri ◽  
Brahmaraju Mopidevi ◽  
Meenakshi Kaw ◽  
...  
Keyword(s):  
Blood Pressure ◽  
Transcriptional Regulation ◽  
Gene Expression Regulation ◽  
Artificial Chromosome ◽  
Organ Damage ◽  
Antioxidant Defenses ◽  
Renal Diseases ◽  
R Gene ◽  
Nucleotide Polymorphisms ◽  
Inflammatory Milieu

Age-associated inflammation and redox imbalance underlie etiopathogenesis of cardiovascular-renal diseases, including hypertension and end organ damage. Angiotensin II (Ang II), via activation of the AT1R, contributes to the development and progression of these pathophysiologies. We have identified two haplotype blocks of single nucleotide polymorphisms (SNPs) in the hAT 1 R gene: haplotype II (Hap II: -810A, -713G, -214C, -153G) and I (Hap I: -810T, -713T, -214A, -153A). In clinical studies, Hap I is linked to human hypertension. This study examines haplotype-dependent and age-associated transcriptional regulation of the hAT1R gene. In this regard, we have engineered transgenic (TG) mice with either haplotype of the hAT1R gene using a 166-kb bacterial artificial chromosome. ChIP assay shows increased RNA-Pol II binding (~1.6 fold higher) to the chromatin extracts from renal tissues of adult (4-6 months) male Hap I-TG mice with increased hAT1R expression (~6 fold higher). This was accompanied by higher baseline blood pressure in Hap I-TG mice (Hap I- 129±3 vs. Hap II- 116±4, p<0.05). Next, we examined the effects of age on this haplotype-dependent regulation of the hAT1R. Aged (>18 months), male mice were used for this part of the study. hAT1R expression increases with age in both haplotypes; however, this increase is significantly higher in Hap I-TG mice (3.17±0.5 to 5.5±0.75 fold) as opposed to mice with Hap II (1.1±0.2 to 1.8±0.1 fold). Age-associated change (Δ) in inflammatory and redox markers was significantly (p<0.05) greater in TG mice with Hap I including, IL1 (4.6±0.8 vs. 2.1±0.49 fold), IL6 (4.0±0.69 vs. 2.1±0.2 fold) and NOX1 (8.3±0.4 vs. 2.5±0.6 fold). This is accompanied by age-associated reduction in levels of antioxidant defenses (SOD1: 0.97±0.0 vs. 1.4±0.1 fold; HO1: 0.77±0.1 vs. 1.3±0.2 fold) and pro-survival genes including, NAMPTS (2.1 folds lower in Hap I vs. Hap II) and SIRT1 (1.8 fold lower in Hap I vs. Hap II). Thus, haplotype-dependent transcriptional regulation of the hAT 1 R gene causes increased hAT1R expression and blood pressure, in Hap I TG mice. Importantly, aging exacerbates this differential gene-expression regulation, further increasing hAT1R and promoting a prooxidant/inflammatory milieu in mice with Hap I.

scholarly journals Prostacyclin Synthase: Upregulation during Renal Development and in Glomerular Disease as well as Its Constitutive Expression in Cultured Human Mesangial Cells

2015 ◽  
Vol 2015 ◽  
pp. 1-9 ◽  
Author(s):  
Thomas Klein ◽  
Günther Klaus ◽  
Martin Kömhoff
Keyword(s):  
Mesangial Cells ◽  
Constitutive Expression ◽  
Glomerular Disease ◽  
Renal Diseases ◽  
Prostaglandin E ◽  
Human Mesangial Cells ◽  
Immunoreactive Protein ◽  
Peritubular Capillaries ◽  
Prostacyclin Synthase

