Improvement of Na+-K+ ATPase Activity and Gene Expression Associated with Glomerular Ultrastructural Protection in Diabetic Nephropathy by Acacia senegal: Role of Oxidative Stress Disturbances / Mejora de la actividad Na+-K+ ATPasa y la Expresión Génica Asociada con la Protección Ultraestructural Glomerular en la Nefropatía Diabética por Acacia senegal: Papel de las Alteraciones del Estrés Oxidativo

SUMMARY: This study assessed the effects of Acacia Senegal (AS) combined with insulin on Na+/K+-ATPase (NKA) activity and mRNA expression, serum glucose, renal function, and oxidative stress in a rat model of diabetic nephropathy (DN). Sixty rats were equally divided into six groups: normal control, normal+AS, diabetic (DM), DM+insulin, DM+AS, and DM+insulin+AS groups. Diabetes mellitus (type 1) was induced by a single injection of streptozotocin (65 mg/kg), and insulin and AS treatments were carried until rats were culled at the end of week 12. Serum glucose and creatinine levels, hemoglobin A1c (HbA1c) were measured. Renal homogenate levels of NKA activity and gene expression, malondialdehyde, superoxide dismutase (SOD), catalase and reduced glutathione (GSH) were evaluated as well as kidney tissue histology and ultrastructure. Diabetes caused glomerular damage and modulation of blood and tissue levels of creatinine, glucose, HbA1c, malondialdehyde, NKA activity and gene expression, SOD, catalase and GSH, which were significantly (p<0.05) treated with AS, insulin, and insulin plus AS. However, AS+insulin treatments were more effective. In conclusion, combined administration of AS with insulin to rats with DN decreased NKA activity and gene expression as well as oxidative stress, and improved glycemic state and renal structure and function.

Este estudio evaluó los efectos de Acacia senegal (AS) combinada con insulina sobre la actividad Na+/K+- ATPasa (NKA) y la expresión de ARNm, la glucosa sérica, la función renal y el estrés oxidativo en un modelo de nefropatía diabética (ND) en ratas. Sesenta ratas se dividieron equitativamente en seis grupos: control normal, normal+AS, diabética (DM), DM+insulina, DM+AS y DM+insulina+AS. La diabetes mellitus (tipo 1) se indujo mediante una única inyección de estreptozotocina (65 mg/kg), y los tratamientos con insulina y AS se llevaron a cabo hasta que las ratas fueron sacrificadas al final de la semana 12. Se midieron niveles séricos de glucosa y creatinina, hemoglobina A1c (HbA1c). Se evaluaron los niveles de homogeneizado renal de actividad NKA y expresión génica, malondialdehído, superóxido dismutasa (SOD), catalasa y glutatión reducido (GSH), así como la histología y ultraestructura del tejido renal. La diabetes causó daño glomerular y modulación de los niveles sanguíneos y tisulares de creatinina, glucosa, HbA1c, malondialdehído, actividad y expresión génica de NKA, SOD, catalasa y GSH, los cuales fueron tratados significativamente (p<0,05) con AS, insulina e insulina más AS. Sin embargo, los tratamientos con AS+insulina fueron más efectivos. En conclusión, la administración combinada de AS con insulina a ratas con DN disminuyó la actividad de NKA y la expresión genética, así como el estrés oxidativo, y mejoró el estado glucémico y la estructura y función renal.

Mechanism of Dahuangtang pellets in regulating podocyte autophagy of diabetic nephropathy mice through AMPK/mTOR/ULK1 signaling pathway / 中国临床药理学与治疗学

AIM: To explore the intervention effect of Dahuangtang pellets (DHT) on diabetic nephropathy (DN) based on the AMP-activated protein kinase/mammalian target of rapamycin/unc-51-like kinase 1 (AMPK/mTOR/ULK1) signaling pathway. METHODS: Eight mice were randomly assigned to the model group, the dapagliflozin group, and the DHT (high, medium, and low dosage) group out of a total of 40 C57BL/KSJ-db/db (hereafter referred to as db/db) mice; another 10 C57BL/KSJ-db/dm mice were used as the normal group, saline was provided to the normal and model groups, and the mice in the treatment group received the appropriate medications. The medications were given for 10 consecutive weeks, once per day, to the mice in the treatment group. At weeks 0, 4, 8, and 10 of administration, fasting blood glucose (FBG) was assessed by drawing blood at a predetermined time from the tail vein; Urine samples were taken at 0, 5, and 10 weeks after treatment to evaluate the levels of albumin and creatinine, and the urinary albumin-creatinine ratio (ACR) was computed. After 10 weeks, mice in each group were assayed for 24 h total urine protein, serum creatinine (Scr), urea nitrogen (BUN) levels; Western blotting analysis was conducted to detect the expression of p-AMPK, p-mTOR, and p-ULK1, as well as the expression of autophagy related proteins homolog of yeast Atg6 (Beclin-1), autophagy-related proteins microtubule-associated protein 1 light chain 3 (LC3), P62 in renal tissue; Immunohistochemistry was used to measure the expression of podocyte lacunar membrane proteins (Nephrin, Podocin) in renal tissues; The pathological morphology of renal tissue was observed by light microscopy and transmission electron microscopy. RESULTS: Compared with the model group, FBG, ACR, and 24 h total urine protein were reduced in the dapagliflozin group and DHT groups of mice, and there was no statistically significant difference in Scr and BUN; In renal tissues, there is increased expression of p-AMPK and p-ULK1, decreased expression of p-mTOR, increased expression of LC3II / LC3I and Beclin-1, and decreased expression of P62 (P<0.01, P< 0.05); differentially upregulated in glomeruli are the podocyte lacunar membrane proteins Nephrin and Podocin (P<0.01, P<0.05); renal pathologic damage was reduced to varying degrees; transmission electron microscopy showed an increase in the number of autophagic vesicles and autophagic lysosomes. CONCLUSION: DHT can delay the development of DN by regulating the AMPK / mTOR / ULK1 signaling pathway, enhancing podocyte autophagy, and protecting glomeruli.

