Wenyang Huazhuo Fang exerts transient receptor potential cation channel subfamily C member-dependent nephroprotection in a rat model of doxorubicin-induced nephropathy.

OBJECTIVE: To determine the effect of Wenyang Huazhuo Fang (WHF), a Traditional Chinese Medicine decoction, on renal function in a rat model of doxorubicin-induced nephropathy, and to elucidate the underlying mechanism. METHODS: Sprague-Dawley rats were randomly divided into six groups: control, doxorubicin-nephropathy, and prednisone-treated (6.45 mg·kg-1·d－1) doxorubicin nephropathy groups, as well as high- (7.26 g·kg-1·d－1, medium- (2.42 g·kg-1·d－1, and low-dose (0.81 g·kg-1·d－1 WHF-treated doxorubicin-nephropathy groups. The nephropathy rat model was established by two tail vein injections of doxorubicin, followed by prednisone or WHF treatment for 8 weeks. Body weights were monitored and urinary protein was measured every 2 weeks. After the end of the treatment period, the rats were euthanized. Serum biochemical indicators were determined and renal morphological alterations were assessed using histological staining. The expression of transient receptor potential cation channel subfamily C member 6 (TRPC6), stromal interaction molecule 1 (STIM1), and calcium release-activated calcium channel protein 1 (Orai1) was detected using western blotting, and their mRNA levels were examined using quantitative real-time reverse transcription-polymerase chain reaction. RESULTS: WHF treatment was found to significantly ameliorate weight loss, proteinuria, hypoalbuminemia, and dyslipidemia in doxorubicin-nephropathy rats. The protein and mRNA levels of TRPC6, STIM1, and Orai1 were partially, but significantly suppressed by prednisone or WHF treatment. CONCLUSION: Treatment with WHF significantly ameliorates renal injury in a rat model of doxorubicin-induced nephropathy, which could be at least partially related to repression of the TRPC6 pathway.

Polycyclic Aromatic Hydrocarbons in Urban Soil in the Semi-arid City of Xi'an, Northwest China: Composition, Distribution, Sources, and Relationships with Soil Properties.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous in the environment. This study collected a total of 62 urban soil samples from the typical semi-arid city of Xi'an in Northwest. They were analyzed for the composition, distribution, and sources of PAHs as well as the relationships with soil properties. The sum of 16 individual PAHs (∑16PAHs) ranged from 390.6 to 10,652.8 µg/kg with a mean of 2052.6 µg/kg. The average ∑16PAHs decreased in the order of the third ring road (2321.1 µg/kg) > the first ring road (1893.7 µg/kg) > the second ring road (1610.0 µg/kg), and in the order of industrial areas (3125.6 µg/kg) > traffic areas (2551.6 µg/kg) > educational areas (2414.4 µg/kg) > parks (1649.5 µg/kg) > mixed commercial and traffic areas (1332.8 µg/kg) > residential areas (1031.0 µg/kg). The most abundant PAHs in the urban soil were 3- to 5-ring PAHs. Elevated levels of PAHs were found in industrial and traffic areas from the east and west suburbs and the northwest corner of Xi'an as well as the northeast corner in the urban district of Xi'an. PAHs in the urban soil were mainly related to the combustion of fossil fuel (i.e., coal, gasoline, diesel, and natural gas) and biomass (i.e., grass and wood) (variance contribution 57.2%) as well as the emissions of petroleum and its products (variance contribution 29.9%). Soil texture and magnetic susceptibility were the main factors affecting the concentration of PAHs in urban soil. Meanwhile, this study suggested that the single, rapid, and nondeductive magnetic measurements can be an indicator of soil pollution by PAHs.

Characteristics, sources, and health risk of polycyclic aromatic hydrocarbons in urban surface dust: a case study of the city of Xi'an in Northwest China.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental pollutants. Urban surface dust is an important carrier of PAHs. To investigate the characteristics, sources, and health risk of PAHs in urban surface dust, this study collected urban surface dust samples from Xi'an, the largest city in Northwest China and one of the cities with severe smog occurrences in China. The total concentration of 16 US EPA priority PAHs (∑16PAHs) ranged from 5.0 to 48 mg/kg, with an average of 14 mg/kg. The seven carcinogenic PAHs accounted for 21 to 65 % of the ∑16PAHs. Higher levels of PAHs were found in its industrial, traffic, and mixed commercial and traffic districts. The PAHs were dominated by four-ring PAHs, and the predominant components were Fla, Phe, Chy, and Pyr. Multivariate statistical analyses showed that the PAHs originated mainly from the combustion of fossil fuel as well as coal and wood, and petroleum emission. The toxic equivalency quantities (TEQs) of urban surface dustborne PAHs ranged from 0.25 to 8.3 mg/kg, with a mean of 1.8 mg/kg. The 95 % upper confidence limit of incremental lifetime cancer risk (ILCR) due to human exposure to urban surface dustborne PAHs was 8.2 × 10(-5) for children and 7.3 × 10(-5) for adults.

