Effect of C21 steroidal glycoside of Cynanchum auriculatum on liver and kidney fibrosis through TLR4 pathway / 中国中药杂志

The liver and kidney fibrosis model was established by thioacetamide(TAA) and unilateral ureteral obstruction(UUO) in SD rats. The rats were randomly divided into three groups: model group, low and high-dose groups of C21 steroidal glycosides of Cynanchum auriculatum. Another blank control group was set. Four weeks later, serum was taken to detect the biochemical indexes of liver and kidney function. Urine protein and urine creatinine were detected by kits. Liver and kidney tissue samples were stained with HE and Masson staining, and hydroxyproline content was detected. Western blot was used to detect expressions of fibrotic proteins, inflammatory factors and TLR4 signaling pathways, so as to observe the preventive and therapeutic effects of C21 steroidal glycosides from C. auriculatum on hepatic and renal fibrosis and explore its molecular mechanism. Four weeks later, serum biochemical results showed that liver and kidney functions were seriously damaged, and pathological sections showed that inflammatory cell infiltration, decrease of parenchymal cells, and increase of interstitial fibrosis in liver and kidney tissues. The results showed that low and high doses(150, 300 mg·kg~(-1)) of C21 steroidal glycosides could significantly reduce the collagen deposition and the pathological changes of liver and kidney fibrosis compared with the model group. At the same time, we found that the expression levels of TLR4 and MyD88 signaling pathway proteins were significantly increased in the liver and kidney tissues of the model group, and a large number of NF-κB signaling pathway proteins migrated into the nucleus. On the contrary, the expression levels of TLR4, MyD88 signaling pathway proteins and the nuclear migration of NF-κB were significantly inhibited in the low and high dose groups of C21 steroidal glycosides from C. auriculatum. Therefore, it was speculated that the mechanism of C21 steroidal glycoside for preventive and therapeutic effect on hepatic and renal fibrosis was related to inhibit TLR4/MYD88/NF-κB inflammatory pathway, thus preventing hepatic and renal fibrosis.

Yacon Extract Attenuated Kidney Fibrosis in 5/6-subtotal Nephrectomy Mouse Model by Upregulating HGF and BMP-7 mRNA Expression

@#Introduction: Chronic kidney disease (CKD) leads to tubular injury, kidney fibrosis and anemia. These conditions are influenced by fibrotic and anti-fibrotic substances, such as Transforming Growth Factor beta-1 (TGF-β1), Hepatic Growth Factor (HGF), and Bone Morphogenic Protein-7 (BMP-7). Yacon is an herbal medicine which has not been elucidated in CKD. This study aimed to investigate the effect of ethanolic extract of Yacon leaves on attenuating renal injury in CKD model in mice. Methods: We performed 5/6 subtotal nephrectomy (SN) in male Swiss-Webster mice (3 months old, 30–40 grams) to induce chronic kidney disease, then the mice were sacrificed at day 14. The mice (n=25) were divided into five groups: one SN group, three groups of SN with administration of Yacon extract, and one group of sham operation (SO, with supplementation of 0.1% aquadest). There were three different doses of ethanolic extract of Yacon leaves: 98 mg/kg BW (SN+YK1), 49 mg/kg BW (SN+YK2), and 24.5 BW mg/kg (SN+YK3). Tubular injury, perivascular and interstitial fibrosis were quantified based on histopathological examination. Reverse-transcriptase PCR (RT-PCR) was performed to quantify HGF and BMP-7. Results: SN group demonstrated CKD with elevation of creatinine level, anemia, tubular injury, glomerulosclerosis, and fibrosis. Yacon extract treatment showed attenuation of injury with lower creatinine level, tubular injury, glomerulosclerosis and fibrosis compared to the SN group. HGF and BMP-7 mRNA expressions were higher in Yacon-treated groups than the SN group. Conclusion: Yacon treatment might ameliorate CKD through reducing fibrosis and increasing expression of anti-fibrotic genes.

