Adriamycin-induced Nephropathy In Rats Research Articles

Epimedium sagittatum Maxim ameliorates adriamycin-induced nephropathy by restraining inflammation and apoptosis via the PI3K/AKT signaling pathway.

Modern pharmacological studies show that Epimedium sagittatum Maxim (EPI) has antioxidant, antiapoptotic, anti-inflammatory effects. However, the effects of EPI on adriamycin-induced nephropathy are unclear. The main purpose of this study is to investigate the effects of EPI on adriamycin-induced nephropathy in rats. The chemical composition of EPI was detected by high performance liquid chromatography. Network pharmacology was used to collect the effects of EPI on adriamycin nephropathy; renal histological changes, podocyte injury, inflammatory factors, oxidative stress levels, apoptosis levels, and the PI3K/AKT signaling pathway were examined. Moreover, analyze the effects of icariin (the representative component of EPI) on adriamycin-induced apoptosis and PI3K/AKT signaling pathway of NRK-52e cells. Network pharmacological results suggested that EPI may ameliorate adriamycin-induced nephropathy by inhibiting inflammatory response and regulating the PI3K/AKT signaling pathway. The experimental results showed that EPI could improve pathological injury, renal function, podocyte injury, and inhibit inflammation, oxidative stress, apoptosis in adriamycin-induced nephropathy rats through the PI3K/AKT signaling pathway. Furthermore, icariin inhibited adriamycin-induced mitochondrial apoptosis in NRK-52e cells. This study suggested that EPI ameliorates adriamycin-induced nephropathy by reducing inflammation and apoptosis through the PI3K/AKT signaling pathway, icariin may be the pharmacodynamic substance basis for this effect.

Tetrandrine alleviates podocyte injury via calcium-dependent calpain-1 signaling blockade

BackgroundPodocytes have become a crucial target for interventions in proteinuric kidney diseases. Many studies have reported that overexpression of transient receptor potential cation channel protein 6 (TRPC6) in podocyte injury upregulates intracellular Ca2+ influx and stimulates Ca2+-dependent protease calpain-1 signaling. The traditional Chinese drug, tetrandrine, a nonselective Ca2+ channel blocker, has long been used to treat chronic kidney disease. This research aimed to explore the possible mechanisms underlying the anti-proteinuric properties of tetrandrine.MethodsWe investigated the involvement of tetrandrine in Ca2+ dependent calpain-1 signaling in mouse podocytes and adriamycin-induced nephropathy rats. Cyclosporine A (CsA) and U73122 were used as positive controls. Cell viability, cytotoxicity, Ca2+ concentration, calpain activity, and mRNA and protein expression levels of calpain-1 signaling pathways were examined. The clinical and pathological changes were measured.ResultsTetrandrine decreased intracellular Ca2+ influx in cultured TRPC6-overexpressing podocytes. In both in vitro and in vivo studies, the administration of tetrandrine downregulated calpain activity and the expression of calpain-1 and restored the expression of downstream Talin-1 and nephrin. Compared to CsA, tetrandrine treatment exhibited superior inhibitory effects on calpain activity and calpain-1 expression.ConclusionsTetrandrine has therapeutic potential in podocyte damage by blocking Ca2+-dependent activation of the calpain-1 signaling pathway. Tetrandrine reduced proteinuria, improved renal function, and alleviate renal pathological damage.

Ultra-Performance Liquid Chromatography-Q-Exactive Orbitrap-Mass Spectrometry Analysis for Metabolic Communication between Heart and Kidney in Adriamycin-Induced Nephropathy Rats

