The Role of Marein During Endothelial-Mesenchymal Transition in Diabetic Kidney Disease / Rol de la Mareína Durante la Transición Endotelial-Mesenquimatosa en la Enfermedad Renal Diabética

SUMMARY: Marein is a flavonoid compound that reduces blood glucose and lipids and has a protective effect in diabetes. However, the effect and mechanism(s) of marein on renal endothelial-mesenchymal transition in diabetic kidney disease (DKD) have not been elucidated. In this study, single-cell sequencing data on DKD were analyzed using a bioinformation method, and the data underwent reduced dimension clustering. It was found that endothelial cells could be divided into five subclusters. The developmental sequence of the subclusters was 0, 1, 4, 2, and 3, of which subcluster 3 had the most interstitial phenotype.The expression of mesenchymal marker protein:Vimentin(VIM), Fibronectin(FN1), and fibroblast growth factor receptor 1 (FGFR1) increased with the conversion of subclusters. In db/db mice aged 13-14 weeks, which develop DKD complications after 8-12 weeks of age, marein reduced blood levels of glucose, creatinine, and urea nitrogen, improved structural damage in kidney tissue, and reduced collagen deposition and the expression of FN1 and VIM. Marein also up-regulated autophagy marker:Light chain 3II/I(LC3II/I) and decreased FGFR1 expression in renal tissue. In an endothelial-mesenchymal transition model, a high glucose level induced a phenotypic change in human umbilical vein endothelial cells. Marein decreased endothelial cell migration, improved endothelial cell morphology, and decreased the expression of VIM and FN1. The use of the FGFR1 inhibitor, AZD4547, and autophagy inhibitor, 3-Methyladenine(3-MA), further demonstrated the inhibitory effect of marein on high glucose-induced endothelial-mesenchymal transition by reducing FGFR1 expression and up-regulating the autophagy marker protein, LC3II/I. In conclusion, this study suggests that marein has a protective effect on renal endothelial- mesenchymal transition in DKD, which may be mediated by inducing autophagy and down-regulating FGFR1 expression.

La mareína es un compuesto flavonoide que reduce la glucosa y los lípidos en sangre y tiene un efecto protector en la diabetes. Sin embargo, no se han dilucidado el efecto y los mecanismos de la mareína sobre la transición endotelial- mesenquimatosa renal en la enfermedad renal diabética (ERD). En este estudio, los datos de secuenciación unicelular sobre DKD se analizaron utilizando un método de bioinformación y los datos se sometieron a una agrupación de dimensiones reducidas. Se descubrió que las células endoteliales podían dividirse en cinco subgrupos. La secuencia de desarrollo de los subgrupos fue 0, 1, 4, 2 y 3, de los cuales el subgrupo 3 tenía el fenotipo más intersticial. La expresión de la proteína marcadora mesenquimatosa: vimentina (VIM), fibronectina (FN1) y receptor del factor de crecimiento de fibroblastos. 1 (FGFR1) aumentó con la conversión de subgrupos. En ratones db/db de 13 a 14 semanas de edad, que desarrollan complicaciones de DKD después de las 8 a 12 semanas de edad, la mareína redujo los niveles sanguíneos de glucosa, creatinina y nitrógeno ureico, mejoró el daño estructural en el tejido renal y redujo la deposición y expresión de colágeno de FN1 y VIM. Marein también aumentó el marcador de autofagia: Cadena ligera 3II/I (LC3II/I) y disminuyó la expresión de FGFR1 en el tejido renal. En un modelo de transición endotelial-mesenquimal, un nivel alto de glucosa indujo un cambio fenotípico en las células endoteliales de la vena umbilical humana. Marein disminuyó la migración de células endoteliales, mejoró la morfología de las células endoteliales y disminuyó la expresión de VIM y FN1. El uso del inhibidor de FGFR1, AZD4547, y del inhibidor de la autofagia, 3-metiladenina (3-MA), demostró aún más el efecto inhibidor de la mareína en la transición endotelial-mesenquimal inducida por niveles altos de glucosa al reducir la expresión de FGFR1 y regular positivamente la proteína marcadora de autofagia. , LC3II/I. En conclusión, este estudio sugiere que la mareína tiene un efecto protector sobre la transición endotelial-mesenquimatosa renal en la ERC, que puede estar mediada por la inducción de autofagia y la regulación negativa de la expresión de FGFR1.

Signaling Pathways Related to Renal Interstitial Fibrosis in Diabetic Kidney Disease Regulated by Traditional Chinese Medicine： A Review / 中国实验方剂学杂志

Diabetic kidney disease （DKD） is a common microvascular complication of diabetics mellitus （DM） and the leading cause of end-stage renal disease （ESRD）. Renal interstitial fibrosis （RIF） is the primary pathological basis for DKD progression to ESRD， which significantly increases the mortality rate of DKD patients and burdens patients and society， and it is thus a clinical problem that needs to be solved urgently. The pathogenesis of RIF is complex and mainly associated with excessive deposition of extracellular matrix （ECM）， epithelial-mesenchymal transition （EMT）， oxidative stress， inflammation， and autophagy. Multiple signaling pathways such as transforming growth factor-β1/Smad （TGF-β1/Smad）， nuclear transcription factor-κB （NF-κB）， p38 mitogen-activated protein kinase （p38 MAPK）， secretory glycoprotein/β-catenin （Wnt/β-catenin）， mammalian target of rapamycin （mTOR）， Janus kinase/signal transducer and activator of transcription （JAK/STAT）， neurogenic site-gap homologous protein （Notch）， and nuclear factor E2-associated factor 2 （Nrf2） mediate the development of RIF， which are currently novel targets for DKD therapy. Due to the complexity of its pathogenesis， the current Western medical treatment mainly focuses on essential treatment to improve metabolism， which has poor efficacy and is difficult to prevent the progression of DKD， so it is significant to find new treatment methods clinically. In recent years， many studies have proved that traditional Chinese medicine can alleviate oxidative stress， inhibit inflammatory response， and regulate cellular autophagy by modulating relevant signaling pathways， so as to treat RIF in DKD， which has the advantages of multi-pathway， multi-targeting， multi-linking， and significant therapeutic efficacy. However， there is still a lack of relevant summary. By reviewing the latest research reports in China and abroad， this article examines the roles of the signaling pathways mentioned above in the occurrence and development of RIF in DKD and the recent research progress in the intervention of RIF in DKD by traditional Chinese medicine via these pathways， aiming to provide new ideas and references for further scientific research and clinical practice.

