iridoids and Diabetic-Nephropathies

The global pandemic of diabetes and diabetes complications confers heavy pressure on public health. Novel antidiabetes strategies with negligible unwanted effects are urgently needed. Currently, the anti-hyperglycemic potential of plant-based functional ingredients has been explored to provide alternative strategies. As a kind of dietary bioactive compound, oleuropein has aroused the growing interest of researchers in diabetes and diabetes complications management. This review reveals the research progress of oleuropein in treating diabetes and diabetes complications and summarizes the molecular mechanisms involved in these beneficial effects of oleuropein. Oleuropein achieves amelioration of diabetes, the mechanisms of which include the modulation of insulin secretion, the repairment islet morphology, the activation of hepatic AMP-activated protein kinase singling, and the improvement of glucose tolerance and insulin resistance. Oleuropein also can relieve diabetes complications including diabetic nephropathy, diabetes cardiovascular complications, diabetic retinopathy, poor wound healing, diabetic neuropathy, and diabetic testicular dysfunction. Oleuropein reverses cell apoptosis, regenerates tissues, restores the histological organization, and decreases oxidative stress in treating diabetes complications. Taken together, oleuropein is a promising compound for diabetes and diabetes complications management and can be used as a nutraceutical to fight against these diseases.

Topics: Diabetes Complications; Diabetes Mellitus; Diabetic Nephropathies; Humans; Iridoid Glucosides; Iridoids; Oxidative Stress

This study aimed to investigate the effects of geniposide(GP) on the expression of prokineticin(PK2) and prokineticin receptor 1(PKR1) in db/db mice with diabetic nephropathy(DN), so as to explore how the PK2 signaling pathway participated in the pathological changes of DN and whether GP exerted the therapeutic effect through this signaling pathway. Male mice were randomly divided into four groups, namely db/m, db/db, db/db+GP, and db/m+GP groups, with five in each group. The mice in the db/db+GP and db/m+GP groups were gavaged with 150 mg·kg~(-1) GP for eight successive weeks. Afterwards, all the mice were sacrificed and the renal tissues were embedded. The morphological changes in glomerulus and renal tubules were observed by Masson and PAS staining. The expression levels of PK2, PKR1, and Wilm's Tumor Protein 1(WT_1) in podocytes were detected by immunohistochemistry, and the protein expression levels of PK2 and PKR1 in mouse kidney by Western blot. The morphological results showed serious glomerular and tubular fibrosis(Masson), high glomerular and tubular injury score(PAS), increased glomerular mesangial matrix, thickened basement membrane, exfoliated brush border of renal tubules, decreased WT_1 in glomerular podocytes, and massive loss of podocytes in the db/db group. After administration with GP, the glomerular and tubular fibrosis was alleviated, accompanied by improved glomerular basement membrane and renal tubule brush edge, and up-regulated WT_1. As revealed by further protein detection, in the db/db group, the expression levels of PK2 and PKR1 and p-Akt/Akt ratio declined, whereas the ratio of Bax/Bcl-2 rose. Ho-wever, PKR2 and p-ERK/ERK ratio did not change significantly. After administration with GP, the PK2 and PKR1 expression was elevated, and p-Akt/Akt ratio was increased. There was no obvious change in PKR2, Bax/Bcl-2 ratio, or p-ERK/ERK ratio. All these have demonstrated that GP improves the renal damage in DN mice, and PK2/PKR1 signaling pathway may be involved in such protection, which has provided reference for clinical treatment of DN with GP.

Topics: Animals; Diabetes Mellitus; Diabetic Nephropathies; Iridoids; Kidney; Male; Mice; Signal Transduction