Background/Aims: Although the adriamycin-induced nephropathy model is frequently employed in the study of nephrotic syndrome and focal segmental glomerulosclerosis, the accompanying myocardial damage has always been a cause for concern. Therefore, there is a great need to study cardiorenal communication in this model. Methods: An adriamycin-induced nephropathy model was established via tail vein injection. The levels of the biochemical indicators serum albumin, serum globulin, serum total protein, serum cholesterol, serum creatinine (SCr), urinary protein, and urinary creatinine (UCr) were measured, and histopathological changes in the heart and kidneys were assessed using hematoxylin-eosin staining. Metabolomic changes in the heart, blood, and kidneys were analyzed using the metabolomics method based on ultra-performance liquid chromatography Q-Exactive Orbitrap mass spectrometry. Results: Compared with the control group, the model group showed significant decreases in serum protein and total protein levels, albumin/globulin ratio, and creatinine clearance rate as well as significant increases in serum cholesterol, SCr, urinary protein, and UCr levels. Significant pathological changes were observed in the renal pathology sections in the model group, including diffusely merged glomerular epithelial cells, inflammatory infiltration, and vacuolated glomerular cells. Additionally, thickened myocardial fibers, swollen nuclei, inflammatory infiltration, and partial myocardial necrosis could be seen in the cardiac pathology sections in the model group. Based on multivariate statistical analysis, a total of 20 differential metabolites associated with 15 metabolic pathways were identified in the heart, 7 differential metabolites with 7 metabolic pathways were identified in the blood, and 16 differential metabolites with 21 metabolic pathways were identified in the kidney. Moreover, 6 common metabolic pathways shared by the heart and kidney were identified: arginine and proline metabolism; arginine biosynthesis; glutathione metabolism; alanine, aspartate, and glutamate metabolism; beta-alanine metabolism; and histidine metabolism. Among these metabolic pathways, alanine, aspartate, and glutamate metabolism was shared by the heart, blood, and kidney. Succinic acid was found to be the key regulatory metabolite in cardiorenal metabolic communication. Conclusion: Six metabolic pathways were found to be involved in cardiorenal metabolic communication in an adriamycin-induced nephropathy model, in which alanine, aspartate, and glutamate metabolism may be the metabolic link between the heart and kidney in the development and maintenance of oxidative stress and inflammation. Succinic acid may serve as a key regulatory metabolic switch or marker of cardiac and renal co-injury, as shown in an adriamycin-induced nephropathy model.

Ultrasound combined with microbubbles enhances the renoprotective effects of methylprednisolone in rats with adriamycin-induced nephropathy

The purpose of this study was to investigate the effect of ultrasound combined with microbbules (SonoVueTM) on the potency of methylprednisolone in attenuating the renal injury induced by adriamycin in rats. Animal model was established by two intravenous injections of 4 mg/kg adriamycin with a 2-week interval in rats. One week later, the adriamycin injected rats were randomly divided into 7 groups, receiving various treatments daily for 2 weeks. Two doses of methylprednisolone (20 or 40 mg/kg) were administrated alone or 20 mg/kg methylprednisolone and 100 µL SonoVueTM microbbules (1–5 × 108 bubbles/mL; mean diameter of bubbles: 2.5 µm) was co-administrated by intravenous injections from the tail vein. The ultrasound was applied at a frequency of 0.8 MHz and a spatial average temporal average intensity of 2.79 W/cm2 for 5 min at a 50% duty cycle (1 s on 1 s off) on the back skin of the anatomic position of the kidney in rats of two groups combined with ultrasound. Renal injury were analyzed using immunohistochemical staining, real-time PCR, light and transmission electron microcopies. The kidney function related biochemical indexes were measured by automatic biochemistry analyzer. The results showed that adriamycin induced a typical renal injury and 40 mg/kg methylprednisolone injection significantly ameliorated the abnormality of key parameters such as proteinuria, renal mRNA and protein expression levels of nephrin, collagens III and IV as well as podocyte impairment, glomerulosclerosis and tubulointerstitial injury indexes. However, a sub-dose of methylprednisolone at 20 mg/kg was ineffective when administered intravenously, but its potency at this dosage was enhanced by co-administration with 100 µL SonoVueTM microbubbles plus ultrasound irradiation. In conclusion, ultrasound combined with microbubbles can significantly increase local renal drug delivery leading to enhanced therapeutic effect of low dose methylprednisolone in ameliorating adriamycin-induced nephropathy in rats.