Effects of 1,25(Oh)2d3 on microrna-130b and transforming growth factor beta 1 in renal tissue of rat models of diabetic nephropathy / 中国组织工程研究

BACKGROUND: 1,25(OH)2D3 plays an important regulatory role in the development of diabetic nephropathy. OBJECTIVE: To explore the effect of 1,25(OH)2D3 on the expression of microRNA-130b and transforming growth factor pi in kidney of diabetic nephropathy rats. METHODS: The study was approved by the Laboratory Animal Ethical Committee of Laboratory Animal Center of Xinjiang Medical University. Twenty-five clean Sprague-Dawley rats were randomly divided into normal control group, diabetic nephropathy+1,25(OH)2D3 group and diabetic nephropathy+peanut oil group. The latter two groups were given calcitriol (1,25(OH)2D3, active vitamin D3, 0.03 jg/kg

Upregulation of p16, Bax and Bcl-2 mRNA Expression Associated with Epithelial Apoptosis and Myofibroblast Proliferation in Kidney Fibrosis Model in Mice

@#Background: Cellular senescence may play a role in the development of kidney fibrosis, but its specific association with apoptosis or proliferation have yet to be determined. Objectives: This study aims to determine the effects of unilateral ureteral obstruction (UUO) on proliferation, cellular senescence and apoptosis in kidney fibrosis. Methods: A unilateral ureteral obstruction (UUO) procedure was performed to induce kidney fibrosis in 24 Swiss mice (3 months old, 30 g–40 g). Mice were sacrificed on day 3 (UUO3, n = 6), day 7 (UUO7, n = 6) and day 14 (UUO14, n = 6). Sham operation (SO) procedures were performed on the control group. The expression of Bcl-2, p16 and Bax mRNA was quantified with reverse transcription polymerase chain reaction (RT-PCR). Immunohistochemical (IHC) staining with anti-Bcl-2 and p53 antibodies was used to determine the localisation of proliferation and apoptosis. Data were analysed using one-way ANOVA followed by a post hoc least significant difference (LSD) test (P < 0.05) Results: RT-PCR analysis showed higher mRNA expression of Bcl-2, p16 and Bax in the UUO groups compared with SO group (P < 0.05). Immunostaining showed that Bcl-2 and p53 expression in tubular epithelium in the UUO groups, except Bcl-2 expression was found in interstitial areas of UUO14 group. Conclusion: Senescence in UUO might be associated with epithelial apoptosis and myofibroblast proliferation.

Effect of Quyu Chencuo Formula () on Renal Fibrosis in Obstructive Nephropathy Rats / 中国结合医学杂志

OBJECTIVE@#To observe the effect of Quyu Chencuo Formula (, QCF) on renal fibrosis in rats with obstructive nephropathy.@*METHODS@#Twenty-four rats were randomly divided into three groups, 4 for sham operation as the control group, 10 for unilateral ureteral obstruction (UUO) model group, and the rest 10 for QCF treating UUO model group. All rats were sacrificed under 3% pentobarbital (50 mg/kg) anesthesia on the 14th day after surgery, then the right kidney samples of rats were harvested for hematoxylin eosin (HE) staining and Masson staining to observe the renal pathological changes. Immunohistochemistry and Western blotting were used to examine the expression of transforming growth factor β1 (TGF-β1), and real-time polymerase chain reaction (RT-PCR) was employed to examine the expressions of TGF-β1, α-smooth muscle actin (α-SMA) and E-cadherin mRNA.@*RESULTS@#HE and Masson staining showed that the renal interstitial of the rats in the control group had no significant fibrotic lesion; in the model group, there were obvious interstitial fibrosis; for the QCF group, there were epithelial cell necrosis, infiltration of lymphocytes and mononuclear cells, aggravated interstitial fibrosis in varied degrees, but the pathological changes were less in the QCF group than in the model group. The immunohistochemistry and Western blotting results showed that the TGF-β1 expression was increased significantly in the model group, while decreased significantly in the QCF group (P<0.05); RT-PCR showed that the mRNA expression of α-SMA and TGF-β1 increased significantly in the model group, while both were significantly decreased in the QCF group compared with the model group (P<0.05). The mRNA expression of E-cadherin was decreased significantly in the model group, and it was significantly increased in the QCF group as compared with the model group (P<0.05).@*CONCLUSION@#QCF may improve renal fibrosis by regulating the expressions of TGF-β1, α-SMA and E-cadherin, and prevent the progress of kidney fibrosis.

