Objective To investigate the roles and significances of MMP-2 and MMP-9 in diffuse proliferative lupus nephritis by repeated renal biopsy.Methods Seventeen patients diagnosed by renal biopsy as WHO typeⅣlupus nephritis were analyzed by immunohistochemistry staining for MMP-2 and MMP-9. Double staining for MMP-2 and MT1-MMP,MMP-9 and CD68 were also performed.Patients had repeated renal biopsy after followed up for 2.5 years.The relationship between expressions of gelatinases and pathological activity index and clinical data were studied.Results MMP-2 immunoreactivity was detected in normal controls and was increased in diffuse proliferative lupus nephritis.MMP-9 staining,which was almost negative in normal giomeruli,was increased much more significantly in diffuse proliferative lupus nephritis. The immunoreactivity of MMP-2 and MMP-9 was positive in MT1-MMP staining and CD68-positive macrophages, respectively.The expression of MMP-2 and MMP-9 was reduced by 70% and 62% in 10 patients whose clinical condition was partially alleviated,while the expressions in 7 patients whose clinical condition was not alleviated,were only reduced by 27% and 32%.The staining for MMP-2 and MMP-9 were correlated with activity index of lupus nephritis and proteinuria.Conclusion Up-regulation of gelatinases expression in diffuse proliferate lupus nephritis is correlated to activity index of the disease.

Objective To explore the role of tissue inhibitor of metalloproteinase-1(TIMP-1) during renal senescence by using human TIMP-1 transgenic mice.Methods Renal histological changes of wild type mice and transgenic mice at the age of 3,12,24 months were observed by periodic acid-schiff(PAS)staining of paraffin sections.The numbers of F4/80 positive cells were detected by immunofluoreseence.The protein expressions of TIMP-1,TIMP-2,matrix metalloproteinase(MMP)-9,MMP-2,intercellular adhesion molecule-1(ICAM-1),transforming growth factor?1(TGF-?1),collagenⅢand collagenⅣwere detected by Western blot.The activities of gelatinases and TIMP-1 were examined by gelatin zymography and reverse zymography respectively.Results Focal renal fibrosis was found in two genotypes with aging.At the age of 24 months,compared with wild type,in kidneys of transgenic type,the expressions and activities of gelatinases were dowregulated (MMP-2:2.08?0.20 vs.3.39?0.43;MMP-9:4.02?0.82 vs.6.72?1.40,all P＜0.05);the expressions of collagenⅢ,collagenⅣ,ICAM-1,and TGF-?1 were upragulated(0.72+0.11 vs.0.57?0.09;0.84?0.13 vs.0.6?0.11,0.72?0.12 vs.0.53?0.07; 0.69?0.12 vs.0.45?0.09,all P＜0.05),and the numbers of F4/80 positive cells were increased (18.8?4.4 vs.12.7?3.6,P＜0.05)with the upregulated expression and activity of TIMP-1(1.10?0.18 vs.0.62?0.09;50.75?7.25 vs.20.64?3.50,P＜0.05).Conclusions TIMP-1 could promote age-related renal fibrosis through enhancing inflammation reaction by ICAM-1 upregulation.

OBJECTIVETo observe the effect of compound shenhua tablet (CST) on the residual kidney expressed macrophage migration inhibition factor (MIF) in rats.

METHODSCST was used to treat 5/6 nephrectomized rats for 12 weeks and the conditions of blood pressure, urinary protein, blood biochemical indices (creatinine, blood urea nitrogen), kidney pathologic change and MIF expression were observed.

RESULTSCST could significantly lower the serum levels of creatinine (P < 0.05), and 24 hrs urinary protein (P < 0.01), reduce the MIF expression and macrophage infiltration in renal glomerulus and tubular mesenchym, and lower the degree of renal glomerular sclerosis and interstitial fibrosis.

CONCLUSIONThe inhibition on the highly expressed MIF may be an important mechanism of the drug in restraining chronic inflammation in residual kidney, delaying the sclerosis and fibrosis progression and protecting renal function.

Albuminuria ; blood ; Animals ; Creatinine ; blood ; Drugs, Chinese Herbal ; pharmacology ; Fibrosis ; pathology ; Kidney ; metabolism ; pathology ; Macrophage Migration-Inhibitory Factors ; metabolism ; Male ; Nephrectomy ; Rats ; Rats, Wistar ; Tablets

BACKGROUNDTissue inhibitor of matrix metalloproteinase-1 (TIMP-1) is related to the aging of many organs, but few data are available on the change of TIMP-1 in liver aging. The purpose of this study was to investigate the expression and role of TIMP-1, matrix metalloproteinase-2 (MMP-2) and MMP-9 in the process of natural aging in the livers of normal and transgenic mice, and to detect the effects of TIMP-1 on oxidative level and anti-oxidative ability of the livers of transgenic young mice.

