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.

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.

Traditional Chinese Medicine in Prevention and Treatment of Renal Fibrosis in Diabetic Nephropathy by Targeting Notch Signaling Pathway： A Review / 中国实验方剂学杂志

Diabetic nephropathy （DN） is one of the common chronic kidney diseases （CKD） worldwide and a major cause of end-stage renal disease （ESRD）， seriously threatening and affecting the life and health of the global population. Currently， the pathogenesis of DN is considered to be closely related to factors such as glucose metabolism disorders， abnormal lipid metabolism， oxidative stress， activation of inflammatory factors， autophagy， and cell apoptosis in the continuous high-glucose environment of the body. Renal fibrosis is an important pathological feature and ultimate pathological outcome of DN. Timely intervention in renal fibrosis is of significant clinical and practical importance for the prevention and treatment of DN. Due to the limitations of western medicine in treating DN， traditional Chinese medicine （TCM） intervention in the process of renal fibrosis in DN has been widely used as a routine and potential treatment method due to its multi-component， multi-effect， and multi-target effects， effectively delaying the progression of the disease. It has been found that the Notch signaling pathway plays an important role in the development and maintenance of homeostasis in the body， and abnormal activation of the Notch signaling pathway is associated with DN. Activation of this signaling pathway plays a key role in the process of renal fibrosis. This article reviewed the regulatory mechanism of the Notch signaling pathway in renal fibrosis in DN， focusing on the relationship between targeting Notch signaling pathway by Chinese medicinal monomers and prescriptions and renal fibrosis in DN in order to provide a theoretical basis for the development of new drugs， basic research， and clinical application of TCM in the prevention and treatment of DN.

Oxidative stress status and its correlation with UACR and Uβ2-MG levels in patients with diabetic nephropathy / 公共卫生与预防医学

Objective To analyze oxidative stress status and its correlation with urinary albumin creatinine ratio (UACR) and urinary β2 microglobulin (Uβ2-MG) in patients with diabetic nephropathy (DN), and to provide a theoretical basis for clinical evaluation of oxidative stress status in DN patients. Methods A total of 382 DN patients admitted to our hospital from January 2020 to December 2021 were selected. According to the 24h urinary microalbumin excretion rate (24h UAER), the patients were divided into mild renal injury group (20µg/min 2-MG levels in DN patients (r=-0.462, 0.413, P2-MG levels in DN patients (r=-0.438, -0.459, P2-MG to predict the oxidative stress status of DN patients was 0.689, the sensitivity was 79.84%, and the specificity was 83.45%. Conclusion Oxidative stress in DN patients can accelerate the pathological progression of nephropathy. The oxidative stress status is closely related to the levels of UACR and Uβ2-MG, which can be used to judge the oxidative stress of the body and prevent the pathological progression of nephropathy in DN patients.

Mechanism of Modified Shenqiwan in Relieving Renal Interstitial Fibrosis in Diabetic Mice Based on GSK-3β/CREB Pathway / 中国实验方剂学杂志