Prostacyclin (PGI2) plays a critical role in nephrogenesis and renal physiology. However, our understanding of how prostacyclin release in the kidney is regulated remains poorly defined. We studied expression of prostacyclin synthase (PGIS) in developing and adult human kidneys, and also in selected pediatric renal diseases. We also examined PGI2formation in human mesangial cellsin vitro. We observed abundant expression of PGIS in the nephrogenic cortex in humans andin situhybridization revealed an identical pattern in mice. In the normal adult kidney, PGIS-immunoreactive protein and mRNA appear to localize to mesangial fields and endothelial and smooth muscle cells of arteries and peritubular capillaries. In kidney biopsies taken from pediatric patients, enhanced expression of PGIS-immunoreactive protein was noted mainly in endothelial cells of patients with IgA-nephropathy. Cultured human mesangial cells produce primarily PGI2and prostaglandin E2, followed by prostaglandin F2αCytokine stimulation increased PGI2formation 24-fold. Under these conditions expression of PGIS mRNA and protein remained unaltered whereas mRNA for cyclooxygenase-2 was markedly induced. In contrast to its constitutive expressionin vitro, renal expression of prostacyclin-synthase appears to be regulated both during development and in glomerular disease. Further research is needed to identify the factors involved in regulation of PGIS-expression.

scholarly journals Enhancing Chemotherapy by RNA Interference

2020 ◽  
Vol 1 (2) ◽  
pp. 64-81
Author(s):  
Shuwen Cao ◽  
Chunhao Lin ◽  
Shunung Liang ◽  
Chee Hwee Tan ◽  
Phei Er Saw ◽  
...  
Keyword(s):  
Anticancer Drugs ◽  
Sequential Therapy ◽  
Sirna Treatment ◽  
Rnai Technology ◽  
New Strategy ◽  
Multiple Challenges ◽  
Sirna Therapy ◽  
Interfering Rna ◽  
Target Side ◽  
Specific Mrna

Abstract Small interfering RNA (siRNA) has shown tremendous potential for treating human diseases in the past decades. siRNA can selectively silence a pathological pathway through the targeting and degradation of a specific mRNA, significantly reducing the off-target side effects of anticancer drugs. However, the poor pharmacokinetics of RNA significantly restricted the clinical use of RNAi technology. In this review, we examine in-depth the siRNA therapeutics currently in preclinical and clinical trials, multiple challenges faced in siRNA therapy, feasibility of siRNA treatment with anticancer drugs in combined with siRNA in nanoparticles or modified to be parental drugs, sequential therapy of siRNA treatment prior to drug treatment with siRNA and drugs loaded in nanoparticles. We focused on the combinatorial activation of apoptosis by different pathways, namely Bcl-2, survivin, and Pgp protein. Taken together, this review would serve to establish the pathway of effective and efficient combination therapy of siRNA and drugs as a new strategy.

scholarly journals Effects of survivin interference RNA on non-small cell lung carcinoma

2009 ◽  
Vol 32 (6) ◽  
pp. 225 ◽  
Author(s):  
Guan-Feng Liu ◽  
Qi-Gang Zhao ◽  
Lei Si ◽  
Yin-Guang Cao ◽  
Guang-Yao Li ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Tumour Growth ◽  
Human Lung ◽  
Small Cell ◽  
Human Lung Cancer ◽  
Small Cell Lung ◽  
Sirna Treatment ◽  
Sirna Therapy ◽  
Interference Rna

Objectives: The primary purpose of this study was to investigate the in vitro and in vivo effect of survivin interference RNA (siRNA) on non-small cell lung cancer. Methods: Lentivirus was used as a vector to transfer siRNA into human lung cancer A549 cells. The proliferation of the cancer cells was assessed by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. The lentivirus-mediated siRNA was also injected into the transplanted A549 tumor tissues in mice. Tumour growth was assessed after 11 injections over a period of 21 days. Results: Compared with the placebo and the blank lentiviral vector groups, the siRNA treatment group had reduced cell growth rate following 4 days of the treatment (P < 0.01). The average size of the transplanted A549 tumours in the siRNA treatment group (0.75±0.16 cm3, n=8) was smaller than in the placebo (2.09±0.22 cm3, n=6) or the blank lentivrial vector groups (1.89±0.18 cm3, n=6) (P < 0.01). The tumour growth inhibition rate in the siRNA groups was 46.1%. Conclusion: Lentivirus-mediated siRNA therapy inhibits the growth of human lung cancer cells in vitro. The siRNA therapy also suppresses the growth of the transplanted lung cancer in mice.