CTLA4-Ig Abatacept Ameliorates Proteinuria by Regulating Circulating Treg/IL-17 in Adriamycin-Induced Nephropathy Rats

Objective This study is aimed at investigating the efficacy of CTLA4-Ig abatacept in normalizing proteinuria and its possible mechanism in adriamycin-induced nephropathy (AIN) rats. Methods A total of 32 healthy male Sprague-Dawley rats were randomly divided into a normal group, an AIN group, an abatacept group, and a prednisone group. Adriamycin (6.5 mg/kg) was injected once via the tail vein of rats to induce nephrotic syndrome. After adriamycin treatment, the abatacept group rats were given abatacept (0.5 mg/kg) once by intraperitoneal injection on day 14. In addition, the prednisone group rats were given prednisone (12.5 mg/kg) daily consecutively by gavage from day 14 to day 21. Blood, urine, and kidney tissue specimens were collected when sacrificed on day 21. The 24-hour urinary protein, serum albumin, cholesterol, creatinine, and urea nitrogen were then detected. An enzyme-linked immunosorbent assay was used to determine the level of urine CD80 and serum IL-17. Flow cytometry was used to investigate the prevalence of circulating Treg. Hematoxylin-eosin staining and electron microscopy were used for a renal histological study. Immunofluorescence staining was performed to confirm the CD80 expression of renal tissue. Results The 24-hour urinary protein of the abatacept group was significantly lower than that of the prednisone group and the AIN group. The level of urine CD80 of the abatacept group was significantly lower than that of the AIN group. Compared with the AIN group and the prednisone group, the circulating Treg prevalence of the abatacept group was significantly higher, while the level of serum IL-17 was lower. A negative kidney staining of CD80 expression was demonstrated in each group in this study. The 24-hour urinary protein had a negative correlation with the circulating Treg prevalence and Treg/IL-17 and a positive correlation with the urine CD80 and serum IL-17. Urinary CD80 had a positive correlation with serum IL-17 and no correlation with the circulating Treg prevalence. Conclusions CTLA4-Ig abatacept can reduce proteinuria of adriamycin-induced nephropathy rats, possibly at least partially as a result of regulating circulating Treg/IL-17. CTLA4-Ig abatacept could be a promising regimen for idiopathic nephrotic syndrome.

Effects of prednisolone on adriamycin-induced nephropathy in rats

Effects of prednisolone on adriamycin-induced nephropathy in rats

Forsythiaside A alleviates renal damage in adriamycin-induced nephropathy.

Nephrotic syndrome (NS) is a common urinary system disease that carries a poor progrnosis due to nonresponsivess to current treatments. The purpose of this study was to investigate the therapeutic effects of Forsythiaside (FA) in a rat model of adriamycin-induced nephropathy (AN). Compared with control group, FA treatment significantly reduced proteinuria, serum creatinine and urea nitrogen levels and reduced the number of apoptotic cells in the kidneys. FA alleviated Adr induced renal injury, and increased the sactivity of superoxide dismutase (SOD), while decreasing malondialdehyde (MDA) and lactate dehydrogenase (LDH) in the serum. In addition, FA, in a dose-dependent manner, decreased the levels of NF-kappaBp65/MIP-2 in the kidenys, reduced the serum level of pro-inflammatory cytokines (IL-6, IL-1 β and TNF-alpha), and increased the survival rate of Adr treated rats. Taken together, the results show that FA alleviates renal dysfunction in adriamycin-induced nephropathy rats.

Evaluation of Injury Degree of Adriamycin-Induced Nephropathy in Rats Based on Serum Metabolomics Combined with Proline Marker.

Nephrotic syndrome (NS) is one of the leading causes of end-stage renal failure. Unfortunately, reliable surrogate markers for early diagnosing and monitoring the entire progression of NS are as yet absent. A method using UPLC-Q exactive HR-MS was established for the serum metabolomic study of adriamycin-induced nephropathy in rats. Two rat nephropathy models induced by adriamycin were adopted to reflect different degrees of renal damage of early and advanced stages. Then two MPC5 cell models were used to verify the role of proline in the progression of kidney injury. The results showed that seven metabolites such as 14S-HDHA, DPA, and DHA were associated with early renal injury, while 12 metabolites such as tryptophan, linoleyl carnitine, and LysoPC (18:3) reflected the advanced renal disease. At the same time, metabolites including LPE (22:6), LysoPC (22:5), and proline that changed during the whole process of NS were defined as progressive markers. Pathway analysis results showed that fatty acid metabolism, glycerophospholipid metabolism, and amino acids metabolism participated in the occurrence and development of NS. In addition, the change trend of intracellular proline content was consistent with that in serum, and the results were further supported by the detection of the crucial gene PYCRL. This study provides an important basis for searching for diagnostic markers of NS and also provides a methodological reference for early diagnosing and monitoring the pathogenesis of other progressive diseases.

