OBJECTIVETo study the effects of core proteoglycan on the transdifferentiation of human renal tubular epithelial cell induced by transforming growth factor beta1 (TGF-beta1) in vitro.

METHODThe cultured HK-2 cells were divided into six groups: A. negative control group; B. 10 ng/ml TGF-beta1 group; C. 10 ng/ml core proteoglycan treated group; D. 100 ng/ml core proteoglycan treated group; E. 10 ng/ml TGF-beta1 + 10 ng/ml core proteoglycan group; F. 10 ng/ml TGF-beta1 + 100 ng/ml core proteoglycan group. The changes in configuration of HK-2 cells were inspected 48 hours after adding the stimulating factor. At the same time, changes in mRNA of keratin, alpha-smooth muscle actin, vimentin were analyzed.

RESULTSCompared with group A, group B showed great changes in the morphology of cells, most cells converted into spindle shape, like fibroblast; groups E and F, especially group F showed significantly reduced spindle shape cells. Compared with group A, groups C and D had no significant changes in morphology of cells Compared with 10 ng/ml TGF-beta1 group and negative control, the mRNA expression of alpha-smooth muscle actin and vimentin had significant increase, but that of keratin reduced (P < 0.05). However, after combined treatment with TGF-beta1 and core proteoglycan, alpha-smooth muscle actin and vimentin expression were reduced significantly, while expression of keratin was up-regulated. Single core proteoglycan treated group and negative control group had no dramatic differences (P > 0.05).

CONCLUSIONTGF-beta1 can induce the transdifferentiation of human renal tubular epithelial cell and core proteoglycan has some inhibitory effect on transdifferentiation of human renal tubular epithelial cell induced by TGF-beta1 in vitro.

Actins ; physiology ; Bone Morphogenetic Protein 7 ; metabolism ; Cadherins ; metabolism ; Cell Differentiation ; drug effects ; physiology ; Cell Line ; Cell Shape ; Cell Transdifferentiation ; Cells, Cultured ; Connective Tissue Growth Factor ; pharmacology ; Epithelial Cells ; cytology ; drug effects ; physiology ; Fibroblasts ; drug effects ; pathology ; Humans ; Kidney ; pathology ; Kidney Tubules ; pathology ; Kidney Tubules, Proximal ; pathology ; Proteoglycans ; chemistry ; pharmacology ; Transforming Growth Factor beta ; genetics ; pharmacology ; Transforming Growth Factor beta1 ; chemistry ; pharmacology ; Vimentin ; metabolism

OBJECTIVETo evaluate the toxicity of Radix Aristolochiae and Radix Inulae, and to supply the toxicity experimental data that Radix Inulae supersedes Radix Aristolochiae in clinic.

METHODA long dose of Radix Aristolochice and Radix Inulae was given intragastrically to rats for six months, then drug withdrawal for a month. The hematology and biochemical indicators were measured, and the pathologic changes of kidney, liver, stomach and urinary bladder were examined.

RESULTThe rats of Radix Aristolochice showed serious toxic responses of renal tubule atrophy and necrosis, meanwhile, the levels of BUN, Cr and NAG were increased obviously. Hepatonecrosis, renal tubular necrosis, gastric carcinoma and bladder carcinoma were discovered with pathologic assaying. But the rats of Radix Inulae did not.

CONCLUSIONRadix Aristolochiae could damage kidney and liver, and cause gastric carcinoma and bladder carcinoma by intensive toxicity. Radix Inulae could take the place of Radix Aristolochiae to use in clinic.

Acetylglucosaminidase ; urine ; Animals ; Aristolochia ; chemistry ; Blood Urea Nitrogen ; Creatinine ; blood ; Drugs, Chinese Herbal ; isolation & purification ; toxicity ; Female ; Inula ; chemistry ; Kidney Tubules ; drug effects ; pathology ; Liver ; drug effects ; pathology ; Male ; Necrosis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Stomach ; drug effects ; pathology ; Stomach Neoplasms ; chemically induced ; Urinary Bladder ; drug effects ; pathology ; Urinary Bladder Neoplasms ; chemically induced

OBJECTIVETo study the effect of Astragalus mongholicus (AM) on the expression of transforming growth factor-beta1 (TGF-beta1) in SD rats with unilateral ureteral occlusion (UUO) and to elucidate the mechanisms underlying the renoprotective effects of AM.