ObjectiveTo observe the effects of modified Shenqiwan on renal function and fibrosis in diabetic nephropathy mice and explore the underlying mechanism based on the glycogen synthase kinase-3β （GSK-3β）/cyclic adenosine monophosphate （cAMP） response element-binding protein （CREB） signaling pathway. MethodFifty male db/db mice and 10 db/m mice were used in this study. The fifty db/db mice were randomly divided into model group， irbesartan group， and low-， medium-， and high-dose modified Shenqiwan groups. The 10 db/m mice were assigned to the normal group. The mice in the low-， medium-， and high-dose modified Shenqiwan groups were administered with modified Shenqiwan in the dosage form of suspension of Chinese medicinal granules by gavage， those in the irbesartan group were given irbesartan suspension by gavage， and those in the normal and model groups were given distilled water of equal volume by gavage. The intervention lasted for 12 weeks. The blood glucose levels， urine albumin-to-creatinine ratio （UACR）， and the protein expression levels of GSK-3β， CREB， transforming growth factor-β1 （TGF-β1）， E-cadherin， Vimentin， fibronectin （FN）， plasminogen activator inhibitor-1 （PAI-1）， and Collagen type Ⅳ （Coll Ⅳ） in the mouse kidneys were recorded before and after treatment. The extent of renal pathological damage was also observed. ResultCompared with the normal group， the model group showed significant increases in blood glucose levels， UACR levels， and the protein expression levels of GSK-3β， TGF-β1， E-cadherin， Vimentin， FN， PAI-1， and Coll Ⅳ in the kidneys （P<0.05）， decreased protein expression level of CREB （P<0.05）， and severe renal pathological damage. Compared with the model group， the low-， medium-， and high-dose modified Shenqiwan groups and the irbesartan group showed varying degrees of decreases in blood glucose levels， UACR levels， and the protein expression levels of GSK-3β， TGF-β1， E-cadherin， Vimentin， FN， PAI-1， and Coll Ⅳ in the kidneys （P<0.05）， increased expression level of CREB protein （P<0.05）， and improved renal pathological damage. ConclusionModified Shenqiwan can effectively reduce blood glucose levels， improve renal function， and alleviate fibrosis， and the mechanism of action is related to the inhibition of the GSK-3β/CREB signaling pathway.

Research progress of active ingredients of traditional Chinese medicine in the intervention of diabetic nephropathy by regulating mTOR signaling pathway / 中国药房

Diabetic nephropathy （DN） has become one of the main causes of end-stage renal disease，and its pathogenesis is still unclear. Currently，it is believed to be closely related to kidney injury mediated by various factors such as autophagy disorder under the condition of high glucose，oxidative stress and inflammation. Mammalian target of rapamycin （mTOR） signaling pathway is crucial for protein synthesis and autophagy regulation，which plays an important role in the occurrence and development of DN. In recent years，the research on the prevention and treatment of DN with traditional Chinese medicine （TCM） has made important progress. Plenty of evidence has shown that the active ingredients of TCM can enhance autophagy，improve oxidative stress and inflammation，inhibit cell apoptosis and abnormal proliferation by regulating mTOR signaling pathway，so as to relieve pathological changes in the kidney such as podocyte injury，glomerular basement membrane thickening，mesangial tissue abnormalities and renal tubule dysfunction，thereby reducing proteinuria and improving renal function. All of the above are of great significance for delaying the progression of DN. This article systematically summarizes the research progress of saponins，flavonoids，polyphenols，alkaloids， terpenoids and other active ingredients of TCM intervening in DN through mTOR signaling pathway，in order to provide some reference for further basic research and the development of new drugs.

Intervention effect of Tongxinluo capsule on renal fibrosis in diabetic nephropathy rats / 中国药房

OBJECTIVE To explore the intervention effect and related mechanism of Tongxinluo capsule on renal fibrosis in rats with diabetic nephropathy （DN）. METHODS Eight rats were selected as control group （ordinary feed）， the remaining rats were given high-glucose and high-fat diet combined with ip injection of streptozotocin （35 mg/kg） to induce DN model. Model rats were randomly divided into model group （purified water）， irbesartan group （positive control， 14.12 mg/kg） and Tongxinluo capsule group （0.3 g/kg）， including 12 rats in the model group and 11 rats for each of the other two groups. All groups were given relevant medicine or water intragastrically， once a day， for 16 consecutive weeks. After the last medication， fasting blood glucose and 24 h urinary total protein （24 h UTP） were detected. Pathological changes in renal cortex of rats in each group were observed. Serum levels of tissue-type plasminogen activator （PA） and plasminogen activator inhibitor 1 （PAI-1） were measured. mRNA expressions of transforming growth factor-β（1 TGF-β1）， type Ⅳ collagen（COL-Ⅳ）， Wnt4 and β-catenin in renal cortex of rats were detected. The protein depositions or expressions of TGF-β1， COL-Ⅳ， focal adhesion kinase （FAK）， integrin-linked kinase （ILK）， E-cadherin， PA， PAI-1， Wnt4 and β-catenin in renal cortex of rats were observed or determined. RESULTS Compared with model group， 24 h UTP of rats in Tongxinluo capsule group were all significantly reduced （P＜0.05）； pathological damage and fibrosis of renal cortex were relieved； the expression of PA in serum and renal cortex was significantly increased， while PAI-1 level was significantly reduced （P＜0.05）； the depositions of COL-Ⅳ and TGF-β1 in renal cortex were all reduced， and corresponding mRNA expression was decreased significantly （P＜0.05）； the depositions of ILK and FAK were decreased， while the deposition of E-cadherin was increased； protein and mRNA expressions of Wnt4 and β-catenin were significantly reduced （P＜0.05）. CONCLUSIONS Tongxinluo capsule can relieve pathological damage to renal tissue and renal fibrosis of DN model rats， and reduce extracellular matrix deposition. The mechanism may be related to regulation of fibrinolytic system activity， the decrease of ILK and FAK expression， and inhibition of Wnt/β-catenin signaling pathway.