Effects of sequential application of immunosuppressive agents according to the cell cycle in adriamycin-induced nephropathy rats.

The aim of this study was to observe the therapeutic effects of three sequential drug-based treatments according to the cell cycle in rats with adriamycin-induced nephropathy. A rat model of adriamycin-induced nephropathy was prepared by two injections, and three experimental groups were set up: control group (n=8); adriamycin-induced nephropathy rat group (n=8); and Meprednisone (MP), Ciclosporin (CsA), and mycophenolate (MMF) treatment group (n=8). Twenty-four-hour urine protein was quantified and serum total protein (TP), albumin (ALB), cholesterol (Chol), triglyceride (TG), urea nitrogen (BUN), and serum creatinine (Scr) were measured by an automatic biochemical analyzer. Pathological changes in renal tissues were observed by light microscopy. Serum matrix metalloproteinase-2 (MMP-2), MMP-9, and transforming growth factor-β1 (TGF-β1) were evaluated by double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Connective tissue growth factor (CTGF) expression was measured by Western blotting. Expression of nephrin and podocin in podocytes was compared by immunohistochemistry and quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Compared with the control group, 24-h urine protein in nephropathy group was significantly reduced at 2, 4, 8, and 12 weeks (p<0.01). Twenty-four-hour urine protein in the three treatment groups was significantly decreased compared with nephropathy group at 8 and 12 weeks (p<0.05). There were no significant differences among treatment groups (p>0.05), but their levels were higher than those in control group. TP and ALB levels in nephropathy group were decreased compared with control group (p<0.01) and increased compared with treatment groups (p<0.05), while TG and Chol levels in nephropathy group were increased compared with control group (p<0.01) and decreased compared with treatment group (p<0.05). There were no significant differences in biochemical parameters among the treatment groups. TGF-β1 levels were decreased, MMP-2 and MMP-9 levels were increased, and CTGF expression was reduced in the three therapeutic groups. Among the treatment groups, the combination of MP, CsA, and Rapa significantly inhibited fibrosis. The protein and mRNA levels of nephrin and podocin were significantly decreased in nephropathy group and their expression and distribution were partially restored in treatment groups. The present findings suggest that the sequential therapeutic treatments based on the cell cycle significantly improved the pathological changes in adriamycin-induced nephropathy rats. The sequential treatments significantly reduced urine protein levels, increased TP, ALB, MMP-2, and MMP-9 levels, decreased TG, Chol, and TGF-β1 levels, restored expression of nephrin and podocin in renal tissues, and significantly improved renal fibrosis.

Effects of Pravastatin in Adriamycin-Induced Nephropathy in Rats.

The aim of this study is to evaluate the effects of pravastatin on Adriamycin (ADR)-induced nephropathy and the mechanisms involved. Forty rats were divided into the following 4 groups: control, ADR (15 mg.kg-1, IP), ADR plus pravastatin (20 mg.kg-1 which was started 5 days prior to ADR injection), and ADR plus pravastatin (20 mg.kg-1 which was started 5 days after ADR injection). On day 20 after ADR injection, the animals were sacrificed. The results showed that administration of pravastatin decreased the levels of 24-h urinary protein (24-h UP), blood urea nitrogen (BUN), and creatinine (p < 0.05) which had increased after the injection of ADR; in addition, pravastatin reversed structural changes seen in ADR group. Furthermore, pravastatin elevated mRNA and protein expression of nephrin (p < 0.05) which had been reduced in ADR group. We conclude that pravastatin protects and treats renal injury induced by ADR.

Rituximab protects podocytes and exerts anti-proteinuric effects in rat adriamycin-induced nephropathy independent of B-lymphocytes.