Influence of Didang Xianxiong Decoction on Apoptosis of Podocytes in Diabetes Rats Through PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo observe the protective effect of Didang Xianxiong decoction on the kidneys of diabetic rats， its regulation on the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway， and its influence on podocyte apoptosis and explore the mechanism of Didang Xianxiong decoction in improving diabetic nephropathy. MethodThe diabetic model was established by a single intraperitoneal injection of streptozotocin （STZ） solution of 55 mg·kg-1. The successfully replicated model rats were randomly divided into the model group， Didang Xianxiong decoction group （8.10 g·kg-1）， Xiao Xianxiongtang group （4.05 g·kg-1）， Didangtang group （4.05 g·kg-1）， and alagebrium （ALT-711） group （3 mg·kg-1）， with six rats in each group. In addition， six rats were included in the blank group. After continuous administration for eight weeks， hematoxylin-eosin （HE） staining was used to observe the pathological changes in rats' kidney tissue. Masson staining was used to observe the degree of collagen deposition. Periodic acid-Schiff （PAS） staining was used to observe basement membrane lesions， and immunohistochemistry was used to detect the expression of phosphorylation （p）-PI3K and p-Akt proteins in rats' kidney tissue. The terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL） method was used to detect podocyte apoptosis. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of PI3K and Akt in rats' kidney tissue. Western blot was used to detect the protein expression of PI3K， p-PI3K， Akt， p-Akt， B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， phosphorylation glycogen synthase kinase-3β （p-GSK-3β）， and Caspase-3 in the kidney tissue. ResultCompared with the normal group， the model group had compensatory expansion of glomeruli， proliferation of mesangial cells， a large amount of collagen deposition in the mesangial stroma， thickening of the basement membrane， decreased mRNA expression of PI3K and Akt， and inhibition of PI3K and Akt protein phosphorylation （P<0.01）. It also underwent enhanced apoptotic signaling， decreased expression of anti-apoptotic protein Bcl-2 （P<0.01）， and increased expression of Bax， p-GSK-3β， and Caspase-3 （P<0.01）. Compared with the model group， Didang Xianxiong decoction significantly improved kidney tissue pathology， increased mRNA expression of PI3K and Akt （P<0.01）， significantly up-regulated phosphorylation levels of PI3K and Akt proteins （P<0.01） and Bcl-2 expression （P<0.01）， downregulated the expression of Bax， p-GSK-3β， and Caspase-3 （P<0.01）， and weakened podocyte apoptotic signaling. ConclusionDidang Xianxiong decoction may promote the activation of the PI3K/Akt signaling pathway， inhibit podocyte apoptosis， and thus slow down the progression of diabetic nephropathy.

Study on the Distribution of Traditional Chinese Medicine Syndrome Types in Patients with Diabetic Kidney Disease at Stages Ⅲ and Ⅳ / 广州中医药大学学报

Objective To investigate the distribution of traditional Chinese medicine(TCM)syndrome types in diabetic kidney disease(DKD),and to explore the correlation between TCM syndrome types and laboratory indices,so as to provide an objective basis for the TCM syndrome differentiation and treatment of DKD.Methods Syndrome differentiation was carried out in the 157 patients with DKD at stages Ⅲ and Ⅳ,and then the distribution of the syndromes of deficiency in the origin and the syndromes of excess in the superficiality was explored.The levels of 24-hour urinary total protein(24hUTP),serum creatinine(Scr),blood urea nitrogen(UREA),plasma albumin(Alb),total cholesterol(TC),and triglyceride(TG)of the patients were detected,and then the relationship between the TCM syndrome types and the biochemical indexes was analyzed.Results(1)The distribution of the syndromes of deficiency in the origin in DKD patients at different stages showed that DKD patients at stage Ⅲ were mainly differentiated as yin deficiency and dryness-heat syndrome[58.57％(41/70)],qi and yin deficiency syndrome[28.57％(20/70)],yin and yang deficiency syndrome[10.00％(7/70)],and spleen and kidney qi deficiency syndrome[2.86％(2/70)];DKD patients at stage Ⅳ were mainly differentiated as yin deficiency and dryness-heat syndrome[40.23％(35/87)],qi and yin deficiency syndrome[29.89％(29/87)],spleen and kidney qi deficiency syndrome[18.39％(16/87)],and yin and yang deficiency syndrome[11.49％(10/87)].The differences in the distribution of the syndromes of deficiency in the origin among the DKD patients at different stages were statistically significant(P<0.05).However,with the progression of the disease,DKD patients at different stages in general showed a trend of the decrease in the proportion of yin deficiency and dryness-heat syndrome while the increase in the proportions of qi and yin deficiency syndrome,spleen and kidney qi deficiency syndrome,and yin and yang deficiency syndrome.(2)The distribution of the syndromes of excess in the superficiality in DKD patients at different stages showed that DKD patients at stage Ⅲ were mainly differentiated as damp-heat syndrome[54.29％(38/70)],phlegm-stasis syndrome[27.14％(19/70)],blood-stasis syndrome[10.00％(7/70)],and cold-damp syndrome[8.57％(6/70)];DKD patients at stage Ⅳ were mainly differentiated as damp-heat syndrome[44.83％(39/87)],phlegm-stasis syndrome[35.63％(31/87)],cold-damp syndrome[14.94％(13/87)],and blood-stasis syndrome[4.60％(4/87)].There were no significant differences in the distribution of the syndromes of excess in the superficiality among the DKD patients at different stages(P>0.05).(3)The analysis of relationship between TCM syndrome type and biochemical indexes showed that Scr and UREA levels of DKD patients with spleen and kidney qi deficiency syndrome were significantly higher than those of patients with yin deficiency and dryness-heat syndrome,and the differences were statistically significant(P<0.05);Scr and 24hUTP levels of DKD patients with cold-damp syndrome were significantly higher than those of patients with damp-heat syndrome,and the differences were statistically significant(P<0.05).Conclusion DKD patients at stages Ⅲ and Ⅳ are all predominantly suffering from yin deficiency and dryness-heat syndrome,and with the progression of the disease,the syndrome of yin deficiency and dryness-heat develops into qi and yin deficiency syndrome,spleen and kidney qi deficiency syndrome,and yin and yang deficiency syndrome sequentially.Pathogenic dampness and blood stasis are the main pathogenic factors of DKD.And Scr,UREA,and 24hUTP are correlated with the TCM syndrome types of DKD,which will be helpful for the differentiation of TCM syndrome types of DKD.