METHODSNormal and transgenic mice were divided into 3 groups according to their age: 3-month-old group (n = 5), 12-month-old group (n = 5) and 24-month-old group (n = 5). Histopathological changes of the liver were observed after HE and Masson staining. The messenger RNA (mRNA) levels of TIMP-1, MMP-2 and MMP-9 were determined by semi-quantitative reverse transcriptional polymerase chain reaction; protein expression was measured by Western blot in the livers of normal and transgenic mice of various ages. Changes in levels of superoxide dismutase (SOD), monoamine oxidase (MAO), malondialdehyde (MDA) as well as oxidative and anti-oxidative ability were measured.

RESULTSHistologically, more fatty degeneration and collagen deposition were found in the aging livers of transgenic mice than in those of the normal mice as their age of months increased. The mRNA and protein expressions of TIMP-1 were significantly high in the oldest animals. The histopathological changes, mRNA and protein expressions of TIMP-1 increased significantly in the liver of transgenic mice as compared with normal mice. The expression of MMP-2 and MMP-9 showed a minor change in the process of aging. Liver change and collagen deposition were not observed in young mice, but the activity of SOD decreased (P < 0.05), and the activity of MAO (P < 0.01) and the content of MDA increased in the liver of transgenic mice (P < 0.01).

CONCLUSIONSThe expression of TIMP-1 is significantly high in the liver of transgenic mouse in the process of aging, indicating that the oxidative level increases and the anti-oxidative ability decreases in the liver of transgenic mouse. TIMP-1 plays an important role in the process of liver aging.

Aging ; metabolism ; Animals ; Female ; Liver ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 2 ; analysis ; genetics ; Matrix Metalloproteinase 9 ; analysis ; genetics ; Mice ; Mice, Transgenic ; Monoamine Oxidase ; analysis ; RNA, Messenger ; analysis ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; analysis ; genetics

OBJECTIVETo investigate the reno protective effect of Shenhua recipe on the experimental model of 5/6 renal ablation.

METHOD5/6 renal ablation rats were underlying this experiment. They were administered Shenhua, irbesartan respectively by gavage during 12 weeks. Body weight, systolic blood pressure, proteinuria, Scr, BUN, total protein, albumin, Glycero and cholesterol were measured. Histologic glomenular and tubulointerstitial damage scores were measured at 12 weeks.

RESULTThe treated groups showed significantly less histologic glomerular and tubulointerstitial damage scores at 12 weeks. The plasma albumin were higher ( P < 0.05), urine protein excretion rates, serum cholesterol and creatinine were lower than in nontreated group, but arterial blood pressure was not significantly different in the three Shenhua treated groups compared with nontreated group.

CONCLUSIONShenhua can retard the progression of chronic renal injury in the 5/6 renal ablation without changes in systolic blood pressure.

Albumins ; metabolism ; Animals ; Astragalus membranaceus ; chemistry ; Atractylodes ; chemistry ; Blood Pressure ; drug effects ; Cholesterol ; blood ; Creatinine ; blood ; Curcuma ; chemistry ; Disease Progression ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Kidney Failure, Chronic ; etiology ; pathology ; prevention & control ; Kidney Glomerulus ; pathology ; Male ; Nephrectomy ; adverse effects ; methods ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar

The aim of this study was to investigate the immunological function regulated by Fufang Hongjingtian capsule (HJT) in mice. The mice were given ig HJT 25, 250 and 750 mg/kg, once daily, for 30 - 38 d, respectively. The mice in control group were given ig corresponding solvent. After the last time of administration, the immunological parameters of the mice were measured. The results showed that compared with negative control group, the delayed type hypersensitivity, spleen lymphocyte proliferation and number of spleen IgM antibody forming cells increased in HJT groups. In conclusion the HJT has the effect to improve the immunological functions of mice.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Immunoglobulin M ; immunology ; Lymphocyte Count ; Lymphocytes ; cytology ; Mice ; Mice, Inbred Strains ; Rhodiola ; Spleen ; cytology ; immunology

OBJECTIVETo observe the effect of Compound Shenhua Tablet (, SHT) on the sodium-potassium- exchanging adenosinetriphosphatase (Na(+)-K(+)-ATPase) in the renal tubular epithelial cells of rats with acute ischemic reperfusion and to investigate the mechanisms underlying the effects of SHT on renal ischemic reperfusion injury (RIRI).