Artemisinin ameliorated proteinuria in rats with adriamycin-induced nephropathy through regulating nephrin and podocin expressions.

OBJECTIVE: To investigate the effects of artemisinin against proteinuria and glomerular filtration barrier damage in rats with adriamycin-induced nephropathy, and the potential mechanism underpinned the action. METHODS: Forty adriamycin rats were randomly divided into two groups with the ratio of 1 : 3; the small-number group served as control group (n = 10), and the rats in the large-number group were treated with adriamycin to induce nephropathy; then they were further randomly assigned into 3 subgroups: benazepril group (n = 10), artemisinin group (n = 10), and adriamycin group (n = 10). The benazepril group and artemisinin group were treated with benazepril suspl (5.0 mg/kg daily) and artemisinin suspl (150 mg/kg daily) respectively after being modeled; those in the control group and adriamycin group were intragastrically administered an equivalent volume of distilled water every day. The treatment after model establishment lasted for a total of 4 weeks. The 24 h uric protein, blood biochemicals, renal pathological changes, renal ultrastrutural changes, Nephrin and Podocin proteins and gene expressions were measured by Coomassie brilliant blue assay, completely automatic biochemical analyzer, light microscope, electron microscopy, Western blot and reverse transcription polymerase chain reaction, respectively. RESULTS: The rats in adriamycin group showed a significant increase in 24 h uric protein excretion, serum total cholesterol (TC), triglyceride (TG), blood urea nitrogen (BUN), serum creatinine (Scr) and decrease in albumin (Alb) (P < 0.05 or P < 0.01). Compared with adriamycin group, artemisinin could reduce uric protein excretion, decrease the serum TC, TG elevation, increase the serum Alb level, up-regulate the expressions of Nephrin and Podocin (P < 0.05 or P < 0.01), but no statistical significance effects on the levels of BUN, Scr in artemisinin group (P > 0.05). The renal pathological and ultrastrutural observation indicate that artemisinin could attenuate the severity of foot process effacement and fusion in the nephropathic rats. CONCLUSION: Artemisinin might have an effect on the nephropathy in rats caused by adriamycin, which may be at least partly correlated with attenu- ation of the severity of foot process effacement and fusion, up-regulation of the expressions of Nephrin and Podocin in the glomeruli in the rats.

"Qufeng Tongluo" acupuncture prevents the progression of glomerulonephritis by decreasing renal sympathetic nerve activity.

ETHNOPHARMACOLOGICAL RELEVANCE: Although the exact mechanism(s) underlying acupuncture remain unknown, acupuncture and acupuncture-like somatic nerve stimulation have been used to treat different kidney diseases and several complications related to them.The aim of this preliminary study was to assess the effectiveness of acupuncture on glomerulonephritis (GN) according to the theory of "Wind-hided renal collaterals" previously proposed. MATERIAL AND METHODS: We used a New Zealand white rabbit model of cationized bovine serum albumin (cBSA)-induced glomerulonephritis and then administered them metoprolol, irbesartan or acupuncture to evaluate the effectiveness of acupuncture treatment and preliminarily explore its potential mechanism. RESULTS: After immunization, our results showed that compared to the cBSA+MET and cBSA+IRB medication groups, "Qufeng Tongluo" significantly lowered parameters of renal function and improved podocyte injury in the 3rd, 6th and 8th weeks of treatment. Moreover, acupuncture increased the protein expression of phosphorylated ERK1/2. CONCLUSIONS: Our study suggests that a potential mechanism by which acupuncture has an antihypertensive effect and can significantly halt deteriorating renal function due to cBSA GN might be mediated by inhibiting the Erk1/2 MAPK pathway to reduce renal sympathetic nerve activity (RSNA).

[Effect of Qufeng Tongluo Recipe on the expressions of connexin 36 protein and gene in rat mesangial cells].

OBJECTIVE: To determine the effect of Qufeng Tongluo Recipe (QFTLR) on the expressions of connexin 36 (Cx36) protein and gene in rat mesangial cells (MCs) and the proliferation of the MCs. METHODS: Serum samples containing Benazepril (Bena) and QFTLR were prepared in line with serum pharmacology methodology. The MCs cultured in vitro were divided into normal control and Lipopolysaccharide (LPS), Bena and QFTLR treated groups. The expressions of Cx36 protein and gene were detected by laser scanning confocal microscope (LSCM), Western blot, immunohistochemical assay and quantitative real time polymerase chain reaction (QRT-PCR) respectively. RESULTS: Compared with the control, higher level of Cx36 protein expression was found in the MCs than treated with LPS (P < 0.01). Both Bena and QFTLR lowered the level of Cx36 protein expression in the MCs treated with LPS significantly (P < 0.01 or P < 0.05). Similar results were found with the expression of Cx36 mRNA. CONCLUSION: QFTLR inhibits the proliferation of rat MCs, possibly through down-regulating the expressions of Cx36 protein and gene.