Effects of Intestinal Flora and Local Renin-angiotensin System on Diabetic Nephropathy and Traditional Chinese Medicine Intervention： A Review / 中国实验方剂学杂志

Diabetic nephropathy （DN） is a common clinical complication of diabetes， the main cause of end-stage renal disease （ESRD）， and a key determinant of survival in diabetic patients. The pathogenesis of DN is complex， and it is currently believed to be associated with hemodynamic abnormalities， intestinal flora disturbances， glucose and lipid metabolism disorders， oxidative stress， genetic susceptibility， and protein non-enzymatic glycosylation. The local renin-angiotensin system （RAS） has always been the core of the pathogenic and progressive changes of DN. Once activated， it will induce the massive release of oxygen free radicals in the blood vessels， damage the endothelial function， and affect the microcirculation of the body. The recent studies demonstrate that intestinal flora and its metabolites may affect the occurrence and development of DN by activating or antagonizing the local RAS. Compared with western medicine treatment， traditional Chinese medicine （TCM） has the advantages of multiple targets and little toxic and side effects. Many TCM scholars have found that single herbs， their active ingredient extracts， and TCM compound prescriptions can improve kidney function by regulating the local RAS or intestinal flora. Specifically， the Chinese medicinal materials tonifying spleen （Codonopsis Radix， Dioscoreae Rhizoma， Atractylodis Macrocephalae Rhizoma， and Poria）， replenishing kidney （Rehmanniae Radix Praeparata， Corni Fructus， and Pseudostellariae Radix）， and activating blood， resolving stasis， and dredging collaterals （Hirudo， Salviae Miltiorrhizae Radix et Rhizoma， and Angelicae Sinensis Radix） have the regulatory effect. This article summarizes the roles of intestinal flora and local RAS in the occurrence and development of DN， and analyzes the animal experiments or clinical trials of TCM intervention in DN in recent years， aiming to provide more therapies and a theoretical basis for the treatment of DN with integrated TCM and Western medicine.

Top 10 Academic Progress of Traditional Chinese Medicine in 2022 / 中国实验方剂学杂志

BackgroundIn order to implement the spirit of the 20th National Congress of the Communist Party of China， and the Opinions on Promoting the Inheritance， Innovation and Development of Traditional Chinese Medicine（TCM）， to regularly summarize the research results of TCM， to present the academic progress of TCM dynamically， and to give full play to the academic leadership of academic groups， the China Association of Chinese Medicine had organized the selection of the top 10 academic progress of TCM in 2022. The selection work adhered to the four orientations， eliminated the four only， highlighted the solution of clinical problems， answered scientific questions， led the development of the industry， reflected the exploratory and forward-looking， innovative and breakthrough， focused on new laws， new discoveries， new methods， new products， new theories in the field of basic research and applied basic research in TCM. After dynamic collection， preliminary examination， review and final judgment， the top 10 academic progress of TCM in 2022 were determined.