The protective effect and mechanism of cornuside on diabetic nephropathy model mice / 中国药房

OBJECTIVE To investigate the protective effect and potential mechanism of cornuside on diabetic nephropathy （DN） model mice. METHODS Male KK-Ay mice were fed with high-fat and high-sugar diet for two weeks to reproduce the DN model. The successfully modeled mice were randomly grouped into model group， aminoguanidine group （positive control，100 mg/kg） and cornuside group （100 mg/kg）， and male C57BL/6J mice were included as normal group， with 6 mice in each group. Administration groups were given relevant medicine intragastrically， and normal group and model group were given a constant volume of normal saline intragastrically， once a day， for 8 consecutive weeks. The levels of fasting blood glucose （FBG）， 24 h urinary protein， serum interleukin-12 （IL-12）， IL-10， blood urea nitrogen （BUN） and serum creatinine （Scr） were detected； the pathological injury， fibrotic change and glomerular microstructure of renal tissue were observed； the expressions of the receptor of advanced glycation end products （RAGE）， collagen type Ⅳ （COL-Ⅳ） and inducible nitric oxide synthase （iNOS） in renal cortex were detected in each group. RESULTS Compared with normal group， the renal cortex of mice in model group showed obvious inflammatory cell infiltration and fibrotic changes； the mesangial hyperplasia of glomerulus was serious and the basement membrane had a large number of irregular dark dense deposits； the levels of FBG and 24 h urinary protein， the serum levels of IL- 12， BUN and Scr， and the expression levels of RAGE， COL-Ⅳ and iNOS in the renal cortex were significantly increased， while the serum level of IL-10 was significantly decreased （P＜0.01）. Compared with the model group， the renal pathological injuries， fibrotic changes and glomerular microstructure of mice in administration groups were improved significantly， and the above quantitative indexes were generally improved （P＜0.05 or P＜0.01）. CONCLUSIONS Cornuside has a certain protective effect on DN model mice. It can inhibit the inflammatory response， reduce urinary protein excretion， and alleviate renal fibrosis， which may be related to the inhibition of the advanced glycation end products/RAGE signaling pathway.

Dendrobii Caulis Mixture-containing Serum Protects Mice from Podocyte Injury Induced by High Glucose / 中国实验方剂学杂志

ObjectiveTo investigate the effect and mechanism of Dendrobium mixture （DMix）-containing serum on high glucose-induced podocyte injury in mice. MethodThe MPC5 mouse glomerular podocytes were cultured in vitro， and the optimal glucose concentration for modeling， modeling time， and concentration of DMix-containing serum for administration were determined. The cells were classified into normal （5.5 mmol·L-1 glucose+10% blank serum）， model （30 mmol·L-1 glucose+10% blank serum）， DMix-containing serum （30 mmol·L-1 glucose+10% DMix-containing serum）， ferroptosis inhibitor （Fer-1， 30 mmol·L-1 glucose+10% blank serum+1 μmol·L-1 Fer-1） groups. The corresponding kits were used to measure the levels of Fe2+ and lactate dehydrogenase （LDH） in cells. Enzyme-linked immunosorbent assay was employed to determine the content of glutathione （GSH） and lipid peroxide （LPO） in cells. Fluorescence probe was used to measure the reactive oxygen species （ROS） level. Real-time fluorescence quantitative polymerase chain reaction and Western blotting were employed to determine the mRNA and protein levels， respectively， of Wilms' tumor-1 （WT-1）， desmin， long chain acyl-CoA synthase 4 （ACSL4）， and glutathione peroxidase 4 （GPX4） in podocytes. ResultCompared with the blank group， the intervention with 30 mmol·L-1 glucose for 48 h reduced podocyte viability （P<0.01）， and the 10% DMix-containing serum showed the most significant improvement in podocyte viability （P<0.01）. Compared with the normal group， the model group presented elevated levels of Fe2+， LDH， LPO， and ROS， lowered GSH level， up-regulated mRNA and protein levels of desmin and ACSL4， and down-regulated mRNA and protein levels of WT-1 and GPX4 （P<0.01）. Compared with the model group， the DMix-containing serum lowered the Fe2+， LDH， LPO， and ROS levels， elevated the GSH level， down-regulated the mRNA and protein levels of desmin and ACSL4， and up-regulated the mRNA and protein levels of WT-1 and GPX4 in podocytes （P<0.05， P<0.01）. ConclusionDMix-containing serum exerts a protective effect on high glucose-induced podocyte injury by inhibiting ferroptosis.