METHODSFifty male Wistar rats were randomly divided into the sham surgery group, model group, astragaloside group [150 mg/(kg·d)], SHT low-dose group [1.5 g/(kg·d)] and SHT high-dose group [3.0 g/(kg·d)], with 10 rats in each group. After 1 week of continuous intragastric drug administration, surgery was performed to establish the model. At either 24 or 72 h after the surgery, 5 rats in each group were sacrificed, blood biochemistry, renal pathology, immunoblot and immunohistochemical examinations were performed, and double immunofluorescence staining was observed under a laser confocal microscope.

RESULTSCompared with the sham surgery group, the serum creatinine (SCr) and blood urea nitrogen (BUN) levels were significantly increased, Na(+)-K(+)-ATPase protein level was decreased, and kidney injury molecule-1 (KIM-1) protein level was increased in the model group after the surgery (P<0.01 or P<0.05). Compared with the model group, the SCr, BUN, pathological scores, Na(+)-K(+)-ATPase, and the KIM-1 protein level of the three treatment groups were significantly improved at 72 h after the surgery (P<0.05 or P<0.01). And the SCr, BUN of the SHT low- and high-dose groups, and the pathological scores of the SHT high-dose group were significantly lower than those of the astragaloside group (P<0.05). The localizations of Na(+)-K(+)-ATPase and megalin of the model group were disrupted, with the distribution areas overlapping with each other and alternately arranged. The severity of the disruption was slightly milder in three treatment groups compared with that of the model group. The results of immunofluorescence staining showed that the SHT high-dose group had a superior effect as compared with the astragaloside group and the SHT low-dose group.

CONCLUSIONSThe SHT effectively alleviated RIRI caused by ischemic reperfusion, promoted the recovery of the polarity of renal tubular epithelial cells, and protected the renal tubules. The therapeutic effects of SHT were superior to those of astragaloside as a single agent.

Acute Disease ; Animals ; Blood Urea Nitrogen ; Cell Adhesion Molecules ; metabolism ; Chromatography, Liquid ; Creatinine ; blood ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Fluorescent Antibody Technique ; Immunoblotting ; Kidney Function Tests ; Kidney Tubules ; blood supply ; drug effects ; enzymology ; pathology ; Low Density Lipoprotein Receptor-Related Protein-2 ; metabolism ; Male ; Rats ; Rats, Wistar ; Reperfusion Injury ; drug therapy ; enzymology ; pathology ; Saponins ; analysis ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Staining and Labeling ; Tablets

BACKGROUNDHuman urate anion exchanger (hURAT1) as a major urate transporter expressed on renal tubular epithelial cells regulates blood urate level by reabsorbing uric acid. Antibody is an important tool to study hURAT1. This study aimed, by genetic immunization, to produce mouse anti-hURAT1 polyclonal antibody with high throughput and high specificity and to detect the location of hURAT1 in human kidney.

METHODSHuman renal total RNA was isolated and the entire cDNA of hURAT1 was amplified by RT-PCR. The sequence of intracellular high antigenicity fragment (A280 to R349) was chosen by prediction software of protein antigenicity, and its cDNA was amplified from cDNA of hURAT1, and then cloned into pBQAP-TT vector to construct recombinant plasmid pBQAP-TT-hURAT1-210 for genetic immunization. Mice were inoculated with this recombinant plasmid and two other adjuvant plasmids, pCMVi-GMCSF and pCMVi-Flt3L, which helped to enhance the antibody's generation. After four weeks, the mice were sacrificed to obtain the anti-hURAT1 antibody from serum. The antibody was identified by western blot analysis and immunohistochemistry. At the same time, rabbit anti-hURAT1 antibody was produced by protein immunization. The specificity and efficiency between the rabbit and mouse anti-hURAT1 antibody were compared by western blot analysis and immunohistochemistry.

RESULTSThe entire cDNA of hURAT1 and cDNA of its intracellular high immunogenic fragment were amplified successfully. Recombinant plasmid pBQAP-TT-hURAT1-210 for genetic immunization was confirmed by restriction digestion and sequencing. Both the mouse anti-hURAT1 antibody and rabbit anti-hURAT1 antibody recognized 58 kD hURAT1 and 64 kD glycosylated hURAT1 protein bands in western blot. Immunohistochemically, hURAT1 was located at the brush border membrane of renal proximal tubular cells. In addition, the throughput and specificity of the mouse anti-hURAT1 antibody were higher than those of the rabbit anti-hURAT1 antibody.