Active maintenance of endothelial cells prevents kidney fibrosis

BACKGROUND: Soluble epoxide hydrolase (sEH) expressed by endothelial cells catalyzes the metabolism of epoxyeicosatrienoic acids (EETs), which are vasoactive agents. METHODS: We used a unilateral ureteral obstruction mouse model of kidney fibrosis to determine whether inhibition of sEH activity reduces fibrosis, the final common pathway for chronic kidney disease. RESULTS: sEH activity was inhibited by continuous release of the inhibitor 12-(3-adamantan-1-ylureido)-dodecanoic acid (AUDA) for 1 or 2 weeks. Treatment with AUDA significantly ameliorated tubulointerstitial fibrosis by reducing fibroblast mobilization and enhancing endothelial cell activity. In an in vitro model of endothelial-to-mesenchymal transition (EndMT) using human vascular endothelial cells (HUVECs), AUDA prevented the morphologic changes associated with EndMT and reduced expression of fibroblast-specific protein 1. Furthermore, HUVECs activated by AUDA prevented the epithelial-to-mesenchymal transition (EMT) of tubular epithelial cells in a co-culture system. CONCLUSION: Our findings suggest that regulation of sEH is a potential target for therapies aimed at delaying the progression of kidney fibrosis by inhibiting EndMT and EMT.

Active maintenance of endothelial cells prevents kidney fibrosis

BACKGROUND: Soluble epoxide hydrolase (sEH) expressed by endothelial cells catalyzes the metabolism of epoxyeicosatrienoic acids (EETs), which are vasoactive agents. METHODS: We used a unilateral ureteral obstruction mouse model of kidney fibrosis to determine whether inhibition of sEH activity reduces fibrosis, the final common pathway for chronic kidney disease. RESULTS: sEH activity was inhibited by continuous release of the inhibitor 12-(3-adamantan-1-ylureido)-dodecanoic acid (AUDA) for 1 or 2 weeks. Treatment with AUDA significantly ameliorated tubulointerstitial fibrosis by reducing fibroblast mobilization and enhancing endothelial cell activity. In an in vitro model of endothelial-to-mesenchymal transition (EndMT) using human vascular endothelial cells (HUVECs), AUDA prevented the morphologic changes associated with EndMT and reduced expression of fibroblast-specific protein 1. Furthermore, HUVECs activated by AUDA prevented the epithelial-to-mesenchymal transition (EMT) of tubular epithelial cells in a co-culture system. CONCLUSION: Our findings suggest that regulation of sEH is a potential target for therapies aimed at delaying the progression of kidney fibrosis by inhibiting EndMT and EMT.

Advances of epigenetics in the progression of renal fibrogenesis / 国际儿科学杂志

Epigenetics, which is called second genetic code precisely and controls the total metabolic processes of eukaryotic cells,has an inseparable correlation with human health and diseases. Among all the gene silencing mechanisms of epigenetic modifications,CpG islands of DNA promoter hypermethylation/histone mod-ifications are two major causes. Promoter methylation of specific genes,such as Ras protein activator analogue 1 may lead to aberrant activation of downstream signaling pathways,following by fibroblasts proliferation and kid-ney fibrosis. In recent years,epigenetic modifications have been proved to play a major role in preventing fibro-blasts to return to their quiescent stage,ultimately contributing to fibrosis in the kidney. This paper provides a glimpse of recent studies on epigenetic mechanisms of chronic kidney diseases,especially renal fibrosis,with an aim to provide new insights into pathogenesis research and clinical diagnosis and treatment.