Inhibitory effects of Ginkgo biloba extract on renal inflammation in diabetic nephropathy model mice and its mechanism / 中国药房

OBJECTIVE To investigate the inhibitory effects of Ginkgo biloba extract （GBE） on renal inflammation in diabetic nephropathy （DN） model mice， and its potential mechanism. METHODS KK/Ay mice were fed with high fat and high sugar to induce DN model. They were divided into model group， positive control group [metformin 200 mg/（kg·d）]， GBE low-dose and high-dose groups [100， 200 mg/（kg·d）]， with 6 mice in each group. Six C57BL/6J mice were fed with a regular diet as the control group. Administration groups were given relevant liquid intragastrically， control group and model group were given constant volume of normal saline intragastrically， once a day， for 8 consecutive weeks. The body weight， fasting blood glucose， 24-hour food intake， 24-hour urine output， monocyte chemoattractant protein-1 （MCP-1）， interleukin-12 （IL-12）， IL-10， advanced glycation end products （AGEs）， blood urea nitrogen （BUN） and serum creatinine （Scr） of mice were measured， and the ratio of bilateral kidneys to body weight was also calculated. The pathological injury and fibrotic changes of the renal cortex were observed， and the expressions of macrophage polarization marker proteins [type M1： inducible nitric oxide synthase （iNOS）； type M2： arginase-1 （Arg-1）] and AGEs-the receptor of advanced glycation end products （RAGE）/Ras homolog gene pharm_chenjing@163.com family member A （RhoA）/Rho-associated coiled-coil forming protein kinase （ROCK） signaling pathway-related proteins were determined in renal cortex. RESULTS Compared with the model group， the symptoms such as renal cortical hyperplasia， vacuoles， infiltration of inflammatory cells， and renal cortical fibrosis had been improved in GBE low-dose and high-dose groups； body weight， serum level of IL-10， the expression of Arg-1 in the renal cortex were significantly higher than model group （P＜ 0.01）； fasting blood glucose， 24-hour food intake， 24-hour urine output， serum levels of MCP-1， IL-12， BUN， Scr and AGEs， the ratio of bilateral kidneys to body weight， renal injury score， the proportion of renal interstitial fibrosis， the protein expressions of iNOS， RAGE， RhoA and ROCK1 （except for GBE low-dose group） in renal cortex were significantly lower than model group （P＜0.01）. CONCLUSIONS GBE could improve kidney damage and alleviate inflammatory response in DN model mice， the mechanism of which may be related to inhibiting the AGEs-RAGE/RhoA/ROCK signaling pathway and regulating macrophage polarization.

Renal biopsy in diabetic patients: clinical indicators and pathological findings / Biopsia renal en pacientes diabéticos: indicadores clínicos y hallazgos patológicos

ABSTRACT Background: In addition to diabetic nephropathies (DNP), prevalence of nondiabetic nephropathies (NDNP) is also known to be frequent in patients diagnosed with type 2 Diabetes mellitus (DM). Early diagnosis of these conditions is important for the treatment and prognosis of these patients. Aim: This study aimed to investigate the relationships between clinical and laboratory findings of type 2 diabetic patients' renal biopsies. Material and Methods: We retrospectively reviewed the medical records of 140 patients who had diagnosis of type 2 DM and underwent renal biopsy from July 2020- August 2022 at nephrology clinics of Hospital Umraniye. Renal biopsy results, presence of hypertension, diabetic retinopathy, hematuria, proteinuria; duration of the disease, biopsy indications, glycated hemoglobin (HbA1c), serum creatinine, blood urea nitrogen, albumin, and proteinuria levels in 24h urine were measured. The statistical significance level was determined as p<0,05. Results: NDNP were detected in 43,7% of the patients. Among these the most common diagnosis was interstitial nephritis (20%). The most common biopsy indication was found to be nephrotic range proteinuria (30,7%). The difference between the DNP and NDNP patients' renal biopsy indications was statistically significant (p<0,001). DNP patients had a higher retinopathy incidence (60%,11%, p<0,001). A statistically significant difference was detected between the disease duration of DNP and NDNP groups (11,23 +5,74 years, p:0,002). According to multivariate regression analysis DR and HbA1c value, more than 7% have 4, 482 and 4,591-fold increased the risk of DNP incidence (p=0,021, p:0,024). Conclusion: Early diagnosis of DNP and NDNP of diabetic patients by performing renal biopsies affects the treatment and prognosis of the patients. Therefore, when evaluating diabetic patients, its necessary not to overlook the findings suggestive of NDNP.