CONCLUSIONGenetic immunization can generate anti-hURAT1 polyclonal antibody of high throughput and specificity.

Animals ; Antibodies ; analysis ; Blotting, Western ; Carrier Proteins ; analysis ; immunology ; Female ; Humans ; Immunization ; Immunohistochemistry ; Kidney ; chemistry ; Male ; Mice ; Organic Anion Transporters ; analysis ; immunology ; Organic Cation Transport Proteins ; Plasmids ; Rabbits

OBJECTIVETo investigate the impact of a traditional Chinese medicinal compound known as Fufang Shenhua Tablet (SHP) on the expression of Toll-like receptors (TLRs) during renal ischemia-reperfusion injury (IRI)-induced acute kidney injury (AKI) in rats.

METHODSA total of 28 Wistar rats were randomly divided into five groups: (1) pseudo-operation control group, (2) ischemia-reperfusion model group, (3) Astragaloside group, (4) high-dose SHP group, and (5) low-dose SHP group. There were four rats in the pseudo-operation group and six rats in each of the other groups. The accepted ischemia-reperfusion model was established after a 7-day gavage intervention, and pathological changes and renal function were observed, using an enzyme-linked immunosorbent assay (ELISA) to detect interleukin 8 (IL-8) and interferon gamma (IFN-γ) levels, as well as immunohistochemical staining to detect altered levels of TLR2 and TLR4 expression in renal tissue.

RESULTSAfter 24 h, renal pathological damage and the expression levels of serum creatinine (Scr), IL-8, IFN-γ, TLR2, and TLR4 were significantly higher in the model group as compared with the pseudo-operation group (P<0.05). In addition, at 24 h the above indicators decreased significantly in the Astragaloside group, high-dose SHP group and low-dose SHP group as compared with the ischemia-reperfusion model group (P< 0.05). TLR2 and TLR4 expression levels were significantly reduced in the SHP treatment and Astragaloside group as compared with the pseudo-operation group (P<0.05). Further, the high-dose SHP group showed significantly less renal damage score and decreased levels of TLR expression than those of low-dose SHP group and Astragaloside group (all P<0.05).

CONCLUSIONSHP can alleviate the renal structural and functional damage caused by IRI-induced AKI in rats by reducing the damage of renal pathology, which may reduce inflammatory cytokine levels by downregulating the expression of TLRs in renal tissue in a dose-dependent manner.

Acute Kidney Injury ; etiology ; metabolism ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Immunohistochemistry ; Interferon-gamma ; blood ; Interleukin-8 ; blood ; Kidney Tubules ; drug effects ; metabolism ; Male ; Rats ; Rats, Wistar ; Reperfusion Injury ; complications ; Tablets ; Toll-Like Receptor 2 ; metabolism ; Toll-Like Receptor 4 ; metabolism

OBJECTIVETo investigate liver and kidney lesions in HBV-GN patients and relationship between them and provide evidences to make early diagnosis of HBV-GN.

METHODSReviewing the clinicopathological and laboratory indexes of 205 patients with HBV-GN diagnosed by renal biopsy in our hospital from September 1995 to November 2008.

RESULTSHBV-GN account for 5.6% of all renal biopsies at the same time. Among them, 157 (76.5%) patients were male,123 (60%) was 19-45 years-old. 95 (46%) patients break out with kidney disease. HBsAg, HBeAg, HBcAg were the most common HBV makers. 102 (49.8%) patients present nephrotic syndrome, 18 (8.8%) suffered kidney dysfunction; 18 patients with hepatic cirrhosis. Patients with or without liver disfunction got no different in clinic manifestation and renal pathology. With the rising of the content of HBV-DNA in surum, the urinary protein increases. Renal data shows that membranous nephropathy(MN) was the most frequent type (60.5%).

CONCLUSIONThe peak incidence of HBV-GN is in the twentieth to forth decade of life. There was a 3:1 predominance of males. Nephrotic syndrome was the most common clinic manifestation and membranous nephropathy was the most common pathology. 10% persent patisnts had loss of renal function at the time of renal biopsy. The HBV copies in serum correlated with the albuminuria. HBV-GN patients had desynchroneity lesions in kidney and liver. As the high rate of HBV infection in China, It needs to prevent the kidney damage in HBV infectious people and to elevate early diagnosis and therapy.

Adult ; DNA, Viral ; blood ; Female ; Glomerulonephritis ; pathology ; virology ; Hepatitis B ; pathology ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Male ; Middle Aged ; Young Adult