Research progress of hypoxia inducible factor 1α in the mechanism of diabetic kidney disease / 基础医学与临床

Hypoxia-inducible factor-1α(HIF-1α)is a nuclear transcription factor.Under high glucose and hypoxia conditions,the expression of HIF-1α is elevated,result in the increased expression of its downstream target genes VEGF,HO-1 and BNIP3,which affect angiogenesis,extra cellular matrix deposition,iron metabolism,and mito-phagy,participating in the occurrence and development of diabetic kidney disease(DKD).In addition,HIF-1α promotes inflammation and renal fibrosis by affecting the production of cytokines in DKD.

Mechanism of FTO-mediated and m6A-modified PRKD2 Inhibiting Podocyte Injury in Diabetic Kidney Disease through the SIRT1/HIF-1 Pathway / 现代检验医学杂志

Objective To explore the regulatory role of fat mass and obesity-associated protein(FTO)and serine-threonine kinase protein kinase D2(PRKD2)in progression of diabetic kidney disease(DKD)and its regulatory mechanisms.Methods DKD model in vitro was constructed by podocytes(MPC5 cells)treated with high glucose(HG,35 mmol/L glucose)for 24 h.HG-induced MPC5 cells were transfected with FTO overexpression vector(pcDNA-FTO)and PRKD2 overexpression vector(pcDNA-PRKD2),or empty vector.The overexpression efficiency of FTO and PRKD2 were detected with RT-qPCR.MeRIP was used to detect the N6-methyladenosine(m6A)modification level of PRKD2 mRNA.The activity of Caspase-3 and the secretion of IL-6,TNF-α and monocyte chemotactic protein-1(MCP-1)were detected by ELISA.Cell apoptosis rate was analyzed by flow cytometry.The protein levels of FTO and PRKD2,as well as the key proteins in SIRT1/HIF-1α pathway,were evaluated by Western blot.Pearson analysis was used to analyze the correlation between FTO levels and PRKD2 levels.Results Compared with the control group without HG-induction,the protein expression of FTO(0.51±0.04 vs 1.00±0.03)and PRKD2(0.45±0.03 vs 1.01±0.04)was significantly down-regulated in HG-induced podocytes,and the differences were statistically significant(t=13.17,16.76,all P＜0.001).FTO protein levels were positively correlated with PRKD2 protein levels in HG-induced podocytes(r2=0.705 1,P＜0.001).Compared with the vector group,the m6A levels of PRKD2 mRNA(0.56±0.09 vs 1.01±0.13)in the pcDNA-FTO group were decreased,and the mRNA levels of PRKD2(3.16±0.14 vs 1.03±0.02)were increased,with significant differences(t=51.37,11.82,all P＜0.001).Compared with the control group(IL-6:512.76±61.85 pg/ml,TNF-α:28.17±2.83 pg/ml,MCP-1:157.31±17.69 pg/ml)and the vector group(IL-6:498.41±87.51 pg/ml,TNF-α:26.35±5.47 pg/ml,MCP-1:165.52±16.87 pg/ml),the secretion of IL-6(301.86±21.85 pg/ml),TNF-α(11.06±4.12 pg/ml)and MCP-1(81.45±9.03 pg/ml)were significantly decreased in the pcDNA-PRKD2 group,and the differences were statistically significant(F=7.51,10.47,61.97,all P＜0.01).Compared with the control group(caspase-3 activity:689.65±79.5 U/L,cell apoptosis:22.31%±2.69%)and the vector group(Caspase-3 activity:715.91±113.58 U/L,cell apoptosis:21.07%±3.28%),Caspase-3 activity(437.64±104.76 U/L)and the rate of apoptosis(8.41%±3.15%)were significantly decreased in the pcDNA-PRKD2 group,and the differences were statistically significant(F=2.35,79.13,all P＜0.01).Compared with the control group(SIRT1:1.01±0.05,HIF-1α:1.03±0.07)and the vector group(SIRT1:0.97±0.05,HIF-1α:1.02±0.03),SIRT1 protein levels(3.51±0.15)were increased and HIF-1α protein levels(0.37±0.07)were decreased in the pcDNA-PRKD2 group,and the differences were statistically significant(F=31.54,8.31,all P＜0.01).Conclusion FTO-mediated and m6A-modified PRKD2 suppresses inflammation and apoptosis in HG-induced podocytes through the SIRT1/HIF-1 pathway.

Research progress on indicators for early diagnosis of diabetic kidney disease / 沈阳医学院学报

Diabetic kidney disease(DKD)is one of the most serious microvascular complications of diabetes and a common cause of end-stage renal disease.Early kidney injury lacks typical clinical symptoms and its onset is insidiously.The common clinical indicators of kidney injury are urine microalbumin,glomerular filtration rate,etc.Due to their lack of sensitivity and specificity,it is easy to cause missed diagnosis,misdiagnosis,and delay the disease.Therefore,it is of great significance to find more convenient,stable,and less invasive detection indicators for the early diagnosis of DKD.This paper reviews the research progress of biological indicators related to glomerular and tubular injury,oxidative stress,inflammatory response,microRNA and proteomics,which provides a certain reference value for the early diagnosis of clinical DKD.