The theory of inflammation is one of the important theories in the pathogenesis of diabetic nephropathy (DN). We herein aimed to explore whether loganin affected macrophage infiltration and activation upon diabetic nephropathy (DN) by a spontaneous DN mice and a co-culture system of glomerular mesangial cells (GMCs) and macrophage cells (RAW264.7) which was induced by advanced glycation end products (AGEs).. Loganin showed remarkable capacity on protecting renal from damage by mitigating diabetic symptoms, improving the histomorphology of the kidney, decreasing the expression of extracellular matrix such as FN, COL-IV and TGF-β, reversing the production of IL-12 and IL-10 and decreasing the number of infiltrating macrophages in the kidney. Moreover, loganin showed markedly effects by suppressing iNOS and CD16/32 expressions (M1 markers), increasing Arg-1 and CD206 expressions (M2 markers), which were the phenotypic transformation of macrophage. These effects may be attributed to the inhibition of the receptor for AGEs (RAGE) /monocyte chemotactic protein-1 (MCP-1)/CC chemokine receptor 2 (CCR2) signaling pathway, with significantly down-regulated expressions of RAGE, MCP-1 and CCR2 by loganin. Loganin further decreased MCP-1 secretion when RAGE was silenced, which means other target was involved in regulating the MCP-1 expression. While loganin combinated with the inhibitor of CCR2 exerted stronger anti-inhibition effects of iNOS expression, suggesting that CCR2 was the target of loganin in regulating the activation of macrophages.. Loganin could ameliorate DN kidney damage by inhibiting macrophage infiltration and activation via the MCP-1/CCR2 signaling pathway in DN.

Topics: Animals; Chemokine CCL2; Chemokine CCL8; Diabetes Mellitus, Experimental; Diabetic Nephropathies; Fibronectins; Glycation End Products, Advanced; Iridoids; Kidney; Macrophage Activation; Macrophages; Male; Mesangial Cells; Mice; Mice, Inbred C57BL; RAW 264.7 Cells; Receptors, CCR1; Receptors, CCR2; Signal Transduction; Transforming Growth Factor beta1

Diabetic nephropathy (DN) is a common pathological feature in patients with diabetes and the leading cause of end-stage renal disease. Although several pharmacological agents have been developed, the management of DN remains challenging. Geniposide, a natural compound has been reported for anti-inflammatory and anti-diabetic effects; however, its role in DN remains poorly understood. This study investigated the protective effects of geniposide on DN and its underlying mechanisms. We used a C57BL/6 mouse model of DN in combination with a high-fat diet and streptozotocin after unilateral nephrectomy and treated with geniposide by oral gavage for 5 weeks. Geniposide effectively improves DN-induced renal structural and functional abnormalities by reducing albuminuria, podocyte loss, glomerular and tubular injury, renal inflammation and interstitial fibrosis. These changes induced by geniposide were associated with an increase of AMPK activity to enhance ULK1-mediated autophagy response and a decrease of AKT activity to block oxidative stress, inflammation and fibrosis in diabetic kidney. In addition, geniposide increased the activities of PKA and GSK3β, possibly modulating AMPK and AKT pathways, efficiently improving renal dysfunction and ameliorating the progression of DN. Conclusively, geniposide enhances ULK1-mediated autophagy and reduces oxidative stress, inflammation and fibrosis, suggesting geniposide as a promising treatment for DN.

Topics: AMP-Activated Protein Kinases; Animals; Anti-Inflammatory Agents; Autophagy; Autophagy-Related Protein-1 Homolog; Cyclic AMP-Dependent Protein Kinases; Diabetes Complications; Diabetes Mellitus, Experimental; Diabetic Nephropathies; Diet, High-Fat; Disease Models, Animal; Fibrosis; Glycogen Synthase Kinase 3 beta; Iridoids; Mice; Mice, Inbred C57BL; Oxidative Stress

Genipin-1-β-d-gentiobioside (GG) is a kind of compound extracted from Gardenia jasminoides Ellis. The chemical structure of GG is similar to that of geniposide and has antidiabetic effects. We aimed to investigate the efficacy of GG on diabetic nephropathy (DN) in vivo and in vitro experiments and explore its potential mechanism.. For high-fat diet/streptozotocin-induced DN mice used in our study, the general features of mice were analysed after GG treatment. Oxidative stress parameters and inflammatory factors were also measured by commercial kits. Kidney damage was assessed using hematoxylin and eosin (H&E), periodic acid-Schiff (PAS) and Masson staining, respectively. In vitro, podocyte injury was assessed by TUNEL and flow cytometric analyses. AMP-activated protein kinase/silencing information regulator related enzyme 1 (AMPK/SIRT1)/nuclear factor-κB (NF-κB) pathway-related proteins were detected by AMPK-siRNA intervention and western blotting.. Treatment of GG could increase cell survival and attenuated kidney damage. Despite the presence of inflammatory and oxidative stress, when GG retained the expression of AMPK/SIRT1, it could be observed that the downstream NLRP3 inflammatory-related proteins were inhibited.. Results showed that the protective efficacy of GG on DN works together with hypoglycemia and suppressing oxidative stress and inflammation, which at least partly involved in APMK/SIRT1/NF-κB-dependent pathway.