This study was aimed at examining the effects of treatment with rituximab, a chimeric monoclonal antibody against the protein CD20, in a B-lymphocyte independent adriamycin-induced rat model of nephrotic syndrome. Rituximab is an emerging rescue therapy used in patients with complicated nephrotic syndrome and, therefore, we sought to elucidate the apparent B-lymphocyte independent mechanism underlying its anti-proteinuric effect. Adriamycin-induced nephropathy was established in Wistar rats by intravenously injecting 10 mg/kg of adriamycin, which were then treated with rituximab or purified human IgG weekly and euthanized on day 28. Proteinuria, glomerular expression of sphingomyelin phosphodiesterase acid-like 3b protein, and podocyte-related proteins were examined using immunofluorescence staining and a reverse transcription-polymerase chain reaction. Rituximab treatment of rats with adriamycin-induced nephropathy significantly reduced urinary protein excretion 14 to 28 days after induction of disease, compared with those treated with purified normal human IgG. Furthermore, rituximab treatment also prevented the reduction of glomerular nephrin and podocin expression on day 28. Double-immunofluorescence staining revealed that after in vivo administration, rituximab was bound to the glomeruli, which also expressed synaptopodin or sphingomyelin phosphodiesterase, acid-like 3b. Moreover, sphingomyelin phosphodiesterase, acid-like 3b expression was significantly decreased on day 28 of adriamycin-induced nephropathy, which was also prevented by rituximab. This study demonstrated that rituximab directly affected glomerular podocytes and ameliorated proteinuria in adriamycin-induced nephropathy in rats. Furthermore, protection of podocyte function by rituximab may be mediated by direct modulation of a sphingomyelin phosphodiesterase, acid-like 3b-dependent mechanism.

Matrine ameliorates adriamycin-induced nephropathy in rats by enhancing renal function and modulating Th17/Treg balance

Matrine (MAT) is an active alkaloid extracted from Radix Sophora flavescens. The present study was to investigate whether MAT could effectively treat Adriamycin-induced nephropathy (AIN). AIN was induced in rats using a single injection of Adriamycin (ADR). Renal interleukin-6 (IL-6), IL-10, IL-17 and transforming growth factor-β (TGF-β) levels, and the expression of forkhead box protein 3 (Foxp3) and retinoid-related orphan nuclear receptor γt (Rorγt) was measured. AIN rats developed severe albuminuria, hypoalbuminaemia, hyperlipidaemia and podocyte injury. Daily administration of MAT (100mg/kg or 200mg/kg) significantly prevented ADR-induced podocyte injury, decreased AIN symptoms and improved renal pathology manifestations. Of note, treatment with MAT (100mg/kg) plus prednisone (Pre, 5mg/kg) had equivalent efficacy to that of Pre alone (10mg/kg). Additional findings showed that ADR triggered a disordered cytokine network and abnormal expression of Foxp3 and Rorγt in rats, as reflected by increased levels of IL-6, IL-10, TGF-β, Rorγt and decreased levels of IL-10 and Foxp3. Interestingly, MAT weakened the disordered cytokine network and normalized the expression of Foxp3 and Rorγt. In addition, a significant negative correlation was observed between the values of Foxp3/Rorγt and renal pathology scores. Finally, MAT normalized regulatory T cells (Treg)/ T-helper17 cells (Th17) ratio in peripheral blood mononuclear cells of AIN rats. These data indicate MAT prevents AIN through the modification of disordered plasma lipids and recovery of renal function, and this bioactivity is at least partly attributed to the suppression of renal inflammation and the regulation of the Treg/Th17 imbalance.

Effects of Shenkangling intervention on the MAPK pathway in rats with doxorubicin-induced nephropathy.

Shenkangling plays a role of Yishenhuoxue effect for the treatment of children with nephrotic syndrome. The aim of this study was to investigate the effects of Shenkangling intervention on the mitogen-activated protein kinase (MAPK) pathway in rats with Adriamycin-induced nephropathy (AN) and its underlying mechanism of action. Nephrosis was induced in healthy Sprague-Dawley rats by doxorubicin and the rats were untreated or treated with prednisone, simvastatin, Shenkangling, or a combination thereof. Using real-time PCR, the mRNA expression levels of Chemokine (C-X-C motif) ligand 16 (CXCL16), A Disintegrin and metalloproteinase domain-containing protein 10 (ADAM10), and ADAM17 in the renal tissues of these rats were found to be decreased by the various treatments compared to those in the untreated doxorubicin-induced nephrosis rats. To quantify the activation of the MAPK pathway, western blotting was used to detect the phosphorylation levels of MAPK pathway-associated proteins (p38, ERK1/2, SAPK/JNK) and nuclear factor (NF)-κB p65, which were reduced by the various treatments compared to those in the untreated doxorubicin-induced rats. Serum levels of transforming growth factor (TGF)-β1, tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6, quantified by ELISA, were decreased by the various treatments compared to the levels in the untreated doxorubicin-induced nephrosis rats. The rats treated with prednisone, simvastatin, and Shenkangling showed the best outcome. The Chinese medicine Shenkangling that is known for nourishing the kidney and promoting blood circulation reduced urinary protein levels, increased serum albumin levels, and reduced cholesterol levels by reducing the release of CXCL16, ADAM10, ADAM17, TGF-β1, TNF-α, IL-1β, IL- 6, and other inflammatory mediators and inhibiting the activation of the MAPK signaling pathway, thereby effectively improving the state of nephropathy in AN rats. These results indicate that Shenkangling can be used clinically to treat nephropathy.