Bone morphogenetic protein 7 attenuates renal fibrosis in diabetic kid-ney disease rats by down-regulating Ajuba / 中国病理生理杂志

AIM:Bone morphogenetic protein 7(BMP7)reduces the expression of Yes-related protein 1(YAP1)by down-regulating Ajuba level and decreasing extracellular matrix(ECM)deposition.This study aimed to inves-tigate the influence of these factors on modifying the degree of renal fibrosis in rats with diabetic nephropathy.METH-ODS:Eighteen Sprague-Dawley(SD)rats were randomly divided into three groups:the normal control(NC)group,the diabetes mellitus(DM)group,and the DM group treated with BMP7 overexpressing adeno-associated virus(DM+rAAV-BMP7).Each group consisted of six rats.Diabetic kidney disease(DKD)was established in the DM and DM+rAAV-BMP7 groups by injecting 55 mg/kg streptozotocin(STZ)via the tail vein.NRK-52E cells were divided into three groups:the normal glucose(NG)group,the high glucose(HG)group,and the high glucose group treated with recombinant hu-man BMP7(HG+rhBMP7)group.Pathological changes in renal tissues were observed using hematoxylin and eosin(HE)and Sirius red staining.Immunohistochemical staining was performed to examine the expression sites of Ajuba and YAP1 in the renal cortex.Western blot analysis was conducted to determine the expression levels of BMP7,Ajuba,YAP1,colla-gen type Ⅲ(Col-Ⅲ),and fibronectin(FN)in the rat renal cortex and NRK-52E cells.RT-qPCR was used to measure the mRNA levels of Ajuba and YAP1 in the rat renal cortex.RESULTS:Biochemical indices revealed significantly ele-vated levels of blood glucose,serum creatinine,triglycerides,total cholesterol,and 24-hour urinary protein in the DM group compared to the NC group(P<0.05).In the DM+rAAV-BMP7 group,the levels of serum creatinine,24-hour uri-nary protein,triglycerides,and total cholesterol were lower than those in the DM group(P<0.05).Pathological staining demonstrated that the renal interstitium of the DM group exhibited inflammatory cell infiltration,fibrous tissue,collagen fi-ber deposition,disordered renal tubule arrangement,atrophy,and vacuolar degeneration,which were ameliorated in the DM+rAAV-BMP7 group.Immunohistochemistry revealed that Ajuba and YAP1 were mainly expressed in the cytoplasm and nucleus,with high expression in the cytoplasm of the DM group,which was significantly decreased in the DM+rAAV-BMP7 group.Western blot results indicated that the protein levels of FN,Col-Ⅲ,Ajuba,and YAP1 were up-regulated in the DM and the HG groups(P<0.05),but significantly down-regulated in the DM+rAAV-BMP7 group(P<0.05).RT-qP-CR results demonstrated that the mRNA levels of Ajuba and YAP1 were higher in the DM group and significantly lower in the DM+rAAV-BMP7 group(P<0.05).CONCLUSION:The overexpression of BMP7 can ameliorate renal fibrosis in rats with DKD.This effect is likely mediated by the down-regulation of Ajuba,reduction of YAP1 expression,and subse-quent inhibition of ECM deposition.

Expression of interleukin-36 and its regulatory effect on monocyte function in patients with diabetic kidney disease / 中华内分泌代谢杂志

Objective:To analyze the level of interleukin-36(IL-36) family cytokines in peripheral blood, and explore the regulatory role of recombinant human IL-36α in monocyte function in patients with diabetic kidney disease(DKD).Methods:A total of 41 type 2 diabetes mellitus(T2DM) patients, 36 DKD patients, and 20 controls were consecutively enrolled. Plasma and peripheral blood mononuclear cells(PBMCs) were isolated. Enzyme-linked immunosorbent assay(ELISA) was used to measure plasma levels of IL-36α, IL-36β, IL-36γ, and IL-36 receptor antagonist(IL-36Ra). PBMCs were sorted, and real-time quantitative PCR was performed to detect the mRNA expression of IL-36 receptor subunits in monocytes. Monocytes were stimulated with recombinant IL-36α, and levels of cytotoxic molecules and cytokines in the culture supernatant were measured. Flow cytometry was used to assess the expressions of programmed death receptor-1(PD-1) and cytotoxic T-lymphocyte-associated protein 4(CTLA-4). Co-culture of monocytes with Vero cells was performed to evaluate monocyte cytotoxicity.Results:Plasma levels of IL-36α and IL-36β in the T2DM and DKD groups were significantly higher than those in the control group. The DKD group also showed higher plasma levels of IL-36α compared to the T2DM group( P<0.05). There were no significant differences in IL-36γ and IL-36Ra levels among the three groups( P>0.05). The mRNA expression of IL-36 receptor subunits in monocytes was comparable among the three groups( P>0.05). The DKD group had higher level of tumor necrosis factor-alpha(TNF-α) compared to the control and T2DM groups( P<0.05). The levels of PD-1 and CTLA-4 were lower in the DKD group than those in the control and T2DM groups( P<0.05). The proportion of monocyte-induced Vero cell death was significantly higher in the DKD group compared to the control and T2DM groups( P<0.05). After stimulation with recombinant human IL-36α, monocytes from DKD patients showed a significant increase in the secretion of granzyme B and TNF-α( P<0.05), as well as an increased proportion of monocyte-induced Vero cell death( P=0.024). Conclusion:In DKD patients, elevated IL-36α and granzyme B levels in monocytes enhance monocyte function.