Topics: AMP-Activated Protein Kinases; Animals; Diabetes Mellitus, Experimental; Diabetic Nephropathies; Gardenia; Inflammation; Iridoids; Kidney; Male; Mice, Inbred C57BL; NF-kappa B; NLR Family, Pyrin Domain-Containing 3 Protein; Oxidative Stress; Phytotherapy; Plant Extracts; Podocytes; Sirtuin 1

Rehmanniae Radix (RR) and Cornus officinalis (CO) are a typical herbal pair used to treat diabetic nephropathy (DN) in clinical practice. DN can be effectively treated by catalpol (Cat) and loganin (Log), the main active components of RR and CO respectively, through combating apoptosis, oxidative stress and inflammation. Herein, a spontaneous DN and podocyte injury model induced by advanced glycation end products (AGEs), i.e. KK-Ay mice, was used to explore the cooperative effects of Log and Cat on DN and the mechanism targeting the AGEs-RAGE (receptor for AGE) pathway.. Log and Cat alone or in combination mitigated diabetic symptoms, decreased the level of fasting blood glucose, and increased that of serum insulin. The two drugs alone or in combination protected renal function from damage, prevented extracellular matrix hyperplasia and glycogen deposition, as well as alleviated the loss of podocytes detected by histological assay and immunohistochemistry. Flow cytometry revealed that Log and Cat alone or in combination relieved the apoptosis of AGEs-induced podocytes in vitro. Silencing RAGE by RNA interference played a protective role in podocyte apoptosis, whereas overexpression of it worked oppositely. Western blot exhibited that Log and Cat alone or in combination inhibited the activation of RAGE/p38 MAPK/p65 NF-κB and RAGE/Nox4/p65 NF-κB pathways in podocytes. The inhibitory effects of drug combination were more evident than those of individual treatments.. Log and Cat cooperatively resisted the apoptosis of podocytes upon DN by targeting AGEs-RAGE and its downstream pathways p38 MAPK and Nox4.

Topics: Animals; Apoptosis; Diabetic Nephropathies; Drug Therapy, Combination; Glycation End Products, Advanced; Iridoid Glucosides; Iridoids; Male; Mice; Mice, Inbred C57BL; NADPH Oxidase 4; Oxidative Stress; p38 Mitogen-Activated Protein Kinases; Podocytes; Receptor for Advanced Glycation End Products; Signal Transduction

Geniposide (GE) can effectively inhibit diabetic nephropathy (DN), but its mechanism is unclear. The objective of this study was to explore the antidiabetic nephropathy effects of GE both in high fat diet/streptozotocin-induced DN mice and in high glucose-induced podocyte model. Renal function in DN mice was evaluated by levels of serum creatinine (Scr) and blood urea nitrogen (BUN). Renal inflammation was appraised by pro-inflammatory cytokines: Tumor necrosis factor α (TNF-α), Interleukin 6 (IL-6) and IL-1β via ELISA assay. Renal histopathology analysis was conducted via hematoxylin and eosin, Masson and periodic acid-silver metheramine staining. Cellular viability was measured by Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay. Moreover, the related proteins p-NF-κB, ASC, Cleave-IL-1β, NLRP3, Cleave-Caspase-1 and GSDMD-N in AMPK/SIRT1/NF-κB pathway were assayed by Western blotting. In order to further investigate the effects of GE on podocytes, we also assessed these protein levels in AMPK/SIRT1/NF-κB pathway after siRNA-AMPK intervention by Western blotting. GE alleviated renal dysfunction as evidenced by decreased levels of Scr, BUN, TNF-α, IL-6 and IL-1β. Histological examination revealed GE effectively attenuated kidney damage, including glomerular basement membrane thickening and inflammatory cells infiltration. AMPK, p-AMPK and SIRT1 levels were obviously decreased both in DN mice and in podocyte model, but GE reversed these changes. The protein expressions in APMK/SIRT1/NF-κB pathway were significantly decreased by GE treatment. These results suggested that GE could efficiently block oxidative stress and inflammatory responses accompanied with pyroptosis, thus inhibiting the development of DN, and its mechanism might be related to APMK/SIRT1/NF-κB pathway.