METHODFifty-four Sprague-Dawley rats were randomly divided into 4 groups: sham-operation group, the UUO group and AM treatment group. After administration of AM (10 g kg(-1) d(-1)) for 3, 7 and 14 days, the dynamic histological changes of renal interstitial tissues were observed and renal damage including tubular impairment and interstitial fibrosis were quantified on HE and Masson stained tissue sections. The expression of TGF-beta1 and alpha-smooth muscle actin (alpha-SMA) was measured by immunohistochemistry staining sections. The mRNA of TGF-beta1 and alpha-SMA were reverse transcribed and quantified by real-time PCR. The expression of TGF-beta1 protein were assessed by Western blot.

RESULTRenal damage was exacerbated and the expression of alpha-SMA and TGF-beta1 were all significantly increased in UUO group compared with those of sham-operation group (P<0.05) at each time point. Tubular impairment and interstitial fibrosis were alleviated, and up-regulations of expressions of TGF-beta1 and alpha-SMA were significantly suppressed by AM treatment (P<0.05).

CONCLUSIONAM can ameliorate renal interstitial fibrosis induced by UUO in vivo. The mechanisms of its antifibrotic effects might be related with the down-regulation of TGF-beta1 expression and suppression of tubular epithelial myofibroblast transdifferentiation in the progress of renal interstitial fibrosis.

Actins ; genetics ; Animals ; Astragalus Plant ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation ; drug effects ; Kidney Tubules ; drug effects ; metabolism ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Transforming Growth Factor beta1 ; genetics ; Ureteral Obstruction ; metabolism ; pathology ; prevention & control

OBJECTIVETo study whether total saponins of Panax notoginseng (PNS) can prevent renal interstitial fibrosis occurrence through blocking the IL-1alpha induced tubular epithelial cell and decrease the secretion of extracellular matrix (ECM).

METHODSNormal rats' tubular epithelial cells NRK52E were cultured in vitro, their morphological changes were observed by inverted phase contrast microscope and scanning electron microscope. Flow cytometry technique and immuno-histochemical method was used to detect the expression of alpha-smooth muscle actin (alpha-SMA), and ELISA was used to quantitatively detect the fibronectin (FN) in the supernatant

RESULTSIL-1alpha could induce the transdifferentiation of tubular epithelial myofibroblast, showing hypertrophy of cells elongated and fusiform-shaped, with significantly enhanced expression of alpha-SMA and increased secretion of FN (P<0.05). After adding PNS of different concentrations, the morphology of cells restored close to normal tubular epithelial cells, with the increase of alpha-SMA expression and FN secretion significantly inhibited (P<0.05) in dose-dependent manner (P<0.05). But addition of different dosages of PNS alone showed no effect on tubular cells.

CONCLUSIONIL-1alpha could induce the transdifferentiation of tubular epithelial myofibroblast, promote the deposition of ECM component FN. PNS could inhibit IL-1alpha induced the transdifferentiation of NRK52E and secretion of ECM, therefore, PNS could be taken as a new drug for prevention and treatment of renal interstitial fibrosis and terminal stage of renal diseases.

Actins ; analysis ; Animals ; Cell Differentiation ; drug effects ; Cell Line ; Epithelial Cells ; cytology ; Fibronectins ; analysis ; Fibrosis ; prevention & control ; Interleukin-1 ; pharmacology ; Kidney ; pathology ; Kidney Tubules ; cytology ; Panax ; chemistry ; Rats ; Saponins ; isolation & purification ; pharmacology

OBJECTIVETo investigate the effect of L-arginine (L-arg) on early compensatory renal growth (CRG), tubulointerstitial accumulation of extracellular matrix (ECM), long term survival rate and renal scarring in rats with 5/6 nephrectomy (SNx).