Physical Properties of Isolated Perfused Renal Tubular Basement Membranes

1971 ◽  
Vol 29 ◽  
pp. 390-391
Author(s):  
Jared Grantham ◽  
Larry Welling
Keyword(s):  
Basement Membrane ◽  
Basement Membranes ◽  
Transport Mechanisms ◽  
Permeability Characteristics ◽  
Peritubular Capillaries ◽  
Strategic Location ◽  
Tubular Lumen ◽  
Glomerular Filtrate ◽  
Urine Formation ◽  
Renal Tubular

In the course of urine formation in mammalian kidneys over 90% of the glomerular filtrate moves from the tubular lumen into the peritubular capillaries by both active and passive transport mechanisms. In all of the morphologically distinct segments of the renal tubule, e.g. proximal tubule, loop of Henle and distal nephron, the tubular absorbate passes through a basement membrane which rests against the basilar surface of the epithelial cells. The basement membrane is in a strategic location to affect the geometry of the tubules and to influence the movement of tubular absorbate into the renal interstitium. In the present studies we have determined directly some of the mechanical and permeability characteristics of tubular basement membranes.

A rapid silver-impregnation technique on ultra-thin sections for Electron Microscopy

1993 ◽  
Vol 51 ◽  
pp. 242-243
Author(s):  
K. Chien ◽  
R.C. Heusser ◽  
M.L. Jones ◽  
R.L. Van de Velde
Keyword(s):  
Electron Microscopy ◽  
Silver Nitrate ◽  
Silver Impregnation ◽  
Sodium Borate ◽  
Renal Diseases ◽  
Distilled Water ◽  
Thin Sections ◽  
Impregnation Technique ◽  
Centrifuge Tube ◽  
Simplified Procedure

Silver impregnation techniques have been used for the demonstration of the complex carbohydrates in electron microscopy. However, the silver stains were believed to be technically sensitive and time consumming to perform. Currently, due to the need to more specifically evaluate immune complex for localization in certain renal diseases, a simplified procedure in conjunction with the use of the microwave has been developed and applied to renal and other biopsies. The procedure is as follows:Preparation of silver methenamine solution:1. 15ml graduated, clear polystyrene centrifuge tube (Falcon, No. 2099) was rinsed once with distilled water.2. 3% hexamethylene tetramine (methenamine) was added into the centrifuge tube to the 6ml mark.3. 3% silver nitrate was added slowly to the methenamine to the 7ml mark while agitating. (Solution will instantly turn milky in color and then clear rapidly by mixing. No precipitate should be formed).4. 2% sodium borate was added to the solution to the 8ml mark, mixed and centrifuged before use.

How to Psychologically Minimize Scratching Impulses

2014 ◽  
Vol 222 (3) ◽  
pp. 140-147 ◽  
Author(s):  
Ariane Sölle ◽  
Theresa Bartholomäus ◽  
Margitta Worm ◽  
Regine Klinger
Keyword(s):  
Cognitive Processes ◽  
Placebo Effect ◽  
Psychological Factors ◽  
Pharmacological Treatments ◽  
Physiological Basis ◽  
Psychological Mechanisms ◽  
Physical Complaints ◽  
Factors Influencing ◽  
The Impact ◽  
Placebo Model

Research in recent years, especially in the analgesic field, has intensively studied the placebo effect and its mechanisms. It has been shown that physical complaints can be efficiently reduced via learning and cognitive processes (conditioning and expectancies). However, despite evidence demonstrating a large variety of physiological similarities between pain and itch, the possible transfer of the analgesic placebo model to itch has not yet been widely discussed in research. This review therefore aims at highlighting potential transfers of placebo mechanisms to itch processes by demonstrating the therapeutic issues in pharmacological treatments for pruritus on a physiological basis and by discussing the impact of psychological mechanisms and psychological factors influencing itch sensations.

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