Topics: AMP-Activated Protein Kinases; Animals; Apoptosis; Blood Urea Nitrogen; Creatinine; Cytokines; Diabetes Mellitus, Experimental; Diabetic Nephropathies; Diet, High-Fat; Iridoids; Kidney; Male; Mice; Mice, Inbred C57BL; NF-kappa B; Podocytes; RNA, Small Interfering; Signal Transduction; Sirtuin 1

Renal pathology was a commonly seen complication in patients with diabetes. Geniposide (GPO) was previously demonstrated to modulate glucose metabolism in diabetes. This study was to investigate effects of GPO in streptozotocin-induced diabetic rats and its underlying mechanism. Renal function in diabetic rats was evaluated by levels of serum creatinine (Scr), blood urea nitrogen (BUN), and urinary albumin. Renal inflammation was appraised by inflammatory cells infiltration and pro-inflammatory cytokines production. Renal monocytes, T lymphocytes infiltration, and intercellular adhesion molecule-1 (ICAM-1) expression were quantitated by immunohistochemistry. Moreover, renal nuclear factor-kappa B (NF-κB) was assayed by Western blotting. Diabetic rats showed renal dysfunction as evidenced by increased levels of Scr, BUN, urinary albumin, and elevator renal index. Histological examination revealed significant glomerular basement membrane (GBM) thickening. However, GPO notably improved renal function and diabetes-induced GBM changes. Additionally, diabetic rats showed noteworthy renal inflammation,as reflected by enhancement of monocytes and T lymphocytes infiltration, increased expression of ICAM-1, tumor necrosis factor-α, interleukin-1 (IL-1), and IL-6. Interestingly, the level of monocytes infiltration positively correlated with the severity of GBM. Further study indicated diabetic rats displayed increased activation of NF-κB, indicated by increased expression of NF-κB p65, IKKα, and p-IκBα in renal tissue. However, all the changes in renal inflammation and NF-κB pathway were obviously reversed in GPO-treated diabetic rats. Our works indicate GPO ameliorates structural and functional abnormalities of kidney in diabetic rats, which is associated with its suppression of NF-κB-mediated inflammatory response.

Topics: Animals; Blotting, Western; Diabetes Mellitus, Experimental; Diabetic Nephropathies; Dose-Response Relationship, Drug; Hypoglycemic Agents; Iridoids; Kidney; Kidney Function Tests; Male; Metformin; NF-kappa B; Rats, Sprague-Dawley; Signal Transduction; Streptozocin

Diabetic nephropathy is the most common cause of end-stage renal disease in patients with diabetes. Advanced glycation end-products (AGEs) play a prominent role in the development of diabetic nephropathy. We herein evaluated the effects of loganin on diabetic nephropathy in vivo.. We established a diabetic nephropathy model in C57BL/6J mice with diabetes induced by streptozotocin and fed with diets containing high level of AGEs. Diabetic symptoms, renal functions, and pathohistology of pancreas and kidney were evaluated. AGE-RAGE pathway and oxidative stress parameters were determined.. The model mice exhibited characteristic symptoms of diabetes including weight loss, polydipsia, polyphagia, polyuria, elevated blood glucose levels and low serum insulin levels during the experiments. However, loganin at doses of 0.02 and 0.1g/kg effectively improved these diabetic symptoms. Loganin reduced kidney/body weight ratio, 24h urine protein levels, and serum levels of urea nitrogen and creatinine in diabetic mice to different degrees compared to positive controls. Moreover, loganin improved the histology of pancreas and kidney, and alleviated the structural alterations in endothelial cells, mesangial cells and podocytes in renal cortex. Finally, we found that loganin reduced AGE levels in serum and kidney and downregulated mRNA and protein expression of receptors for AGEs in kidney in diabetic mice. Loganin also reduced the levels of malondialdehyde and increased the levels of superoxide dismutase in serum and kidney.. Loganin improved diabetic nephropathy in vivo associated with inhibition of AGE pathways, and could be a promising remedy for diabetic nephropathy.