The mechanism of melamine-induced renal damage in rats / 中华泌尿外科杂志

Objective To investigate the mechanism of melamine-induced renal damage in rats.Methods 48 male SD rats were randomly divided into 4 groups with 12 in each group and feed for 3 months.Group A were the control group,feed with standard granule feedstuff and drinking tap water.Group B were stone-induced group,feed with granule feedstuff containing 3％ Mel and drinking tap water.Group C were feed with granule feedstuff containing 3％ Mel and drinking water containing 2％ taurine.Group D were feed with standard granule feedstuff and drinking water containing 2％ taurine.Every week 24 h urine was collected to test PH,SCr,uric acid,protein,8-IP,H2O2 and Mel level.All rats were sacrificed at the end of 3 months.Blood creatinine detection,renal pathology analysis ( HE and Oil ep-red O dyeing,immunohistochemical) and mitochondria separation and detection were undertaken. ResultsMel was not detedted in urine of Group A and Group D.The urine concentration of Mel in Group B and Group C in 1 week,2 weeks,3 weeks,4 weeks were 3.16 ±0.45,4.39 ±0.213,5.40 ±0.28,5.50 ±3.26 and 3.52 ±0.49,4.32 ± 0.135,5.34 ± 0.40,5.46 ± 2.99 mg/ml,respectively.Compared with Group A,the Mel concentration in urine of Group B and C were drug exposure time dependent.In Group A,the urine protein,urine creatinine clearance,serum creatinine,and renal/weight ratio were 6.45 ± 1.45 mg/24 h,28.0 ± 7.4mmol/l,0.56 ±0.03 ml · min-1 · 100g-1,2.29 ±0.89 mg/g,while in Group B and C,the urinary protein urine,serum creatinine,creatinine clearance,kidney/weight ratio were 14.56 ± 7.69,56.8 ± 5.2,0.29 ±0.05,4.16 ±0.27 and 16.44 ±6.29,55.8 ±7.4,0.30 ±0.07,4.40 ±0.56,respectively.Compared with group A,in Group B and C,the urinary protein increased significantly,urine creatinine clearance reduced,serum creatinine reduced,and renal/weight ratio increased.Compared with Group B,the improvement of renal function in Group C was not significant,and the decrease of serum creatinine and urinary protein were not obvious (P ＞ 0.05).In Group B and C,the urine H2O2,8-IP and mitochondrial oxidatie detection reagent SOD,GSH-PX numerical were 28.5 ± 5.2 mmol/1,3.26 ± 1.6 pg/ml,21.1 ± 7.8 U/mg prot,19.0 ±2.5 energy unit and 26.7 ±4.8 mmol/l,2.99 ±8.5 pg/ml,20.3 ±6.9 U/mg prot,17.9 ±4.8 energy unit,respectively.The difference between Group B and C was not statistically significant (P ＞0.05).Pathological analysis showed Mel was mainly concentrated in crystal tubular lumen (Group B and C),kidney interstitial damage was apparent,and kidney inflammation and fibrosis progressive developed with the increase of the drug exposure time. Conclusions Mel can induce kidney damage and stone formation in rats,and stone was mainly in tubular location in inner medullary zone.It is not the oxidative stress way that Mel leads to kidney damage.