Influencing factors for hemodialysis initiation in non-diabetic kidney disease patients with predialysis fistula after arteriovenous fistula creation / 中华肾脏病杂志

Objective:To explore the influencing factors of hemodialysis (HD) initiation in non-diabetic kidney disease (NDKD) patients with predialysis arteriovenous fistula (AVF) creation.Methods:This was a single-center prospective cohort study. The NDKD patients undergoing predialysis AVF creation were enrolled at the First Affiliated Hospital of Xi'an Jiaotong University from 2015 to 2018. According to the estimated glomerular filtration rate (eGFR, the Chronic Kidney Disease Epidemiology Collaboration equation) and age, patients were divided into different subgroups, eGFR: group 1 [eGFR<10 ml·min -1·(1.73 m 2) -1], group 2 [ eGFR between 10 to 15 ml·min -1·(1.73 m 2) -1], and group 3 [eGFR > 15 ml·min -1·(1.73 m 2) -1]; age: age ≥65 years group and age <65 years group. The primary outcome was defined as the initiation of HD within 1 year after AVF surgery. The second outcome was the use of AVF access at the time of HD initiation. Cox proportional hazard regression was performed to identify which demographic and clinical factors were associated with the initiation of HD after AVF surgery. Logistic regression analysis was performed to investigate factors associated with AVF use at the initiation of HD. Results:A total of 220 patients were enrolled, with age of (48.1±16.2) years, of which 143(65.0%) were males. Overall, the clinical parameters of eGFR, cystatin C, serum albumin, 24h-Urine protein, serum phosphorus were as follows respectively, 7.7 (6.6,9.2) ml·min -1·(1.73 m 2) -1, (3.93±1.12) mg/L, (36.0±4.0) g/L, (2.22±1.36) g, (1.71±0.53) mmol/L. The proportion of patients initiating HD within 6 months ( Fisher=6.832, P=0.020) and the level of hemoglobin ( F=3.112, P=0.047) were higher in group 3 compared to the other two eGFR groups. While the median time interval between AVF creation and HD initiation ( H=6.295, P=0.043) was shorter in group 1. In age <65 years group, the level of serum albumin ( t=2.076, P=0.039), triglyceride ( t=1.995, P=0.048) were higher compared with age ≥65 years group; interestingly, the proportion of patients initiated HD within 3 months ( χ2=4.033, P=0.045) and 6 months ( χ2=5.012, P=0.025) were lower in age <65 years group. The median time interval between AVF creation and HD initiation among these patients was 84 (49,174) days. The patients initiating HD within 3 months, 6 months, and 1 year after AVF creation were 112 (50.9%), 152 (69.1%), and 202 (91.8%), respectively. Multivariate Cox regression analysis indicated that higher cystatin C level ( HR=1.283, 95% CI 1.121-1.469, P<0.001) was associated with earlier HD initiation within 1 year of AVF surgery in NDKD patients. AVF usage was accomplished in 64.3% of patients who initiated HD within 90 days, the ratio was 100.0% in those initiated HD between 91 to 180 days, and 88.0% in those ≥181 days after AVF surgery. No factor was independently associated with AVF use at HD initiation identified by multivariate logistic regression analyses in patients with NDKD. Conclusion:Serum cystatin C level is associated with HD initiation within 1 year of the predialysis AVF creation in NDKD patients.

Effect of Xiaoyu Xiezhuo Drink on Podocyte Apoptosis and Expression of HIF1α in Diabetic Kidney Disease Mice / 浙江中医药大学学报

[Objective]To observe the effect of Xiaoyu Xiezhuo Drink on podocyte apoptosis and hypoxia inducible factor 1 alpha(HIF1α)expression in db/db diabetic kidney disease(DKD)model mice.[Methods]Six db/m mice were chosen as negative control group,eighteen db/db mice were chosen and divided into DKD model group,low dosage Xiaoyu Xiezhuo Drink group,high dosage Xiaoyu Xiezhuo Drink group,with six mice in each group.After gastric irrigation for twelve weeks,the urine was collected to test the levels of protein,β2-microglobulin(β2-MG),albumin/creatinine ratio(Acr),β2-microglobulin/creatinine ratio(β2-MG/Ucr);blood was collected to test the level of triglyceride(TG),total cholesterol(TC),albumin(Alb),blood urea nitrogen(BUN),serum creatinine(Scr);the kidney tissue was collected to observe the pathological change by light and electron microscope,and to test HIF1α,nephrin mRNA by Real-time polymerase chain reaction(RT-PCR).[Results]Compared with negative control group,the levels of urine protein,β2-MG,Acr,β2-MG/Ucr,serum TG,TC,BUN,Scr were increased(P＜0.01),serum Alb was decreased(P＜0.01);glomerular volume increased,capillary loops lobed,mesangial cells and matrix hyperplasia,interstitial inflammation and fibrosis increased,foot process fusion increased,basement membrane thickened;podocyte apoptosis was increased;expression of HIF1α mRNA was elevated(P＜0.01),and nephrin mRNA was descended in kidney tissue of DKD model group(P＜0.01).Compared with DKD model group,the level of urine protein,β2-MG,Acr,β2-MG/Ucr,serum TG,TC,BUN,Scr were decreased(P＜0.01),serum Alb was incresed(P＜0.01);the pathological changes of the kidney was improved;the apoptosis of podocyte was reduced;the expression of HIF1α mRNA was decreased(P＜0.01),and nephrin mRNA was incresed(P＜0.01)in the kidney tissue of varied dosage Xiaoyu Xiezhuo Drink groups.There was no statistical significance in the level of urine protein,β2-MG,Acr,β2-MG/Ucr,serum TG,TC,BUN,Scr,Alb,podocyte apoptosis,and HIF1α,nephrin mRNA in the kidney tissue between different dosage Xiaoyu Xiezhuo Drink groups(P＞0.05).[Conclusion]Xiaoyu Xiezhuo Drink could improve urinary protein,renal function,renal structure lesion and podocyte apoptosis in DKD mice,which perhaps by regulating the expression of HIF1α.