RESUMEN Antecedentes: Además de las nefropatías diabéticas (DNP), también se conoce la prevalencia frecuente de nefropatías no diabéticas (NDNP) en pacientes diagnosticados con Diabetes mellitus tipo 2 (DM). El diagnóstico precoz de estas condiciones es importante para el tratamiento y pronóstico de estos pacientes. Objetivo: Este estudio tuvo como objetivo investigar las relaciones entre los hallazgos clínicos y de laboratorio de las biopsias renales de pacientes diabéticos tipo 2. Material y Métodos: Revisamos retrospectivamente las historias clínicas de 140 pacientes que tenían diagnóstico de DM tipo 2, desde julio de 2020 hasta agosto de 2022, y se les realizó biopsia renal en las clínicas de nefrología del Hospital Umraniye. Se revisaron los resultados de biopsia renal, presencia de hipertensión arterial, retinopatía diabética, hematuria y proteinuria así como también la duración de la enfermedad, las indicaciones de la biopsia, la hemoglobina glucosilada (HbA1c), la creatinina sérica, el nitrógeno ureico en sangre, la albúmina y los niveles de proteinuria en orina de 24 h. El nivel de significación estadística se determinó como p<0,05. Resultados: se detectaron NDNP en el 43,7% de los pacientes. Entre estos, el diagnóstico más común fue la nefritis intersticial (20%). La indicación de biopsia más frecuente resultó ser la proteinuria en rango nefrótico (30,7%). La diferencia entre las indicaciones de biopsia renal de los pacientes DNP y NDNP fue estadísticamente significativa (p<0,001). Los pacientes con DNP tuvieron una mayor incidencia de retinopatía (60%, 11%, p<0,001). Se detectó una diferencia estadísticamente significativa entre la duración de la enfermedad de los grupos DNP y NDNP (11,23 +5,74 años, p:0,002). De acuerdo con el análisis de regresión multivariado, la presencia de DR y el valor de HbA1c en más del 7% tienen 4,482 y 4,591 veces mayor riesgo de incidencia de DNP (p = 0,021, p: 0,024). Conclusión: El diagnóstico precoz de DNP y NDNP de pacientes diabéticos mediante la realización de biopsias renales afecta el tratamiento y pronóstico de los pacientes. Por lo tanto, al evaluar pacientes diabéticos, es necesario no pasar por alto los hallazgos sugestivos de NDNP.

The role of pro-inflammatory cytokines – tumor necrosis factor-alpha and interleukin 6 – in the evolution of diabetic nephropathy: A comparative study

Background: Diabetes is highly prevalent and it is responsible for the increased financial burden on healthcare. Type II diabetes is a more prevalent form of diabetes. Uncontrolled and unsupervised type II diabetes may lead to various microvascular and macrovascular complications which are responsible for high morbidity and mortality. Diabetic nephropathy (DN) is a common complication characterized by the expansion of mesangial cells with thickening of the basement and nodular glomerulosis. TNF-alpha and IL-6 play an important role in causing detrimental changes leading to nephropathy. The study of the role of these inflammatory cytokines in patients with DN may help in the early diagnosis and management. Aims and Objectives: The objectives of this study were to compare the levels of proinflammatory cytokines, TNF-?, and IL-6 in the evolution of DN patients. Materials and Methods: The present study was conducted in the Department of Biochemistry, in collaboration with the Department of Medicine (Nephrology unit); Pt. B.D. Sharma, Post Graduate Institute of Medical Sciences, Rohtak after ethical clearance. Forty patients with DN (Stages 3, 4, and 5) and forty patients with diabetes mellitus without nephropathy were taken up for study after taking informed consent. Results: The mean serum TNF-? levels in cases was 33.05 ± 29.22 pg/mL and in controls was 17.67 ± 12.33 pg/mL. On the basis of unpaired t-test, the difference between the groups was statistically highly significant (P < 0.05). The mean serum interleukin-6 levels in cases was 24.92 ± 30.16 pg/mL (2.95–155.55 pg/mL) and in controls was 6.76 ± 5.82 pg/mL (2.22–35.42 pg/mL). On the basis of the t-test, the difference between the groups was statistically highly significant (P < 0.05). Conclusion: TNF-? and IL-6 may serve as potential biomarkers for patients with DN and also in the development of newer therapeutic modalities for the prevention and treatment of DN.

Hypomagnesemia � An overlooked parameter in type 2 diabetes mellitus

Background: Diabetes Mellitus refers to a group of common metabolic disorders that share the phenotype of Hyperglycemia. It is the leading cause of morbidity and mortality throughout the world with an estimated worldwide prevalence of 439 million by 2030 and 19% of world抯 DM patients are Indians. Magnesium is an important co-factor for various enzymes involved in Insulin secretion and is involved in sodium-potassium ATPase pump. 25%-38% of Type 2 DM patients had Hypomagnesemia, which has also contributed in developing microvascular complications such as Diabetic Retinopathy (DR) and Diabetic Nephropathy (DN). Various studies have suggested that Magnesium supplementation in Type 2 DM patients with Hypomagnesemia have shown beneficial effects on insulin sensitivity and glucose metabolism. Aim and objectives: To study the prevalence of Hypomagnesemia in Type 2 DM patients and to study the association of Hypomagnesemia with microvascular complications such as DR and DN. Materials and methods: It is a hospital based Observational study carried out in 2022 for a period of 1 year including 60 patients fulfilling the ADA criteria for diagnosing T2DM and patients with Diabetic Retinopathy and Diabetic Nephropathy, and excluding patients with Malabsorption, Chronic diarrhoea, Renal Failure on diuretic therapy, Sepsis, Pancreatitis. Serum Magnesium levels of 1.6 mg/dl � 2.6 mg/dl is considered as normal range. Serum Magnesium were measured using Xylidyl blue colorimetric method. Results: The Mean age of the patients in our study was 55.89 years. Among 60 patients diagnosed with Diabetes Mellitus, 42 patients had Hypomagnesemia, 18 patients had Normomagnesemia (p- value: <0.0001). Patients with an HbA1c levels > 7% had Hypomagnesemia when to compared to patients with HbA1c <7% with a significant p value of 0.009. Hypomagnesemia was also associated with Diabetic Retinopathy and Diabetic Nephropathy with a significant p-value of 0.013 and 0.009 respectively. Conclusion: In our study, it has shown that patients with uncontrolled T2DM had Hypomagnesemia, which is also associated with micro-vascular complications of T2DM such as DR and DN.