Topics: Analysis of Variance; Animals; Blood Urea Nitrogen; Blotting, Western; Diabetes Mellitus, Experimental; Diabetic Nephropathies; Disease Models, Animal; DNA Primers; Dose-Response Relationship, Drug; Gene Expression Regulation; Glycation End Products, Advanced; Iridoids; Kidney; Malondialdehyde; Mice; Mice, Inbred C57BL; Microscopy, Electron, Transmission; Molecular Structure; Oxidative Stress; Pancreas; Proteinuria; Receptor for Advanced Glycation End Products; Receptors, Immunologic; Reverse Transcriptase Polymerase Chain Reaction; Superoxide Dismutase

Podocyte injuries are associated with progression of diabetic nephropathy (DN). Apelin, an adipocyte-derived peptide, has been reported to be a promoting factor for DN. In this study, we aim to determine whether apelin promotes progression of DN by inducing podocyte dysfunction. kk-Ay mice were used as models for DN. Apelin and its antagonist, F13A were intraperitoneally administered for 4 weeks, respectively. Renal function and foot process proteins were analysed to evaluate the effects of apelin on kk-Ay mice and podocytes. Apelin increased albuminuria and decreased podocyte foot process proteins expression in kk-Ay mice, which is consistent with the results that apelin receptor (APLNR) levels increased in glomeruli of patients or mice with DN. In cultured podocytes, high glucose increased APLNR expression and apelin administration was associated with increased permeability and decreased foot process proteins levels. All these dysfunctions were associated with decreased 26S proteasome activities and increased polyubiquitinated proteins in both kk-Ay mice and cultured podocytes, as demonstrated by 26S proteasome activation with cyclic adenosine monophosphate (cAMP) or oleuropein. These effects seemed to be related to endoplasmic reticulum (ER) stress, as apelin increased C/EBP homologous protein (CHOP) and peiFα levels while cAMP or oleuropein reduced it in high glucose and apelin treated podocytes. These results suggest that apelin induces podocyte dysfunction in DN through ER stress which was induced by decreased proteasome activities in podocytes.

Topics: Albumins; Animals; Apelin Receptors; Basement Membrane; Cell Membrane Permeability; Creatinine; Cyclic AMP; Diabetes Mellitus, Type 2; Diabetic Nephropathies; Endoplasmic Reticulum Stress; Female; Glucose; Humans; Intercellular Signaling Peptides and Proteins; Iridoid Glucosides; Iridoids; Kidney; Kidney Function Tests; Male; Mice, Inbred C57BL; Middle Aged; Podocytes; Proteasome Endopeptidase Complex; Proteasome Inhibitors; Receptors, G-Protein-Coupled

Renal tubular epithelial cell injury is a major pathological event that contributes to the development of diabetic kidney disease (DKD). Uncoupling protein-2 (UCP2), a mitochondrial membrane protein, has been reported to participate in the regulation of reactive oxygen species (ROS) generation, which contributes to tubular cell apoptosis induced by hyperglycemia. In this study, we found that genipin, a UCP2 inhibitor, dramatically boosted oxidative stress, attenuated antioxidative capacity, and exacerbated cell apoptosis accompanied with caspase-3 activation in rat renal proximal tubular cells (NRK-52E) incubated with high glucose. The present study results suggest that manipulation of UCP2 could be important in the prevention of oxidative stress damage in renal tubular epithelial cells induced by hyperglycemia in vitro.

Topics: Animals; Apoptosis; Cells, Cultured; Diabetic Nephropathies; Down-Regulation; Ion Channels; Iridoids; Kidney Tubules, Proximal; Mitochondrial Proteins; Rats; Uncoupling Protein 2

Connective tissue growth factor (CTGF) plays a pathogenic role in diabetic nephropathy (DN). Loganin, an iridoid glucoside compound was isolated from Cornus officinalis Sieb. et Zucc. This study was conducted to investigate the efficacy of loganin on DN and to elucidate the potential mechanism. High glucose (HG) stimulated cultured human renal proximal tubular epithelial cells (HK-2) analyzed CTGF expression by Western blotting and investigated whether extracellular signal-regulated kinase (ERK) signaling pathway was involved. Streptozotocin (STZ)-induced experimental DN, randomized to receive intragastric (i.g.) of loganin. Renal tissue, blood and urine samples were collected to determine and analyze. In vitro study, loganin reduced CTGF excretion in HG-induced HK-2 cells through the ERK signaling pathway. In vivo study, I.g. of loganin 5 mg/kg or 10 mg/kg significantly ameliorated renal function and increased body weight. Meanwhile, loganin reduced renal CTGF expression by immunohistochemical staining, reduced serum levels of CTGF. Besides, there were no significant differences in blood sugar levels between the loganin groups compared to the STZ-treated group. Furthermore, loganin ameliorated renal pathology. These results suggested that loganin exerts an early renal protective role to DN. Inhibition of CTGF may be a potential target in DN therapy, which highlights the possibility of using loganin to treat DN.