Association between Yang Deficiency Syndrome and the End-point Events of Diabetic Kidney Disease： A Retrospective Cohort Study / 中医杂志

ObjectiveTo investigate the impact of yang deficiency syndrome on the progression to end-point events of diabetic kidney disease （DKD）. MethodsA retrospective study among patients with stage Ⅳ DKD admitted to Dongzhimen Hospital of Beijing University of Chinese Medicine from September 1st， 2016 to September 30th， 2021 was conducted. Data on the patients' general information， clinical indicators including duration of diabetes， duration of proteinuria， history of smoking and drinking， hemoglobin （HGB）， fasting blood glucose （FBG）， albumin （ALB）， serum creatinine （Scr）， urea nitrogen （BUN）， uric acid （UA）， cholesterol （TC） ， triglycerides （TG）， low-density lipoprotein （LDL）， 24-hour urine protein quantification （24h-UTP） and estimated glomerular filtration rate （eGFR）， and TCM syndromes including symptoms， tongue and pulse， and syndrome scores were collected. The patients were divided into exposure group （yang-deficiency group） and non-exposure group （non-yang-deficiency group）. The general information， clinical indicators and incidence rates of end-point events were compared， and the impact of yang deficiency syndrome on the end-point events of stage Ⅳ DKD was analyzed. Survival analysis was performed using Kaplan-Meier method， and multivariate Cox proportional risk models were used to identify independent predictors of end-point events. ResultsA total of 160 patients with stage Ⅳ DKD were included in the study， including 43 cases of yang deficiency syndrome and 117 cases of non-yang deficiency syndrome. Compared to those in the non-yang deficiency group， the waist circumference， BUN and the incidence of end-point events in the yang deficiency group were significantly higher （P＜0.05 or P＜0.01）. Spearman correlation analysis showed that yang deficiency syndrome was positively correlated with incidence of end-point events of stage Ⅳ DKD （r = 0.167， P = 0.035）. Furthermore， 24h-UTP and BUN levels were also positively correlated with end-point events in stage Ⅳ DKD patients （P＜0.01）， while ALB and HGB levels were negatively correlated （P＜0.01）. Kaplan-Meier survival curves showed that yang deficiency syndrome was associated with an increased risk of end-point events （Log Rank P = 0.011）. Moreover， 24h-UTP levels ≥3500 mg， BUN level ≥8 mmol/L， ALB level ＜30 g and HGB level ＜11 g were all associated with the increase of the risk of end-point events （P＜0.05 or P＜0.01）. Multivariate Cox regression analysis showed that yang deficiency syndrome was an independent risk factor for patients with stage Ⅳ DKD to progress into end-point events （HR = 2.36， 1.32 to 4.21； P = 0.004）， as well as 24h-UTP ≥ 3500 mg， BUN ≥ 8 mmol/L， HGB＜11 g and ALB＜30 g （P＜0.05 or P＜0.01）. ConclusionsFor stage Ⅳ DKD， patients with yang deficiency syndrome are more likely to have end-point events， which is an independent risk factor for the progression into end-point events.

Research Progress on the Mechanisms of Berberine Protecting the Kidney in Diabetic Kidney Disease / 中山大学学报(医学科学版)

Diabetic kidney disease （DKD） is a chronic kidney disease caused by diabetes， influenced by genetic and environmental factors and their interaction. It is the primary cause of chronic kidney disease and end-stage renal disease. Recent studies have found， as a natural isoquinoline alkaloid， berberine （BBR） has hypoglycemic， hypolipidemic， antioxidant， anti-inflammatory and anti-fibrotic properties， thus protects against kidney injury in DKD. The mechanisms of action of BBR may involve improving glucolipid metabolism， reducing oxidative stress， alleviating inflammatory responses， mitigating renal fibrosis， regulating DNA methylation， promoting mitochondrial function and modulating the gut microbiota to enhance gut metabolism and clearance. This article systematically reviews the current status of research on the mechanisms of BBR in the treatment of DKD and provides reference for future clinical application of BBR in the treatment of DKD.

Effects of Taohong siwu decoction modified granules on podocyte epithelial-mesenchymal-transition and renal fibrosis in rats with diabetic kidney disease / 中国药房

OBJECTIVE To investigate the effects of Taohong siwu decoction modified granules on podocyte epithelial- mesenchymal-transition （EMT） and renal fibrosis in diabetic kidney disease （DKD） model rats. METHODS Eight rats were selected as normal group （ordinary feed）； the remaining rats were given a high-glucose and high-fat diet combined with intraperitoneal injection of streptozotocin （35 mg/kg） to induce the DKD model. Model rats were randomly divided into model group， irbesartan group [positive control， 13.5 mg/（kg·d）] and modified Taohong siwu decoction group [6.48 g/（kg·d）]， with 8 rats in each group. All groups were given relevant medicine intragastrically， once a day， for 16 consecutive weeks. Twenty-four- hour urinary total protein （24 h UTP） was detected at the end of the 4th， 8th， 12th and 16th week of administration. After the last medication， the body mass， water intake， food intake， urine output， the levels of fasting blood glucose， serum creatinine （Scr） and blood urea nitrogen （BUN） as well as mRNA and protein expressions of P-cadherin， nephrin， α -smooth muscle actin （α-SMA）， Wilms’ tumor gene 1 （WT1）， transforming growth factor-β1（ TGF-β1） and type Ⅳ collagen （Col-Ⅳ） in renal tissue were determined. The pathological and morphological changes in renal tissue were observed and the thickness of the glomerular basement membrane was determined. RESULTS Compared with the model group， 24 h UTP of rats was significantly decreased in modified Taohong siwu decoction group since the 8th weekend （P＜0.05）； the body weight of rats increased significantly， but the amount of water intake and urine decreased significantly； Scr and BUN level， mRNA expression of α-SMA， mRNA and protein expressions of TGF-β1 and Col-Ⅳ were significantly reduced， while the mRNA expressions of P-cadherin， nephrin and WT1 were increased significantly （P＜0.05）； the protein deposition of α-SMA was reduced， protein depositions of P-cadherin， nephrin and WT1 were increased； the pathological damage and fibrosis of renal tissue were relieved； the thickness of glomerular basement membrane was decreased significantly （P＜0.05）. CONCLUSIONS Taohong siwu decoction modified granules can inhibit the EMT of podocyte in DKD model rats， and alleviate renal pathological damage and podocyte damage， thus protecting renal function， and delaying the process of renal fibrosis.