Research progress of diabetic renal fibrosis / 中国药理学通报

Diabetic nephropathy (DN) is one of the most common and serious microvascular complications in patients with diabetes mellitus. Diabetic renal fibrosis ( DRF) is a major pathological change in the development of DN. In recent years the incidence of renal fibrosis (RF) has remained high. For diabetic patients, RF may expose them to kidney transplantation or even death, which brings a great burden to themselves and their families. Therefore, learning the pathogenesis and the current treat ment status of DRF is crucial for the treatment of the disease and the development of new drugs. Here we review the general situa¬tion of DN, the general situation, molecular mechanism, and the treatment of DRF,looking forward to providing a reference for the research and treatment of DRF.

The inhibitory effect of Averrhoa carambola DMDD on high glucose-induced endoplasmic reticulum stress IRE1α pathway and inflammation in renal tubular epithelial cell HK-2 / 中国药理学通报

Aim To investigate the inhibition effect of 2-dodecyl-6-methoxycyclohexa-2, 5-diene-l, 4-dione ( DMDD) on renal tubular epithelial cell HK-2 endo¬plasmic reticulum stress and inflammatory responses induced by high glucose. Methods HK-2 cells were cultured in vitro and divided into normal group, high glucose group, endoplasmic reticulum stress inhibitor 4-PBA group (5 mmoL • L ) , DMDD high, medium and low dose groups (8,4,2 μmol • L

Metabolomics study of corn silk on renal tissue and urine of diabetic nephropathy rats / 中国药理学通报

Aim To investigate the mechanism of corn silk decoction on diabetic nephropathy (DN) rats using metabolomics technology. Methods DN rat model was established by feeding with high-sugar and high-fat diet, combined with intraperitoneal injection of low dose streptozotocin. Renal organ index, fasting blood glucose, albumin creatinine ratio, serum creatinine, blood urea nitrogen, triglyceride, total cholesterol, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol indexes were measured, and the pathological changes of renal tissues were also observed to evaluate the intervention effect of corn silk on DN model rats. Further, UPLC/Q-TOF-MS technology was used to screen potential biomarkers in renal tissues and urine, combined with principal component analysis (PC A) and orthogonal partial least squares discriminant analysis (OPLS-DA). After identification by HM-DB and KEGG database, the biomarkers were imported into MetaboAnalyst for metabolic pathway analysis. Results All indexes and pathological damage of kidneys were improved in groups with different doses of corn silk, indicating that corn silk had a good intervention effect on DN. Metabolomic analysis showed that 18 biomarkers could be significantly called back by corn silk, and it involved 18 metabolic pathways mainly including phenylalanine, tyrosine and tryptophan biosynthesis, riboflavin metabolism, arachidonic acid metabolism, and tyrosine metabolism. Conclusions The mechanism of corn silk decoction intervention on DN may be related to amino acid metabolism, riboflavin metabolism, and arachidonic acid metabolism.

Regulation of Yes associated protein in diabetic nephropathy mice by Yishen Huashi Granule / 中国药理学通报

Aim To explore the protective effect of Yishen Huashi Granule (YSHS) on streptozotocin (STZ) - indueed diabetes nephropathy (DN) in mice and its possible mechanism. Methods The STZ induced DN mice model was established, which was randomly divided into model group, YSHS group and YAP inhibitor Vertepofin group, and the eontrol group was also established. The intervention was started eight weeks after the successful modeling with the course of four weeks. Urine protein concentration before and after intervention in each group as well as serum creatinine (Scr) and blood urea nitrogen (Bun) levels after intervention were measured. After the treatment, the mice were sacrificed, and the pathological changes of glomeruli were observed by light microscope HE staining. The protein expression of YAP, p-YAP S127 and CTGF were detected by Western blot, and the mRNA expressions of YAP, CTGF and podocyte functional proteins nephrin, podophyxin and WT1 were detected by RT-PCR. Results The biochemical indexes of YSHS group were better than those of model group, and HE staining showed that the pathological injury of glomeruli was improved. In the model group the protein expression of YAP, p-YAP (S127) and CTGF as well as the mRNA expression of YAP and CTGF increased, while the mRNA expression of nephrin, podocalyxin and WT1 decreased. After YSHS treatment, the protein expression of YAP, p-YAP S127, CTGF and the mRNA expression of YAP and CTGF decreased, while the mRNA expression of nephrin, podocalyxin and WT1 increased. Conclusions YSHS can reduce urinary protein, protect renal function and alleviate glomerular pathological injury in DN mice. Its possible mechanism may be related to the down-regulation of YAP in renal tissue, the reduction of CTGF expression level and the up-regulation of podocyte protein mRNA expression.