Traditional Chinese Medicine Treats Diabetic Nephropathy by Inhibiting Pyroptosis and Inflammasome： A Review / 中国实验方剂学杂志

Diabetic nephropathy （DN） is a serious complication of diabetes， a major risk factor for chronic renal failure and end-stage renal disease， a major cause of deaths of diabetic patients， and a major cause of noncommunicable disease-associated deaths in the world. Pyroptosis and inflammasome activation are closely associated with the occurrence and development of DN. They can mediate a variety of pathological changes such as the loss and fusion of podocytes， up-regulated expression of inflammatory cytokines， macrophage infiltration， glomerular mesangial matrix expansion， and increased urinary albumin-to-creatinine ratio， eventually leading to nephron loss and kidney injury. The available studies have reported that the active ingredients in Chinese medicinal herbs or classical prescriptions play a role in the treatment of DN by lowering blood glucose and lipid levels， mitigating insulin resistance， reducing inflammation， alleviating oxidative stress， and regulating immunity. Moreover， the active ingredients can inhibit the activation of inflammasomes and the pyroptosis of renal cells， repair the inflammation-induced damage， improve renal function， and slow the progression of DN， demonstrating definite therapeutic effect. Chinese medicines can treat DN in a multi-target， multi-pathway， and multi-system manner， possessing broad prospects in the prevention and treatment of DN. Despite the extensive studies about the traditional Chinese medicine （TCM） intervention of DN in vivo and in vitro， a comprehensive summary of experimental studies on the TCM intervention of DN model remains to be carried out. This paper reviews the research progress in pyroptosis， inflammasomes， roles of pyroptosis and inflammasomes in DN， and TCM intervention of DN， aiming to provide ideas and reference for the research and development of drugs for this disease.

Animal Modeling of Diabetic Nephropathy：A Study Based on Literature / 中国实验方剂学杂志

ObjectiveTo summarize the modeling elements， evaluation indicators， characteristics， and drawbacks of the animal models of diabetic nephropathy， and thus provide guidance for the standardized modeling and rational application of these models. MethodThe articles about the animal experiments of diabetic nephropathy published in the last decade were retrieved from China National Knowledge Infrastructure， Wanfang Data， and PubMed. The data of animal species， sex， modeling techniques， modeling criteria， and evaluation indicators were analyzed in Excel. ResultA total of 287 publications were included in this study. Male SD rats were mainly used for the modeling of diabetic nephropathy. The animal models of type 1 diabetes were mainly established by intraperitoneal injection of streptozotocin （STZ） at 60-69 mg·kg-1 once or 50 mg·kg-1 for 5 continuous days， and those of type 2 diabetes by intraperitoneal injection of STZ at 30-39 mg·kg-1 once or 30 mg·kg-1 for 2 continuous days combined with 4 weeks of high-fat and high-sugar diet. Blood glucose and 24-hour urine protein were mainly used to determine whether the modeling was successful. The evaluation indicators of the animal models mainly included basic indicators， glucose and lipid metabolism indicators， and renal function indicators. ConclusionAnimal models are commonly used in the research on diabetic nephropathy， while there is no unified standards for the preparation or evaluation of the animal models. Moreover， Chinese medicine is rarely considered in the modeling. Through literature review and data analysis， this paper summarizes the modeling elements and standards， key evaluation indicators， characteristics， and shortcomings， aiming to build the animal models of diabetic nephropathy with a high success rate and with the characteristics in line with the clinical pathogenesis and syndromes.