Effects of Taohong siwu decoction modified granules on podocyte epithelial-mesenchymal-transition and renal fibrosis in rats with diabetic kidney disease / 中国药房

OBJECTIVE To investigate the effects of Taohong siwu decoction modified granules on podocyte epithelial- mesenchymal-transition （EMT） and renal fibrosis in diabetic kidney disease （DKD） model rats. METHODS Eight rats were selected as normal group （ordinary feed）； the remaining rats were given a high-glucose and high-fat diet combined with intraperitoneal injection of streptozotocin （35 mg/kg） to induce the DKD model. Model rats were randomly divided into model group， irbesartan group [positive control， 13.5 mg/（kg·d）] and modified Taohong siwu decoction group [6.48 g/（kg·d）]， with 8 rats in each group. All groups were given relevant medicine intragastrically， once a day， for 16 consecutive weeks. Twenty-four- hour urinary total protein （24 h UTP） was detected at the end of the 4th， 8th， 12th and 16th week of administration. After the last medication， the body mass， water intake， food intake， urine output， the levels of fasting blood glucose， serum creatinine （Scr） and blood urea nitrogen （BUN） as well as mRNA and protein expressions of P-cadherin， nephrin， α -smooth muscle actin （α-SMA）， Wilms’ tumor gene 1 （WT1）， transforming growth factor-β1（ TGF-β1） and type Ⅳ collagen （Col-Ⅳ） in renal tissue were determined. The pathological and morphological changes in renal tissue were observed and the thickness of the glomerular basement membrane was determined. RESULTS Compared with the model group， 24 h UTP of rats was significantly decreased in modified Taohong siwu decoction group since the 8th weekend （P＜0.05）； the body weight of rats increased significantly， but the amount of water intake and urine decreased significantly； Scr and BUN level， mRNA expression of α-SMA， mRNA and protein expressions of TGF-β1 and Col-Ⅳ were significantly reduced， while the mRNA expressions of P-cadherin， nephrin and WT1 were increased significantly （P＜0.05）； the protein deposition of α-SMA was reduced， protein depositions of P-cadherin， nephrin and WT1 were increased； the pathological damage and fibrosis of renal tissue were relieved； the thickness of glomerular basement membrane was decreased significantly （P＜0.05）. CONCLUSIONS Taohong siwu decoction modified granules can inhibit the EMT of podocyte in DKD model rats， and alleviate renal pathological damage and podocyte damage， thus protecting renal function， and delaying the process of renal fibrosis.

Zhenwutang Ameliorates Diabetic Kidney Disease in Mice with Spleen-kidney Yang Deficiency via Nrf2/HO-1/GPX4 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect and mechanism of Zhenwutang on renal oxidative damage in the mouse model of diabetic kidney disease with the syndrome of spleen-kidney Yang deficiency via the nuclear factor erythroid 2-related factor-2 （Nrf2）/heme oxygenase-1 （HO-1）/glutathione peroxidase 4 （GPX4） signaling pathway. MethodTwenty-five 7-week-old SPF-grade male db/m mice and 95 7-week-old SPF-grade male db/db mice were adaptively fed for a week. A blank group was set with the db/m mice without treatment， and the other mice were administrated with Rhei Radix et Rhizoma decoction and hydrocortisone for the modeling of diabetic kidney disease with the syndrome of spleen-kidney Yang deficiency. The modeled mice were randomized into the model， irbesartan （25 mg·kg-1）， and high-， medium-， low-dose （33.8， 16.9， 8.45 g·kg-1） Zhenwutang groups （n=15） and administrated with corresponding drugs for 8 weeks. The survival status of mice was observed， and the traditional Chinese medicine （TCM） syndrome score was recorded. The indicators related to spleen-kidney Yang deficiency， fasting blood glucose （FBG）， and renal function indicators were determined. Hematoxylin-eosin staining was employed to observe the histopathological changes of the renal tissue in each group. Biochemical kits were used to determine the oxidative stress-related indicators in the renal tissue. Real-time polymerase chain reaction and Western blotting were employed to determine the mRNA and protein levels， respectively， of Nrf2， HO-1， glutamate-cysteine ligase catalytic subunit （GCLC）， and GPX4 in the renal tissue of mice in each group. ResultCompared with the blank group， the modeling increased the TCM syndrome score （P<0.05）， elevated the estradiol （E2） and FBG levels （P<0.05）， lowered the testosterone （T）， triiodothyronine （T3）， and tetraiodothyronine （T4） levels （P<0.05）， and weakened the renal function （P<0.05）. In addition， the modeling led to glomerular hypertrophy and glomerular mesangial and basal thickening， decreased the catalase （CAT） activity， total antioxidant capacity （T-AOC）， and glutathione （GSH） content （P<0.05）， increased the malondialdehyde （MDA） content （P<0.05）， and down-regulated the mRNA and protein levels of Nrf2， HO-1， GCLC， and GPX4 in the renal tissue （P<0.05）. Compared with the model group， high and medium doses of Zhenwutang decreased the TCM syndrome score and E2 content （P<0.05）， increased the T， T3， and T4 content （P<0.05）， improved the renal function （P<0.05）， alleviated the pathological changes in the renal tissue， increased CAT， T-AOC， and GSH （P<0.05）， reduced MDA （P<0.05）， and up-regulated the mRNA and protein levels of Nrf2， HO-1， GCLC， and GPX4 in the renal tissue （P<0.05）. ConclusionZhenwutang can improve the general state and renal function and reduce the oxidative damage and pathological changes in the renal tissue of db/db mice with spleen-kidney Yang deficiency by regulating the Nrf2/HO-1/GPX4 signaling pathway.