Topics: Animals; Blood Glucose; Cell Line; Cell Proliferation; Connective Tissue Growth Factor; Cornus; Cystatin C; Diabetes Mellitus, Experimental; Diabetic Nephropathies; Epithelial Cells; Glucose; Humans; Immunohistochemistry; Iridoids; Kidney; Male; MAP Kinase Signaling System; Rats; Rats, Sprague-Dawley; Weight Gain

Diabetic nephropathy (DN) is one of the most common causes of end stage renal disease (ESRD) in China, which requires renal replacement therapy. Recent investigations have suggested an essential role of podocyte injury in the initial stage of DN. This study investigated the potential therapeutic role of genipin, an active extract from a traditional Chinese medicine, on progression of DN in diabetic mice induced by intraperitoneally injection of streptozocin (STZ). In diabetic mice, orally administration of genipin postponed the progression of DN, as demonstrated by ameliorating body weight loss and urine albumin leakage, attenuating glomerular basement membrane thickness, restoring the podocyte expression of podocin and WT1 in diabetic mice. The protective role of genipin on DN is probably through suppressing the up-regulation of mitochondrial uncoupling protein 2 (UCP2) in diabetic kidneys. Meanwhile, through inhibiting the up-regulation of UCP2, genipin restores podocin and WT1 expression in cultured podocytes and attenuates glucose-induced albumin leakage through podocytes monolayer. Therefore, these results revealed that genipin inhibited UCP2 expression and ameliorated podocyte injury in DN mice.

Topics: Administration, Oral; Albuminuria; Animals; Cholagogues and Choleretics; Diabetes Mellitus, Experimental; Diabetic Nephropathies; Endopeptidases; Gene Expression Regulation; Iridoids; Male; Medicine, Chinese Traditional; Mice; Podocytes; Ubiquitin-Specific Proteases

Diabetic nephropathy (DN) is one of the foremost causes of renal failure and a primary cause of diabetes mellitus related death. Previously, we have reported that aqueous extract of Enicostemma littorale has potential antidiabetic activity. In the present study, we have investigated the effect of aqueous extract of E. littorale 1 g/kg, p.o. and swertiamarin 50 mg/kg, p.o. daily for 3 weeks in type 1 DN complications in SD rats. DN was assessed by serum urea, creatinine, lipid profile and water intake levels. Treatment with aqueous extract of E. littorale and swertiamarin significantly decreased serum urea and creatinine and other parameters associated with the development of DN in type 1 diabetic rats. We have also found considerable improvement in histology of glomerular function of aqueous extract of E. littorale and swertiamarin-treated animals.

Topics: Administration, Oral; Animals; Biomarkers; Blood Glucose; Creatinine; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 1; Diabetic Nephropathies; Drinking; Gentianaceae; Glucosides; Hypertrophy; Hypoglycemic Agents; Iridoid Glucosides; Iridoids; Kidney Glomerulus; Lipids; Male; Plant Extracts; Pyrones; Rats; Rats, Sprague-Dawley; Urea

Advanced glycation end products (AGE) are involved in the alterations of renal mesangial cell (MCs) growth, a feature of early stages of diabetic nephropathy (DN). We postulate that morroniside and loganin, 2 components extracted from Cornus officinalis, may ameliorate the detrimental effects of AGE-induced MCs proliferation by preventing oxidative stress. Rat MCs cultured in AGE milieu were treated with morroniside and loganin. Results showed that morroniside and loganin inhibited AGE-induced MC proliferation as measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Fluorescence microscopy revealed that the morroniside and loganin improved the morphological changes of MCs. Flow cytometric analysis showed that morroniside and loganin inhibited the cell cycle of rat MCs. Furthermore, the level of reactive oxygen species was significantly reduced, and the activities of superoxide dismutase and glutathione peroxidase were markedly increased, whereas the level of malondialdehyde was not significantly reduced. These results suggest that morroniside and loganin regulate MC growth by preventing oxidative stress. Thus, this study provides a molecular basis for the use of morroniside and loganin in the early stages of DN.

Topics: Animals; Antioxidants; Cell Cycle; Cell Line; Cell Proliferation; Cell Shape; Cornus; Diabetic Nephropathies; Flow Cytometry; Glutathione Peroxidase; Glycation End Products, Advanced; Glycosides; Iridoids; Malondialdehyde; Mesangial Cells; Microscopy, Fluorescence; Oxidative Stress; Rats; Reactive Oxygen Species; Superoxide Dismutase