Effect of baicalin modulation of TLR4/MAPK/NF-ΚB signaling pathway on renal fibrosis in rats with diabetic nephropathy / 中国药理学通报

Aim To investigate the protective effect of baicalin on renal fibrosis in rats with diabetic nephrop- III (Col- III) athy (DN) and to investigate its mechanism of action. Methods A rat model of diabetic nephropathy was constructed. The rats were randomly divided into control group, model group, baicalin low dose group, baicalin medium dose group, baicalin high dose group and metformin group, with 10 rats in each group. Except for the control group, all rats in each group were fed with streptozotocin 65 mg • kg -

Mechanism of Zhenwu Decoction in improving renal inflammatory injury in mice with DN of spleen-kidney Yang deficiency syndrome by regulating ROCK/IKK/NF-κB pathway / 中国中药杂志

To investigate the intervention effect and mechanism of Zhenwu Decoction on diabetic nephropathy(DN) mice of spleen-kidney Yang deficiency syndrome based on the Rho-associated coiled-coil kinase(ROCK)/IκB kinase(IKK)/nuclear factor-κB(NF-κB) pathway. Ninety-five 7-week-old db/db male mice and 25 7-week-old db/m male mice were fed adaptively for one week. The DN model of spleen-kidney Yang deficiency syndrome was induced by Dahuang Decoction combined with hydrocortisone by gavage, and then the model was evaluated. After modeling, they were randomly divided into a model group, high-dose, medium-dose, and low-dose Zhenwu Decoction groups(33.8, 16.9, and 8.45 g·kg~(-1)·d~(-1)), and an irbesartan group(25 mg·kg~(-1)·d~(-1)), with at least 15 animals in each group. The intervention lasted for eight weeks. After the intervention, body weight and food intake were measured. Serum crea-tinine(Scr), blood urea nitrogen(BUN), fasting blood glucose(FBG), urinary albumin(uALb), and urine creatinine(Ucr) were determined. The uALb/Ucr ratio(ACR) and 24 h urinary protein(UTP) were calculated. Renal pathological morphology was evaluated by HE staining and Masson staining. The levels of key molecular proteins in the ROCK/IKK/NF-κB pathway were detected by Western blot. Enzyme-linked immunosorbent assay(ELISA) was used to detect interleukin-1β(IL-1β), interleukin-6(IL-6), interleukin-8(IL-8), interleukin-10(IL-10), and tumor necrosis factor-α(TNF-α). Compared with the blank group, the model group showed increased content of BUN, uALb, and SCr, increased values of 24 h UTP and ACR, decreased content of Ucr(P<0.05), enlarged glomeruli, thickened basement membrane, mesangial matrix proliferation, inflammatory cell infiltration, and collagen fiber deposition. The protein expression of ROCK1, ROCK2, IKK, NF-κB, phosphorylated IKK(p-IKK), phosphorylated NF-κB(p-NF-κB), and phosphorylated inhibitor of NF-κB(p-IκB) increased(P<0.05), while the protein expression of inhibitor of NF-κB(IκB) decreased(P<0.05). The levels of inflammatory factors IL-1β, IL-6, IL-8, and TNF-α increased(P<0.05), while the level of IL-10 decreased(P<0.05). Compared with the model group, the groups with drug treatment showed decreased levels of BUN, uALb, SCr, 24 h UTP, and ACR, increased level of Ucr(P<0.05), and improved renal pathological status to varying degrees. The high-and medium-dose Zhenwu Decoction groups and the irbesartan group showed reduced protein expression of ROCK1, ROCK2, IKK, NF-κB, p-IKK, p-NF-κB, and p-IκB in the kidneys(P<0.05), increased protein expression of IκB(P<0.05), decreased levels of serum inflammatory factors IL-1β, IL-6, IL-8, and TNF-α(P<0.05), and increased level of IL-10(P<0.05). Zhenwu Decoction can significantly improve renal function and renal pathological damage in DN mice of spleen-kidney Yang deficiency syndrome, and its specific mechanism may be related to the inhibition of inflammatory response by down-regulating the expression of key molecules in the ROCK/IKK/NF-κB pathway in the kidney.

Effect and mechanism of tournefolic acid B on the improvement of glucose metabolism and renal function in diabetes nephropathy model mice / 中国药房