Mechanism of Hedysari Radix Polysaccharide on Diabetic Nephropathy in db/db Mice Based on JAK2/STAT3 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo observe the effect of Hedysari Radix polysaccharide （HRP） on the Janus kinase 2 （JAK2）/signal transducer and activator of transcription protein 3 （STAT3） signaling pathway in diabetic nephropathy db/db mice. MethodFifty db/db mice were randomly divided into model group， irbesartan group （irbesartan suspension， 22.75 mg·kg-1）， and high-， medium-， and low-dose HRP groups （HRP suspension， 200， 100， 50 mg·kg-1） according to the body weight， with 10 mice in each group. Another 10 C57BL/6 mice were assigned to the normal group. The mice were treated with corresponding drugs by gavage， while those in the normal group and the model group received distilled water at 5 mL·kg-1. The mice in the six groups were administered once a day by gavage for 12 consecutive weeks. The uric acid （UA）， triglycerides （TG）， and total cholesterol （TC） were detected. Periodic acid-Schiff （PAS） staining and Masson staining were used to observe the pathological changes in kidney tissues. Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） were used to detect the protein and mRNA expression levels of JAK2， STAT3， suppressor of cytokine signaling 3 （SOCS3）， and tumor necrosis factor-α （TNF-α） in the kidney. ResultAfter 12 weeks of treatment， compared with the normal group， the model group showed significant pathological ultrastructural changes in kidney tissues and increased UA， TG， and TC levels （P<0.01）. Compared with the model group， the high- and medium-dose HRP groups and the irbesartan group showed improvement in pathological ultrastructure of kidney tissues and reduced UA， TG， and TC levels （P<0.05， P<0.01）. Compared with the normal group， the model group showed a decrease in SOCS3 protein and mRNA expression levels and an increase in JAK2， STAT3， and TNF-α protein and mRNA expression levels （P<0.01）. Compared with the model group， the high- and medium-dose HRP groups and the irbesartan group showed an increase in SOCS3 protein and mRNA expression levels and a decrease in JAK2， STAT3， and TNF-α protein and mRNA expression levels （P<0.05， P<0.01）. ConclusionHRP can alleviate renal damage in diabetic nephropathy to a certain extent， and its mechanism may be related to the inhibition of the activation of the JAK2/STAT3 signaling pathway.

Effect of Tripterygium wilfordii Polyglycoside on Expression of NFAT2/COX-2 in Kidney Tissues of Rats with Diabetic Nephropathy / 中国实验方剂学杂志

ObjectiveTo explore the underlying mechanism of Tripterygium wilfordii polyglycoside tablets （TWPT） in the prevention and treatment of kidney injury in diabetic nephropathy （DN） through the nuclear factor of activated T-cells 2（NFAT2）/cyclooxygenase-2（COX-2） pathway. MethodForty-two male SD rats of SPF grade were selected and randomly divided into a normal group （n=8） and an experimental group （n=34） after one week of adaptive feeding. The rats in the normal group were fed conventionally. The DN model was established in rats of the experimental group by intraperitoneal injection of streptozotocin （STZ） following one week of feeding on a high-fat and high-glucose diet. After the death and failure cases during modeling were eliminated， the remaining 24 model rats were randomly divided into model group， valsartan （8.33 mg·kg-1·d-1） group， and TWPT （5 mg·kg-1·d-1） group. Rats in normal group and model group were given equal amounts of normal saline by gavage. After six weeks， body weight was measured and urine samples were collected. Blood samples were collected from the abdominal aorta， and then the rats were sacrificed for sampling. Biochemical indicators， such as serum blood urea nitrogen （BUN）， serum creatinine （SCr）， alanine aminotransferase （ALT）， blood lipid， blood glucose， and 24-hour urine total protein （24 h UTP）， were determined. Hematoxylin-eosin （HE） staining and Masson staining were used to observe the pathology of the kidney. Enzyme-linked immunosorbent assay （ELISA） was used to detect NFAT2 and COX-2 expression levels in the serum. Western blot and Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）were adopted to detect NFAT2， COX-2 protein and mRNA expression in kidney tissues， respectively. ResultCompared with the normal group， the model group showed elevated 24 h UTP， BUN， SCr， CHO， TG， and FBG， increased serum NFAT2 and COX-2 production and expression （P<0.01）， and elevated protein and mRNA expression of NFAT2 and COX-2 in kidney tissues （P<0.01）. In addition， the pathology of the kidney showed enlarged glomeruli， mild proliferation of mesangial cells， and widened mesangial stroma. Compared with the model group， the TWPT group showed decreased 24 h UTP， BUN， SCr， CHO， TG， and FBG （P<0.05，P<0.01）， basically normal glomerular morphology， decreased expression of serum NFAT2 and COX-2 （P<0.01）， and down-regulated protein and mRNA expression of NFAT2 and COX-2 in kidney tissues （P<0.01）. ConclusionTWPT can alleviate 24 h UTP in DN model rats， protect renal function， and improve renal pathology， and its mechanism of action may be related to the down-regulation of NFAT2/COX-2 expression in the serum and kidney tissues.