METHODSThe experiment included four groups of rats (n = 5 each group): (1) Sham group, (2) SNx group, (3) SNx + L-arg group, and (4) Sham + L-arg group (L-arg 1% in drinking water). Parameters related with CRG and early tubulointerstitial expression of ECM and alpha smooth muscle actin (alphaSMA) were evaluated by immunohistochemistry at day 30. The survival rate and the extent of renal scarring in the rats were observed at day 120.

RESULTSL-arg significantly increased the early CRG of SNx rats as determined by the wet kidney weight (P < 0.05), total protein (P < 0.01), and DNA content (P < 0.001) of the remnant kidney. The cell proliferation in the tubulointerstitium as determined by immunostaining for proliferative cell nuclear antigen (PCNA) also markedly increased (P < 0.05). Furthermore, glomerular volume (Vg) in the SNx rats treated by L-arg increased 16% compared with that in the SNx group (P = 0.06). More specifically, L-arg significantly increased the interstitial immunostaining for collagen III, IV, and fibronectin (P < 0.05, 0.01, respectively), which were positively correlated with the increased expression of alpha-SMA (P < 0.001, respectively). In the long term experiment, L-arg significantly reduced the survival rate (P < 0.05) and increased the severity of renal scarring in the SNx rats (P < 0.05).

CONCLUSIONThis preliminary study suggested that early supplementation with L-arg might exacerbate the progression of renal scarring in rat remnant kidney.

Actins ; metabolism ; Animals ; Arginine ; pharmacology ; Cicatrix ; etiology ; mortality ; pathology ; Disease Progression ; Extracellular Matrix ; metabolism ; Fibrosis ; Immunohistochemistry ; Kidney ; drug effects ; pathology ; surgery ; Kidney Tubules ; chemistry ; drug effects ; pathology ; Male ; Muscle, Smooth ; chemistry ; Nephrectomy ; adverse effects ; Rats ; Rats, Wistar ; Severity of Illness Index ; Survival Rate

OBJECTIVETo observe the renal function, histology and the levels of interleukin-1beta (IL-1beta), interleukin-6 (IL-6) in rats administered with Aristolochi manshriensis (AM) and Longdan Xiegan decoction (LDXG) containing AM.

METHODDivided SD rats into control, AM and LDXG groups (n = 10 per group) at random and administrated orally for 8 weeks with distilled water, AM (6 g x kg(-1) x d(-1)) and LDXG (17. 5 g x kg(-1) x d(-1), containing AM 6 g x kg(-1) x d(-1)), respectively. Measured renal functional parameters in serum, urine and IL-1beta, IL-6 in renal homogenate, and observe renal histologic lesions at the end of 8 weeks.

RESULTTwo kinds of water abstract induced significantly renal injury in rats, IL-1beta, IL-6 levels in renal homogenate were significantly increased compared with controls, histological section showed inflammatory cell infiltration play important role in renal tubulointerstitium damage. LDXG caused less seriously damage than AM.

CONCLUSIONAM and LDXG in experimental dosage can induce renal injury, inflammatory cell infiltration in renal tubulointerstitium and high levels of IL-1beta, IL-6 presumpt that the nephrotoxicity of AM and LDXG refered to the immune mechanism. LDXG has light nephrotoxicity due to its restraining the inflammatory reaction caused by AM.

Administration, Oral ; Animals ; Aristolochia ; chemistry ; Blood Urea Nitrogen ; Creatinine ; blood ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; toxicity ; Interleukin-1beta ; metabolism ; Interleukin-6 ; metabolism ; Kidney ; drug effects ; metabolism ; pathology ; Kidney Tubules ; drug effects ; metabolism ; pathology ; Male ; Plants, Medicinal ; chemistry ; Proteinuria ; blood ; chemically induced ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the renal injury in rats induced by Longdan Xiegan decoction (LDXGD) containing different dosages Aristolochia manshriensis.