Qufeng Tongluo Prescription () inhibits mesangial cell proliferation and promotes apoptosis through regulating cell cycle progression.

OBJECTIVE: To study the effects and possible underlying mechanism of Qufeng Tongluo Prescription (, QFTL) on the regulation of mesangial cells (MCs) proliferation and apoptosis. METHODS: The MCs used in this experiment have undergone five passages induced by lipopolysaccharide (LPS). Changes in the proliferation, apoptosis, cell cycle regulatory proteins and mRNA expression levels of the MCs after administration of Benazepril or QFTL were measured by methyl thiazolyl tetrazolium (MTT) reduction assay, flow cytometry, Western blot and quantitative real-time polymerase chain reaction (qRT-PCR), respectively. RESULTS: The addition of Benazepril or QFTL serum inhibited LPS-induced MC proliferation after treatment for 24, 48 and 72 h, respectively (P<0.05 or P<0.01). Moreover, the inhibitory effect is more significant in the QFTL group at 48 h (P<0.05). Compared with the control group, LPS-induced cell proliferation decreased the number of cells in G1 phase versus cells in S and G2/M phases, while the addition of QFTL and Benazepril serum increased the ratio of cells at G1 phase (P<0.05 or P<0.01) to cells at S phase (P<0.01), implicating the cell cycle inhibition effect exerted by QFTL. LPS decreased the level of MC apoptosis, compared with the control group (P<0.05), while QFTL and Benazepril serum increased the level of MC apoptosis (P<0.01). Moreover, the difference between the QFTL group and the Benazepril group was statistically significant (P<0.01). Compared with the control group, the protein and mRNA expression levels of cylinD1, cyclin dependent kinase 2 (CDK2) and p21 were significantly increased (P<0.05 or P<0.01), p27 was decreased but with no statistical significance (P>0.05); After being treated with QFTL and Benazepril serum, the protein and mRNA expression levels of cylinD1, CDK2, p21 were decreased and p27 increased significantly (P<0.05 or P<0.01); Compared with the Benazepril group, QFTL show better effects on protein and mRNA expression levels of cylinD1, CDK2 (P<0.05 or P<0.01) and p21 protein expression (P<0.05). CONCLUSION: QFTL inhibits MCs proliferation, promotes MCs apoptosis through an underlying mechanism of down-regulating the protein and mRNA expression levels of cylinD1, CDK2, p21 and up-regulation of the expression level of p27.

[Effect of acupuncture on renal function and pathologic changes of kidney in rabbits with nephritis].

OBJECTIVE: To observe the effect of acupuncture on cationized bovine serum albumin (C-BSA) nephritis model in rabbits and to explore its mechanism. METHODS: Fifty rabbits were randomly divided into a blank group, a model group, a metoprolol group, a irbesartan group and an acupuncture group, 10 rabbits in each group. The model was established by ear vein intravenous injection with C-BSA. The positive control groups were treated by intragastric administrated with metoprolol and irbesartan, respectively. The acupuncture group was treated by acupuncture at "Fengmen" (BL 12) and "Shenshu" (BL 23). No interventions were added on the blank group and the model group. The changes of blood pressure (BP), heart rate (HR), plasma norepinephrine (NE), serum creatinine (Scr), blood urea nitrogen (BUN) and 24 hours urine protein (24 h UP) in rabbits at the time points of 3rd, 6th and 8th week of treatment were observed. RESULTS: After the model was established, the Scr of (194.30 +/- 20.09) micromol/L, BUN of (9.19 +/- 0.66) mmol/L and 24 h UP of (277.70 +/- 20.09) mg/24 h in the model group were all higher than the Scr of (66.03 +/- 4. 76) micromol/L, BUN of (4.11 +/- 0.71) mmol/L and 24 h UP of (14.28 +/- 1. 47) mg/24 h in the blank group (all P < 0.01), and the diffused mesenteria hyperplasia and the increase of intercapillary cells in the model group were showed in the pathological sections. After 3 weeks of treatment. The Scr of (99.82 +/- 9.29) micromol/L, BUN of (6.32 +/- 0.75) mmol/L and 24 h UP of (189.67 +/- 15.45) mg/ 24 h in the acupuncture group were all decreased significantly, furthermore, the decrease of BP, HR, NE were better than the other treatment groups (P < 0.05, P < 0.01). Except the level of 24 h up and HR at 8th week, other results were as same as the 3rd week. CONCLUSION: Acupuncture can improve the function of kidney, decrease the content of 24 h UP and the underlying therapeutic mechanism could be correlated with that acupuncture can lower excitability of sympathetic nerve and alleviate the renal pathological lesion induced by nephritis.