Danggui Buxuetang Inhibits Mitochondrial Fission and Apoptosis of Podocytes in Rat of Diabetic Kidney Disease via AKAP1/Drp1 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo observe the effects of Danggui Buxuetang on the mitochondrial fission and apoptosis of podocytes in the rat model of diabetic kidney disease （DKD） and to explore the protective effect and mechanism of Danggui Buxuetang on DKD rats. MethodSD rats were randomized into a modeling group （n=65， fed with a high-sugar and high-fat diet） and a normal group （n=10， fed with a normal diet）. After 6 weeks， the modeled rats were injected intraperitoneally with streptozotocin （STZ） for the modeling of type 2 diabetes mellitus （T2DM）. Sixty T2DM rats were randomized into model， irbesartan （0.014 g·kg-1）， and low-， medium-， and high-dose （0.36， 0.72， 1.44 g·kg-1， respectively） Danggui Buxuetang groups and administrated with corresponding drugs by gavage for 16 weeks. The levels of fasting blood glucose （FBG） and 24 h urinary protein （24 h UTP） were determined at the end of the 16th week. The pathological changes of the renal tissue were observed by hematoxylin-eosin and Masson staining. The mitochondrial ultrastructure of rat podocytes was observed by transmission electron microscopy. The level of reactive oxygen species （ROS） in the renal tissue of rats was measured by the fluorescent probe labeling of dihydroethidium （DHE）. The apoptosis of renal cells was detected by terminal-deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL）. The expression levels of Synaptopodin， Podocin， and cleaved caspase-3 in renal podocytes were detected by the immunohistochemical method （IHC）. Western blot was employed to determine the protein levels of A-kinase anchoring protein 1 （AKAP1）， phosphorylated dynamin-related protein 1 （p-Drp1）， mitofusin-2 （Mfn2）， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax）. ResultCompared with the control group， the model group showed elevated levels of FBG and 24 h UTP， mesangial hyperplasia， basement membrane thickening， mitochondrial swelling， mitochondrial crista breakage and disorder， and increased ROS and TUNEL-positive cells. In addition， the model group showcased down-regulated expression of Synaptopodin and Podocin， increased expression of cleaved Caspase-3， up-regulated protein levels of AKAP1 and p-Drp1 ， and down-regulated protein levels of Mfn2 and Bcl-2/Bax （P<0.01）. Compared with the model group， high-dose Danggui Buxuetang lowered the levels of FBG and 24 h UTP， alleviated the pathological injuries of the renal tissue and the mitochondrial injury of podocytes， decreased ROS and TUNEL-positive cells， promoted the expression of Synaptopodin and Podocin， inhibited the expression of cleaved Caspase-3， down-regulated the protein levels of AKAP1 and p-Drp1， and up-regulated the protein levels of Mfn2 and Bcl-2/Bax （P<0.05， P<0.01）. ConclusionDanggui Buxuetang may inhibit mitochondrial fission and apoptosis of podocytes and reduce urine protein by regulating the AKAP1/Drp1 pathway， thereby delaying the progression of DKD.

Screening and identification of core genes for diabetic nephropathy / 安徽医科大学学报

Objective @#To screen the core genes of diabetic kidney disease ( DKD) based on bioinformatics , ex- plore the therapeutic targets of DKD , and discuss its possible regulatory mechanism .@*Methods @#The expression da- ta matrix of glomerular transcriptome in patients with DKD in GEO database (GSE30528 , GSE47183) was extrac- ted , and the differentially expressed genes ( DEGs) were screened by bioinformatics methods to identify the core differential genes , and then gene expression and enrichment analysis (GSEA) were conducted to predict effective targets . @*Results @#By screening and identifying the mRNA expression matrix of DKD , five core genes were screened out. Among them , C1orf21 and NPHS1 were significantly down regulated , and CD48 , COL1A2 , and TGFBI were up regulated . NPHS1 and CD48 were significantly related to immune differences , intercellular communication , and cell surface interaction . Through receiver operating characteristic curve ( ROC) analysis and GSEA analysis and drug target prediction , it might be of great significance for the treatment of DKD .@*Conclusion @#The core genes screened in this study have significant correlation with DKD , which may be used as effective markers for the treat- ment of diabetes . And then , this study provides a theoretical basis for the treatment and identification of DKD .

Effect of Intestinal Microecology on Diabetic Kidney Disease Based on Yin-Yang Theory / 中国实验方剂学杂志

Diabetic kidney disease （DKD） is the main cause of end-stage renal disease. Its high prevalence， mortality rate， and medical cost bring a heavy economic burden to society and families， and DKD has become one of the most important public health problems. Intestinal microecology is the most important and complex micro-ecosystem in the human body， which is involved in important life activities such as material and energy metabolism， immune system regulation， and signal transduction， thereby maintaining the dynamic balance of the human internal environment. The dynamic balance between the intestinal microecology and the body is essentially a Yin-Yang balance. Once this balance is broken， intestinal microbiota imbalance， intestinal mucosal barrier damage， immune dysfunction， and reduction of metabolite short-chain fatty acids （SCFAs） will occur， which play an important role in the progression of DKD. From the perspective of the Yin-Yang theory of traditional Chinese medicine （TCM）， the imbalance of intestinal microecology in DKD is equivalent to the excessive or insufficient constraint of Yin and Yang， or Yin deficiency affecting Yang， or Yang deficiency affecting Yin， or waning and waxing of Yin and Yang. For different pathogenesis changes， "Yin disease treated through Yang"， "treating Yin for Yang"， or "treating Yang for Yin" methods are adopted to regulate intestinal microbiota， inhibit immune inflammation， protect intestinal mucosal barrier， and increase SCFAs through TCM， thereby reconciling Yin and Yang to achieve the condition where "Yin is at peace and Yang is compact". Based on the Yin-Yang theory， this paper intended to interpret the scientific connotation of TCM in the treatment of DKD with intestinal microecology as the target and TCM in the treatment of DKD by regulating intestinal microecology as the breakthrough point to provide a novel insight for the occurrence and development of DKD and the mechanism of TCM.

Clinical Prediction Model for Diabetic Kidney Disease Based on Optical Coherence Tomography Angiography / 中山大学学报(医学科学版)

ObjectiveTo construct and validate a clinical prediction model for diabetic kidney disease （DKD） based on optical coherence tomography angiography （OCTA）. MethodsThis study enrolled 567 diabetes patients. The random forest algorithm as well as logistic regression analysis were applied to construct the prediction model. The model discrimination and clinical usefulness were evaluated by receiver operating characteristic curve （ROC） and decision curve analysis （DCA）， respectively. ResultsThe clinical prediction model for DKD based on OCTA was constructed with area under the curve （AUC） of 0.878 and Brier score of 0.11. ConclusionsThrough multidimensional verification， the clinical prediction nomogram model based on OCTA allowed for early warning and advanced intervention of DKD.