METHODSD rats were divided into four groups at random, and were fed with three kinds of LDXGD 13, 14.5, 17.5 g x kg(-1) (containing respectively A. manshriensis 1.5, 3, 6 g x kg(-1)) and distilled water respectively for 12 weeks. Renal functional parameters on 4,8,12 w were determined and changes of histomorphology in rats on the end of experiment were observed.

RESULTThe LDXGD containing low dose (1.5 g x kg(-1)) A. manshriensis did't induce significantly renal injury in rats during 12 weeks; the LDXGD containing midst dose(3 g x kg(-1)) A. manshriensis induced light damage of proximal convoluted tubule epithelial cells in rats during 12 weeks; the LDXGD containing high dose(6 g x kg(-1)) A. manshriensis induced significantly renal injury in rats after administed 4 weeks. Along with the lasting of administration, the degree of injury became more seriously. The main renal injury location was in proximal convoluted tubule.

CONCLUSIONThe renal toxicity of LDXGD is correlated with the dose of A. manshriensis and the time of administration. The LDXGD containing low dose A. manshriensis has relative security. However, the LDXGD containing high dose A. manshriensis can induce renal injury.

Acetylglucosaminidase ; urine ; Animals ; Aristolochia ; chemistry ; toxicity ; Blood Urea Nitrogen ; Creatinine ; blood ; Dose-Response Relationship, Drug ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; toxicity ; Kidney Diseases ; chemically induced ; metabolism ; pathology ; Kidney Tubules, Proximal ; drug effects ; pathology ; Male ; Plants, Medicinal ; chemistry ; toxicity ; Proteinuria ; chemically induced ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; beta 2-Microglobulin ; urine ; gamma-Glutamyltransferase ; urine

OBJECTIVEFollowing the former report, we continue to observe the chronic renal tubular-interstitial injury induced by Radix Aristolochiae Fangchi Extract(RAFE) in rats in order to understand whether RAFE in different doses causes the renal tubular-interstitial injury or not.

METHODRAFE at the dose of 25.0 mg x kg(-1) x d(-1), 120.0 mg kg(-1) x d(-1) and 200.0 mg x kg(-1) x d(-1) and aristolochic acid (AA, 10.0 mg x kg(-1) d(-1)) was interruptedly administrated by gastric tube for 22 w and 4 w durg withdrawal. Blood, urine and kidney were taken out respectively in 17 w, 22 w and 26 w to measure the indexes of renal function. The morphology of kidney was observed, and Masson staining of kidney were made respectively to compare RAFE groups with AA group.

RESULTPathological changes of renal tissue forms were as follows: All RAFE groups and AA group could develop the pathological process of renal tubular injury-chronic renal interstitial fibrosis. The pathologic changes of RAFE were similar with AA.

CONCLUSIONRAFE at all doses administrated interruptedly by gastric tube above 13 w caused chronic renal tubulo-interstitium fibrosis. The renal injury in functions and tissue forms in rats were similar with AA closely. The results showed that AA was the main toxic composition of RAFE.

Animals ; Aristolochia ; chemistry ; toxicity ; Aristolochic Acids ; isolation & purification ; toxicity ; Blood Urea Nitrogen ; Body Weight ; drug effects ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; toxicity ; Female ; Fibrosis ; blood ; chemically induced ; pathology ; Kidney Tubules ; pathology ; Male ; Nephritis, Interstitial ; blood ; chemically induced ; pathology ; Plant Roots ; chemistry ; toxicity ; Plants, Medicinal ; chemistry ; toxicity ; Proteinuria ; chemically induced ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo compare the sensitivity of early renal injury induced by mercury-containing medicine in rats, including urinary N-acetyl-beta-D-glucosdminidase (NAG), beta2-microglobulin (beta2-MG), retinol binding protein (RBP) and clusterin (CLU).