OBJECTIVE To explore the role and underlying mechanism of tournefolic acid B （TAB） on the improvement of glucose metabolism and renal function in diabetic nephropathy （DN） model mice. METHODS DN model mice were established by high-fat diet combined with streptozotocin， and then randomly divided into model group， positive control group （vitamin E， 20 mg/kg）， TAB low-dose， medium-dose and high-dose groups （1， 2， 4 mg/kg）， with 12 mice in each group； normal control group was given regular diet. Each group was given relevant medicine or normal saline intragastrically， once a day， for 4 consecutive weeks. The glucose metabolic function was estimated by fasting blood glucose， glucose tolerance test， insulin tolerance test and serum insulin concentration. The renal coefficients and biochemical indicators related to renal function [serum uric acid， blood urea nitrogen， creatinine levels， and ratio of urine microalbumin to creatinine] were detected in mice； the contents of biochemical indicators related to oxidative stress [superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px）， malondialdehyde （MDA）， 8-hydroxydeoxyguanosine （8-OHdG）] were determined in renal tissue of mice； the pathological morphology of renal tissue was observed； the expressions of extracellular matrix （ECM） deposition related factors [transforming growth factor β1 （TGF- β1）， fibronectin （Fn）， type Ⅳ collagen （Col Ⅳ）] and protein kinase B （Akt）/nuclear factor erythroid 2-related factor 2 （Nrf2）/heme oxygenase-1 （HO-1） pathway related proteins were determined in renal tissue of mice. RESULTS Compared with normal control group， fasting blood glucose， area under glucose tolerance curve， area under insulin tolerance curve， serum insulin content， the levels of uric acid， urea nitrogen and creatinine @qq.com and ratio of urinary microalbumin to creatinine in serum， the contents of MDA and 8-OHdG and the protein expressions of TGF-β1， Fn and Col Ⅳ were increased significantly in model group （P＜0.05）， while the contents of SOD， GSH-Px and the protein expressions of p-Akt， Nrf2， HO-1 in renal tissue were decreased significantly （P＜0.05）； the significant thickening of the basement membrane， accumulation of mesangial matrix， glomerulosclerosis and interstitial fibrosis of the renal tubules were all found. Compared with model group， above indexes of mice were all reversed significantly in TAB groups （P＜0.05）， and pathological changes were alleviated in a dose-dependent manner. CONCLUSIONS TAB can improve blood glucose metabolism and kidney function and alleviate renal tubulointerstitial fibrosis in DN model mice， the mechanism of which may be associated with activating the Akt/Nrf2/HO-1 signaling pathway and suppressing ECM deposition.

The Level of N-Carboxymethyllysine and C-Reactive Protein in Type 2 Diabetes Mellitus and it’s Association with HbA1c in Diabetic Nephropathy

@#Introduction: N-Carboxymethyllysine (CML) is involved in diabetic nephropathy (DN) via production of oxidative stress, growth factors and cytokines. C-reactive protein (CRP) is an inflammatory marker associated with diabetes risk. This study is to determine the level of serum CML and CRP in Type 2 diabetes mellitus (T2DM) patients and healthy subjects and to determine the correlation between CML and CRP with glycated haemoglobin (HbA1c) in T2DM patients. Methods: This is a case-control study on 73 T2DM patients without nephropathy, 74 T2DM patients with nephropathy and 73 healthy subjects, aged from 18 to 65 years old. Fasting venous blood was taken and analysed for CML, CRP, HbA1c, and creatinine. The comparisons of serum CML and CRP among the three groups and the correlation between CML and CRP with HbA1c (in T2DM patients) were determined. Results: The differences in CML [median (Interquartile Range) (IQR)] between healthy subjects [131.80 (73.56) ng/ml] and T2DM patients without nephropathy [188.80 (55.95) ng/ml]; between healthy subjects and T2DM patients with nephropathy [237.70 (439.04) ng/ml] were statistically significant (P<0.001). The differences in CRP [median (IQR)] between healthy subjects [1.64 (1.91) ng/ml] and T2DM patients without nephropathy [2.15 (5.64) ng/ml]; between healthy subjects and T2DM patients with nephropathy [4.75 (6.91) ng/ml] were statistically significant (P<0.001). Logistic regression showed CML and CRP are independent predictors of diabetic groups. There was no correlation between HbA1c with CML and CRP in T2DM groups. Conclusion: Since serum CML and CRP are independent predictors of DN, their levels can be used to identify high-risk diabetic patients prone to developing DN.

Chinese Medicine Regulates NLRP3 Inflammasomes to Treat Diabetic Nephropathy： A Review / 中国实验方剂学杂志

Diabetic nephropathy （DN）， a major cause of chronic kidney disease （CKD）， aggravates the prevalence of end-stage renal disease （ESRD） and threatens human health. The pathogenesis of DN is complex， in which inflammation is a key pathological link in the cascade injury. Therefore， the treatment targeting inflammation helps to delay the progression of DN. NOD-like receptor protein 3 （NLRP3）， a classical proteasome， acts as an inducer of innate immune responses. The activated NLRP3 inflammasomes produce and release inflammatory mediators to trigger pyroptosis and uncontrolled autophagy and mediate the stress signals promoting renal fibrosis， thus participating in the development and progression of DN. The NLRP3 inflammasome as a core site inducing inflammation is widely involved in DN progression and may be a novel target. The active components and compound prescriptions of Chinese medicines are increasingly applied in the prevention and treatment of DN. The latest studies have discovered that Chinese medicines can treat DN by regulating the activation of NLRP3 inflammasomes. Although studies have been conducted to explore the mechanism of Chinese medicines in the treatment of DN via NLRP3 inflammasome， the systematic review remains to be carried out. This paper reviews the relevant publications in recent years and introduces the research progress from the assembly and activation of NLRP3 inflammasomes， the mechanism of NLRP3 inflammasomes in the treatment of DN， and the regulation of NLRP3 inflammasomes by Chinese medicines for the prevention and treatment of DN， aiming to lay a foundation for the relevant studies and provide new targets and strategies for the prevention and treatment of DN.