Huaiqihuang Granules (槐杞黄颗粒) reduce proteinuria by enhancing nephrin expression and regulating necrosis factor κB signaling pathway in adriamycin-induced nephropathy

To investigate the effects of Huaiqihuang Granules (, HQH), a mixture of Chinese herbs including Trametes robiniophila Murr, Fructus Lycii and Polygonatum sibiricum, on adriamycininduced nephropathy (ADRN) in rats and its underlying mechanisms. Rats with ADRN were divided into four groups: the sham group, the model group (distilled water), the low-dose HQH-treated (2 g/kg) group, and the high-dose HQH-treated (4 g/kg) group. Body weight and 24-h urinary protein (Upro) were checked every week. After 5-week intervention, at the end of the study, the rats were sacrificed and blood samples were collected for examination of biochemical parameters, including glomerular morphological makers, podocyte shape, cellular apoptosis, expressions of nephrin, inflammatory and apoptosis markers. HQH ameliorated the rat's general status, proteinuria, renal morphological appearance and glomerulosclerosis. The decreased expression of nephrin in ADRN rats was increased by HQH, as well as the impaired podocyte foot process fusion. Cytosolic levels of p65 and inhibitor of nuclear factor κBα (IκBα) were decreased in ADRN rats, and recovered by the treatment of HQH. Consistently, the induced expression of tumor necrosis factor α (TNF-α), phosphorylated nuclear factor κB p65 (p-NFκB p65) and IκBα in ADRN were markedly suppressed by HQH. In addition, induction of Bax, cleaved caspase-3 and cytochrome C in ADRN rats were suppressed by HQH, indicating the amelioration of apoptosis. HQH could ameliorate renal impairments in ADRN rats by increasing nephrin expression, inhibiting NF-κB signaling pathway via the down-regulation of p-NF-κB p65 and p-IκBα, and suppression of glomerular and tubular apoptosis.

Astragaloside IV liposomes ameliorates adriamycin-induced nephritic syndrome in rats

Background : Radix Astragali was one of the main compositions of ‘Modified Danggui Buxue Decoction’  used for treatment of various kidney diseases. Astragaloside IV was the active composition of Radix Astragali . Astragaloside IV liposomes were used for the treatment of adriamycin-induced nephropathy (AN) rats. The aim of the study was to study the effect of astragaloside IV liposomes on AN rats, and to test through  regulating the expression of nephrin, integrins and integrin-linked kinase (ILK) in renal  tissues. Methods : The rats were given a single tail intravenous injection of adriamycin (6 mg/kg) within 1 week,  and then divided into four groups including normal, model, benazepril and astragaloside IV liposomes group. They were all orally administered dosage of benazepril and astragaloside IV liposomes once daily for 8 weeks. Results : Astragaloside IV liposomes significantly reduced the proteinuria of AN rats at 28, 42 and 56 days. astragaloside IV liposomes could increase the mRNA and protein expression of nephrin, integrin α3, down-regulate the expression of ILK to alleviate the podocyte damage and restore glomerular selective filtration function. Conclusions : Astragaloside IV liposomes could enhance renal function and protect podocyte to  ameliorate the adriamycin-induced nephritic syndrome. Key words ：Astragaloside IV, Liposomes, Ardiamycin-induce nephropathy, Podocyte