[Effects of Qufengtongluo Recipe on expressions of cell cycle regulatory proteins in rat mesangial cells].

OBJECTIVE: To observe the effects of Qufengtongluo Recipe (QFTLR) on the expressions of cell cycle regulatory proteins in rat mesangial cells (MCs) in vitro and investigate the mechanism by which QFTLR inhibits MC proliferation. METHODS: Using the methods of serum pharmacology, we studied the expressions of cell cycle regulatory proteins in rat MCs treated with QFTLR by laser scanning confocal microscope and immunohistochemistry. RESULTS: Compared with the normal control cells, the cells challenged with lipopolysaccharide (LPS) showed significantly enhanced expressions of cyclin D1, CDK2, and P21 (P<0.01) and obviously lowered protein expression of P27 (P<0.01). Treatment of the LPS-challenged cells with QFTLR and benazepril both resulted in significantly attenuated expressions of cyclin D1, CDK2, and P21 and obvious increase of P27 expression (P<0.05 or P<0.01), but QFTLR produced stronger effects than benazepril in regulating of cyclinD1, P21 and P27 protein expressions (P<0.05 or P<0.01). CONCLUSION: QFTLR inhibits rat MC proliferation in vitro possibly by down-regulating the cellular expressions of cyclin D1, CDK2, and P21 and up-regulating the expression of P27 protein.

Effect of artemisinin combined with glucocorticoid on the expressions of glucocorticoid receptor α mRNA, glucocorticoid receptor ß mRNA and P300/CBP protein in lupus nephritis mice.

OBJECTIVE: To investigate the therapeutic effects and mechanisms of using artemisinin (Art) combined with glucocorticoid (GC) to treat lupus nephritis (LN) mice. METHODS: Forty hybrid female mice were randomly and equally divided into four groups with the method of random number table: control group, model group, prednisone group administrated with 6.45 mg/(kg·d) prednisone suspension, and Art+prednisone group administrated with 150 mg/(kg·d) Art suspension and 3.225 mg/(kg·d) prednisone suspension. A mice model of LN was established by injection with living lymph cell suspension. The changes of urine protein/24h, the expressions of GC receptor α (GRα) mRNA, GC receptor ß (GRß) mRNA in peripheral blood mononuclear cells (PBMCs), and transcriptional coactivator P300/CBP protein in renal tissue were measured. RESULTS: Compared with the model group, the treatment groups had significant decrease in urine protein/24 h, and renal pathological lesion (P<0.01). In the same groups, the expression of transcriptional coactivator P300/CBP protein in renal tissue and GRα mRNA were significantly increased, and GRß mRNA expression was significantly decreased (P<0.01). And the Art+prednisone group has a better therapeutic effect than the prednisone group (P<0.01). CONCLUSIONS: Art has therapeutic sensitization effects on GC in the LN mice. The underlying mechanism could be correlated with the effect of Art on the increase of the expressions of GRα mRNA and transcriptional coactivator P300 300/CBP protein in renal tissue and on the decrease of the expression of GRß mRNA in PBMC.

[Effects of artemisinin on proliferation, apoptosis and Caspase-3 expression of rat mesangial cell].

OBJECTIVE: To study the effects of artemisinin on proliferation, apoptosis and Caspase-3 active of rat mesangial cell. METHODS: Rat mesangial cells were incubated with different concentrations of artemisinin, the proliferation, apoptosis and Caspase-3 active of rat mesangial cell were measured by MTT assay, fluorescent inverted microscope and enzyme-labeled analytical instruments respectively. RESULTS: Compared with control group, the proliferation and Caspase-3 expression of mesangial cell of three other groups were significantly different (P < 0.01). Compared with dexamethasone group, there were significant difference effects of proliferation and Caspase-3 expression of mesangial cell in other two groups of identical concentration drugs (P < 0. 01), especially in the artemisinin + glucocorticoid (ArtGC) group, and the effects of three different drugs on the mesangial cell Caspase-3 expression, proliferation and apoptosis had the tendency of depend on dosage, and mass mortality of mesangial cell in the mediate-dosage and high-dosage ArtGC group. CONCLUSION: Artemisinin could inhibit the proliferation of mesangial cell, enhance the expression of Caspase-3 and promote the apoptosis in a dose-dependent manner.