Research progress on innovative drugs for diabetic nephropathy with potential anti-inflammatory targets / 药学实践杂志

Diabetic nephropathy (DN) is a common microvascular complication of type 1 diabetes mellitus (T1DM) and type 2 diabetes mellitus (T2DM)，which is also the main cause of chronic kidney disease (CKD) and end-stage renal disease (ESRD). However, the treatment methods are limited at present. More and more evidences have indicated that inflammatory response is involved in the pathogenesis and progression of DN. Several anti-inflammatory strategies that target specific inflammatory mediators (transcription factors, pro-inflammatory cytokines, chemokines, adhesion molecules) and intracellular signaling pathways have shown benefits in the DN rodent model. The mechanisms related to inflammation in the development and progression of DN were summarized and new strategies to prevent or treat DN based on inflammation were briefly discussed in this review.

Exosomal lncRNA ZEB1-AS1 secreted from mesenchymal stem cells attenuates diabetic nephropathy by regulating miR-142-5p/PTEN axis / 中华内分泌代谢杂志

Objective:To explore the role of an exosomal long non-coding RNA zinc finger E-box-bingding homeobox 1 antisense 1(ZEB1-AS1) from adipose-derived MSC(adMSC) and its action mechanism in DN.Methods:DN rat model and high glucose(HG)-induced glomerular mesangial cell(GMCs) model treated with exosomal ZEB1-AS1 from adMSC were used for biochemical analysis and inflammation/oxidative stress assessment. The binding relationships among ZEB1-AS1, microRNA(miR)-142-5p, and phosphatase and tensin homolog deleted on chromosome 10(PTEN) were confirmed by dual luciferase reporter assay. Western blotting was used for the measurement of PTEN protein level.Results:adMSC-secreted exosomal ZEB1-AS1 reduced the levels of blood glucose, serum creatinine, 24 h urinary protein, kidney weight, fibrosis, and inflammatory cell infiltrations in DN rats. Meanwhile, both in DN rats and HG-induced GMCs, the levels of tumor necrosis factor(TNF)-α, interleukin(IL)-6, and IL-1β were inhibited, but glutathione peroxidase(GPx), superoxide dismutase(SOD), catalase(CAT), and glutathione(GSH) were promoted by exosomal ZEB1-AS1 treatment. Additionally, miR-142-5p was identified to bind to ZEB1-AS1, while miR-142-5p further targeted PTEN. miR-142-5p overexpression or PTEN silencing reversed the inhibiting effects of exosomal ZEB1-AS1 on inflammatory cytokine levels and the promoting effects on the concentrations of antioxidant enzymes.Conclusion:Exosomal ZEB1-AS1 from adMSC prevents DN progression by controlling inflammation and oxidative stress via the miR-142-5p/PTEN pathway, suggesting that ZEB1-AS1 may serve as a potential and effective therapeutic target for treating DN.

Serum monocyte/high-density lipoprotein cholesterol ratio and urinary albumin/creatinine ratio in evaluating osteoporosis in diabetic nephropathy patients / 中华内分泌外科杂志

Objective:To investigate the value of serum monocyte/high-density lipoprotein cholesterol ratio (MHR) and urinary albumin/creatinine ratio (ACR) in the evaluation of osteoporosis in diabetic nephropathy patients.Methods:Diabetic nephropathy patients treated in Hangzhou Ninth People’s Hospital from Jun. 2019 to Jun. 2022 were selected. Gender, age, height and weight of all patients were collected and recorded, and body mass index (BMI) was calculated. Blood calcium (Ca), blood phosphorus (P), parathyroid hormone (PTH), monocyte count (M), high density lipoprotein (HDL-C), urinary microalbumin and creatinine were measured and recorded in all patients. MHR and ACR were calculated, MHR=M/HDL-C, ACR=urinary microalbumin/creatinine. Lumbar spine bone mineral density (L1-L4) was measured by dual-energy X-ray absorptiometry, which was divided into osteoporosis group and non-osteoporosis group.Results:Among the 117 diabetic nephropathy patients, 47 cases were osteoporotic and 70 cases were non-osteoporotic. The proportion of women in osteoporosis group was significantly higher than that in non-osteoporosis group, and BMI, PTH, MHR, ACR and bone mineral density were significantly higher than those in non-osteoporosis group, with statistical significance (all P<0.05). Multivariate binary Logistic regression analysis showed that female, MHR and ACR were independent risk factors for osteoporosis in diabetic nephropathy patients (all P<0.05). Spearman correlation analysis showed that serum MHR and ACR were negatively correlated with lumbar bone density, with statistical significance ( r=0.524 and 0.497, P=0.004 and 0.009, respectively). ROC curve analysis showed that the area under the curve (AUC) of serum MHR and ACR for evaluating osteoporosis in diabetic nephropathy patients was 0.870 (0.809-0.931) and 0.849 (0.792-0.905), respectively. The AUC of serum MHR combined with ACR for osteoporosis in diabetic nephropathy patients was 0.927 (0.891-0.964) . Conclusion:Serum MHR and ACR can be used as the evaluation indexes of osteoporosis in diabetic nephropathy patients, and their combined efficacy is better.