METHODBadu Shengji San(BDSJS), a mercury-containing preparation of traditional Chinese medicine, was adopted as the mercury contact drug. The lowest effective toxic dose was used to observe its effect on serum creatinine (SCr), blood urea nitrogen (BUN), and such early renal injury indicators as NAG, RBP, beta2-MG and CLU and compare the sensitivity of tested indicators.

RESULTCompared to the broken skin group, groups with administration of 60 and 120 mg x kg(-1) doses of BDSJS showed no obvious difference in SCr and BUN when kidney indicators is remarkably increased and obvious pathological changes were found in kidney tubules but with significant increase in the urinary level of CLU and the levels of NAG and RBP. H&E staining of renal tubule showed that exposure of 30 mg x kg(-1) BDSJS had no significant morphological changes, but at the same concentrations, the level of RBP was markedly increased. Urinary beta2-MG levels were markedly decreased in BDSJS 30, 60 mg x kg(-1) group rats, whereas 120 mg x kg(-1) dose group showed no obvious change in urinary beta2-MG levels.

CONCLUSIONUrinary RBP, NAG and CLU were more sensitive than SCr and BUN as indicators for early renal injury in the order of RBP > NAG > CLU, and urinary RBP, NAG would increase earlier than beta2-MG.

Acetylglucosaminidase ; urine ; Animals ; Blood Urea Nitrogen ; Clusterin ; urine ; Creatinine ; blood ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; toxicity ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Kidney Tubules ; drug effects ; metabolism ; pathology ; Male ; Mercury ; blood ; metabolism ; toxicity ; urine ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Retinol-Binding Proteins ; urine ; Skin ; drug effects ; injuries ; Time Factors ; beta 2-Microglobulin ; urine

OBJECTIVETo study the mercury accumulation in injured skin rats induced by Badu Shengji San (BDSJS), a traditional Chinese medicine preparation for external use.

METHODInjured skin rats were treated with BDSJS for consecutively 4 weeks. During the 4 weeks and the following 4 weeks after the drug withdrawal, samples were collected for determining mercury contents in blood, urine and kidney, with urinary N-acetyl-beta-D-glucosaminidase(NAG) and beta2-microglobulin (beta2-MG) as indicators of renal toxicity and serum biochemical indicators of hepatic and renal functions. Additionally, activated partial thromboplastin time (APTT), prothrombin time (PT) and kidney and renal pathological changes were also observed.

RESULTCompared to injured skin rats, mercury contents of blood, urine and kidney were increased significantly in low, middle and high-dose BDSJS groups administered for consecutive 4 weeks. The levels of mercury showed decreases in urine (89%, 78%, 93%) and kidney (55%, 51%, 57%), and blood mercury concentration recovered to the normal range in low, middle and high-dose BDSJS groups after the drug withdrawal for 4 weeks. Kidney coefficient and beta2-MG were remarkably increased and renal tubular epithelial cell swelling could be found in the high-dose group, and kidney coefficient, beta2-MG and renal morphology basically recovered to the normal levels after the drug withdrawal for 4 weeks.

CONCLUSIONThe administration of BDSJS for consecutively 4 weeks can cause mercury accumulation in blood and mainly in kidney. Once the accumulated mercury concentration of kidney reaches a certain level, renal tubular epithelial cells would be injured. 1.1 mg x cm(-2) of BDSJS is proved to be safe and 2.2 mg x cm(-2) can cause mild but reversible injury in the function of kidney which can be recovered after drug withdrawal for 4 weeks.

Acetylglucosaminidase ; urine ; Animals ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; toxicity ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Female ; Kidney Tubules ; drug effects ; metabolism ; pathology ; Male ; Mercury ; blood ; metabolism ; toxicity ; urine ; Rats ; Rats, Sprague-Dawley ; Skin ; drug effects ; injuries ; Time Factors ; beta 2-Microglobulin ; urine