ASTRAGALOSIDE IV LIPOSOMES AMELIORATES ADRIAMYCIN-INDUCED NEPHRITIC SYNDROME IN RATS MING-GANG

Background: Radix Astragali was one of the main composition of ‘Modified Danggui Buxue Decoction’ used for treatment of various kidney&#x0D; diseases. Astragaloside IV was the active composition of Radix Astragali. Astragaloside IV liposomes were used for the treatment of&#x0D; adriamycin-induced nephropathy (AN) rats. The aim of the study was to study the effect of astragaloside IV liposomes on AN rats, and to test&#x0D; through regulating the expression of nephrin, integrins and integrin-linked kinase (ILK) in renal tissues.&#x0D; Methods: The rats were given a single tail intravenous injection of adriamycin (6 mg/kg) within 1 weeks, and then divided into four groups&#x0D; including normal, model, benazepril and astragaloside IV liposomes group. They were all orally administered dosage of benazepril and&#x0D; astragaloside IV liposomes once daily for 8 weeks.&#x0D; Results: Astragaloside IV liposomes significantly reduced the proteinuria of AN rats at 28, 42 and 56 days. astragaloside IV liposomes could&#x0D; increase the mRNA and protein expression of nephrin, integrin α3, down-regulate the expression of ILK to alleviate the podocyte damage and&#x0D; restore glomerular selective filtration function.&#x0D; Conclusions: Astragaloside IV liposomes could enhance renal function and protect podocyte to ameliorate the adriamycin-induced nephritic&#x0D; syndrome.

Effects of asiaticoside on levels of podocyte cytoskeletal proteins and renal slit diaphragm proteins in adriamycin-induced rat nephropathy

AimsCentella asiatica has been used to treat kidney diseases in Chinese traditional medicine. Asiaticoside (an extraction of C. asiatica) exerts a variety of pharmacological effects including immunomodulatory and anti-inflammatory functions. However, the mechanism of asiaticoside in the treatment of renal diseases remains largely unknown. This study investigated the molecular mechanism of asiaticoside in treating adriamycin-induced nephropathy of rats. Main methodsSixty-two SD male rats were randomly divided into normal control group (n=12) and nephropathy group (n=50). Except for the normal control group, rats were injected with adriamycin (6mg/kg) via the tail vein to induce nephropathy. Adriamycin induced nephropathic rats were divided into untreated group, prednisone group (25mg/kg), and asiaticoside groups with various dosages (8, 16 and 32mg/kg). Samples of urine and serum, tissue of kidney were collected for analysis after treatments for four weeks. Morphological changes were evaluated under light microscope and electron microscope. Synaptopodin, desmin, nephrin and podocin mRNA and protein were determined by RT-PCR and Western blotting. Key findingsCompared to the untreated nephropathy group, asiaticoside treatment mitigated histological damages, decreased 24-hour urine protein excretion and total cholesterol, increased serum albumin. Asiaticoside treatment increased the mRNA and protein levels of synaptopodin, nephrin and podocin in a dose-dependent manner. Furthermore, asiaticoside treatment decreased the mRNA and protein levels of desmin. SignificanceAsiaticoside can mitigate adriamycin-induced nephropathy in rats, which is associated with the increase in synaptopodin, nephrin and podocin gene expression, and the decrease in desmin gene expression.

Huangkui capsule, an extract from Abelmoschus manihot (L.) medic, ameliorates adriamycin-induced renal inflammation and glomerular injury via inhibiting p38MAPK signaling pathway activity in rats

Ethnopharmacological relevanceAbelmoschus manihot (L.) medic (AM) is a natural medicinal plant used for the treatment of inflammatory diseases in China. Huangkui capsule (HKC), an extract from AM, has been proved clinically effective in improving renal inflammation and glomerular injury in chronic kidney disease (CKD). However, the dose-effects and the mechanisms involved in vivo are still unclear. Aim of the studyThis study was performed to examine the dose-effects of HKC on renal inflammation and glomerular lesion in adriamycin-induced nephropathy (ADRN), then to clarify the mechanisms in vivo of HKC by investigating its actions on modulating the activation of p38 mitogen-activated protein kinase (p38MAPK) signaling pathway. Materials and methodsThe rats with chronic ADRN, created by the unilateral nephrectomy and twice adriamycin injections (ADR, 4mg/kg and 2mg/kg) within 4 weeks, were divided into four groups, a Sham group, a Vehicle group, a high-dose HKC group, and a low-dose HKC group, and that, sacrificed at the end of the 4th week after the administration. The rat's general status, renal morphological appearance, proteinuria, blood biochemical parameters, glomerular morphological changes, podocyte shape, and macrophage (ED1+ and ED3+ cells) infiltration in glomeruli were examined, respectively. The protein expressions of inflammatory cytokines including tumor necrosis factor (TNF)-α and interleukin (IL)-2, as well as p38MAPK signaling molecules such as transforming growth factor (TGF)-β1, p38MAPK, and phosphorylated-p38MAPK (p-p38MAPK), were also evaluated individually. ResultsHKC at high dose of 2g/kg/d not only significantly ameliorated the rat's general status, renal morphological appearance, proteinuria, albumin, and glomerulosclerosis, but also obviously reduced the infiltrated ED1+ and ED3+ macrophages in glomeruli and TNF-α protein expression in the kidney, in addition to these, evidently down-regulated TGF-β1 and p-p38MAPK protein expressions in ADRN rats, but had no influence on podocyte shape and renal function. ConclusionHKC could dose-dependently ameliorate renal inflammation and glomerular injury in ADRN rats, by way of reducing the infiltration and the activation of macrophages in glomeruli, and TNF-α protein expression in the kidney, as well as inhibiting p38MAPK signaling pathway activity via the down-regulation of p-p38MAPK and TGF-β1 protein expressions in vivo.