Influence of tranilast on the cyclosporine A-induced epithelial-to-mesenchymal transition in human renal tubular epithelial cells / 中华器官移植杂志

Objective To study the effect of tranilast on cyclosporine A (CsA)-induced epithelial-to-mesenchymal transition in human renal tubular epithelial cells, and investigate the mechanism of its antifibrotic effect. Methods Cultured HK-2 cells were divided into four groups: (1)In the control group, cells were treated without any medicine; (2) The cell were treated with CsA (4. 2μmol/L) for 72 h; (3) The cells were treated with a combination of CsA (4. 2 μmol/L) and tranilast (100μmol/L); (4) The cells were treated with tranilast (100 μmol/L) alone for 72 h.Morphological changes of the cells were assessed by phase-contrast microscopy. The immunofluorescence and Western blotting were adopted to detect the expression of E-cadherin, α-SMA and OPN mRNA and proteins respectively. Results Tranilast could markedly ameliorate the morphological changes of HK-2 cells stimulated by CsA. The irmmunofluorescence staining revealed the expression of E-cadherin was markedly decreased in HK-2 cells stimulated with CsA for 72 as compared with the control group, while the expression of α-SMA and OPN was significantly higher in CsA group than the control group. The expression of E-cadherin in the CsA + Tranilast group was higher than the CsA group, while the expression of α-SMA and OPN in the CsA + Tranilast group was lower than the CsA group. Western blotting showed that protein expression level of E-cadherin in CsA group was dramatically lower than that in the control group (P＜0. 05), while that of α-SMA and OPN in CsA group was significantly higher than in the control group (P＜0.05). The protein expression level of E-cadherin in HK-2 cells in the CsA + Tranilast group was markedly higher than in the CsA group (P＜0.05), and that of α-SMA and OPN in CsA + Tranilast group was significantly lower than in the CsA group (P＜0. 05). Conclusion Tranilast can block the CsA-induced epithelialto-mesenchymal transition in HK-2 cells probably by suppressing the expression of OPN.

Prossion CHEN Zhi-qiang approaching the present-day application of Biejiajian Pills / 中华中医药杂志

Biejiajian Pills was created by ZHANG Zhong-jing of Han Dynasty. Its feature was harmonizing qi-blood, appllying drugs with nature of cold and heat, and simultaneous application of purging-tonifying therapy, it was a distinguished prescription of treating malarial nodule. Today, the prescription was applied in treating liver fibrosis and had obvious effect. According to the TCM theory, Professor CHEN Zhi-qiang identified the theory of treating kidney by Biejiajian Pills on basis of analyzing the etiopathogenisis and pathogenesis of liver fibrosis and kidney fibrosis. It has made certain progress in the treatment kidney fibrosis by ancient medical prescription.

Effect of Niaoduqing Capsule on Kidney Fibrosis Parameters in Chronic Renal Failure / 广州中医药大学学报

ObjectiveTo observe the therapeutic effect of Niaoduqing Capsule(NC),which has the actions of lifting clear Qi and purging turbidity,for chronic renal failure(CRF) and to investigate its influence on kidney fibrosis parameters.【Methods】A prospective study was carried out in 78 CRF patients.The patients were equally randomized into two groups.Besides the basic treatment of feeding with small-dose high-quality protein,controlling blood pressure,maintaining water-electrolyte and acid-base balance,Group A received oxyamyli tectus aldehydum and group B received NC.Four weeks constituted one treatment course and the treatment lasted 2 courses.Before and after treatment,the scores of main symptoms and syndrome were counted,and laboratory parameters such as hematoglobin(Hb),red blood cells(RBC) count,serum creatinine(SCr),blood urea nitrogen(BUN),creatinine clearance(CCr),albumin(ALB),total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),serum laminin(LN),precollagen-Ⅲ(PC-Ⅲ) and hyaluronic acid(HA) were assayed.【Results】Total scores of main symptoms and scores of most single symptom were lower,and effect on the symptoms and total therapeutic effect were better in group B than those in group A(P0.05).【Conclusion】NC has an effect on relieving symptoms of CRF,and improving renal function and kidney fibrosis parameters,indicating that NC exerts a better counteraction on kidney fibrosis.