Association of epidemiological characteristics and dietary patterns with risk of type 2 diabetic nephropathy / 公共卫生与预防医学

Objective To study the prevalence of diabetic nephropathy (DN) and the relationship between dietary pattern and risk. Methods From August 2018 to May 2021, 655 T2DM patients in Wuhan Puren Hospital, including 338 males and 317 females, were divided into T2DM group (n=368 cases) and DN group (n=287 cases) according to whether patients had DN. The uniformly trained staff of our hospital used the simplified version of food intake frequency questionnaire designed for diabetes to investigate the reasonable dietary intake of patients in nearly one year. Clinical data of patients in the two groups were collected and the intake of protein, fat, carbohydrate and dietary fiber in the two groups was statistically analyzed. Logistics regression was used to analyze the formula independent risk factors of DN in T2DM, and Pearson correlation analysis was performed on the incidence of diabetic nephropathy and dietary pattern. Results Among of 655 T2DM patients, there were 287 (43.82%) patients with DN, including 149 males and 138 females. The average age, duration of diabetes, smoking in DN group were significantly higher than those in T2DM group (P0.05). Energy, protein intake, carbohydrate and fat intake in DN group were significantly higher than those in T2DM group (P<0.05). Multivariate logistic regression analysis showed that protein, carbohydrate and fat intake were independent risk factors for DEVELOPING DN in T2DM patients (P<0.05). According to Pearson correlation analysis, the risk of developing DN in T2DM patients was positively correlated with protein and fat intake (r=0.449 , 0.517, P<0.05). Conclusion PATIENTS with T2DM have a higher risk of DEVELOPING DN, which is closely related to dietary intake. Reasonable allocation of dietary intake can reduce the risk of developing 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.

Discussion on the mechanism of Coptidis Rhizoma- Puerariae Lobamle Radix on diabetic retinopathy and diabetic nephropathy "different diseases with the same treatment" based on network pharmacology and molecular docking / 国际中医中药杂志

Objective:To explore the mechanism of Coptidis Rhizoma- Puerariae Lobamle Radix on the treatment of diabetic retinopathy (DR) and diabetic nephropathy (DN) by means of network pharmacology. Methods:The TCMSP and UniProt databases were used to retrieve the active components and targets of Coptidis Rhizoma and Puerariae Lobamle Radix. GeneCards and OMIM databases were used to search for DR and DN genes, and the online tool Venny was used to obtain intersection targets. Cytoscape 3.8.2 software was used to construct a network diagram of "components-targets", and the STRING platform was used to construct a protein interaction (PPI) network. GO function and KEGG pathway enrichment analysis were carried out through the DAVID annotation database. Molecular docking verification was performed. Results:A total of 18 active components and 74 disease-drug intersection targets were screened out from Coptidis Rhizoma- Puerariae Lobamle Radix. GO functional enrichment analysis showed that intersection targets were mainly concentrated in biological processes such as inflammation and apoptosis, involving cellular components such as extracellular space, plasma membrane, and cytoplasm, and was related to molecular functions such as protein binding, ATP binding, and enzyme binding. Enrichment analysis of KEGG revealed that the intersection target may be related to TNF signaling pathway, Toll-like receptor signaling pathway, PI3K-Akt signaling pathway, etc. The results of molecular docking showed that the core component had a good binding energy with the core targets. Conclusion:Coptidis Rhizoma-Puerariae Lobamle Radix may regulate TNF signal pathway, Toll-like receptor signal pathway and PI3K/Akt signal pathway through TNF, IL6, TP53 and other targets, and play a role in inhibiting cell apoptosis, oxidative stress and reducing inflammation.