Podocyte injury and overexpression of vascular endothelial growth factor and transforming growth factor-beta 1 in adriamycin-induced nephropathy in rats

The aim of this study is to investigate the expression of nephrin, vascular endothelial growth factor (VEGF), transforming growth factor-beta 1 (TGF-β1), and podocyte number in adriamycin (ADR)-induced nephropathy. A total of 60 male Sprague–Dawley rats were randomly divided into the control group and the ADR nephropathy group. The nephropathy was induced by tail-vein injection of ADR (4mg/kg) twice at a 14-day interval. The expression levels of nephrin, VEGF, and TGF-β1 in glomeruli were assessed by immunohistochemistry and western blotting. The podocyte number was also evaluated after anti-Wilms’ tumor-1 (WT1) immunohistochemical staining. In addition, the urinary protein content, biochemical parameters in serum samples and glomerular sclerosis index (SI) were compared between groups. In the ADR nephropathy group, the expression levels of nephrin was significantly decreased with the fusion of podocyte foot processes at 6weeks after the first ADR injection, which was associated with a marked proteinuria. A decrease in podocyte number and an increase in SI with the overexpression of both VEGF and TGF-β1 were also observed in the glomeruli at 10weeks after the first ADR injection. This was associated with focal segmental glomerulosclerosis (FSGS). The study data suggest that podocyte injury and decreased nephrin, as well as increased VEGF and TGF-β1, may contribute to the development of proteinuria and FSGS in ADR-induced nephropathy in rats.

FK506 alleviates proteinuria in rats with adriamycin-induced nephropathy by down-regulating TRPC6 and CaN expression

To investigate the roles of transient receptor potential cation channel 6 (TRPC6) and calcineurin (CaN) in proteinuria pathogenesis and the mechanism of action of the calcineurin inhibitor tacrolimus (FK506) in adriamycin-induced nephropathy. The adriamycin-induced nephropathy rats were established and randomly divided into adriamycin nephropathy (ADR), low-dose FK506 treated (ADR + FK0.5), high-dose FK506 treated (ADR + FK1.0) and Control groups. Twenty-four hour urinary protein and blood biochemistry were measured on weeks 2, 3, 5 and 7, and the distributions and expressions of TRPC6 and CaN in the renal tissue were detected by immunohistochemistry, real-time PCR and Western blot. The study showed the 24-hour urinary protein increased significantly in ADR rats compared with Controls whilst for ADR rats hypoalbuminemia, hypercholesterolemia, renal functional lesion and renal pathologic changes appeared. The areas and intensities of TRPC6 and CaN expressed in the glomerulus and tubulointerstitium of ADR rats increased significantly. The expression of TRPC6 mRNA in ADR rats began to increase on the 3rd week and persisted to the 7th week, and CaN mRNA increased throughout the period. Protein expressions of TRPC6 and CaN in ADR rats were significantly higher than those of Controls. FK506 can inhibit CaN activity in the renal tissues and further decrease TRPC6 expression and therefore reduce renal damage and proteinuria in a dose-dependent manner. These findings suggested that TRPC6 and CaN were up-regulated on mRNA and protein levels in adriamycin-induced rats. FK506 had a therapeutic effect on the progression of proteinuria and renal damage by down-regulating of TRPC6 and CaN in the renal tissues.