miRNA-93-5p in exosomes derived from M2 macrophages improves lipopolysaccharide-induced podocyte apoptosis by targeting Toll-like receptor 4.

Diabetic nephropathy (DN) is a common complication of diabetes mellitus which can result in renal failure and severely affect public health. Several studies have revealed the important role of podocyte injury in DN progression. Although, the involvement of exosomes derived from M2 macrophages has been reported in podocyte injury, the underlying molecular mechanism of M2 macrophage-secreted exosomes has not been fully elucidated. Our study suggests that M2 macrophages mitigate lipopolysaccharide (LPS)-induced injury of podocytes via exosomes. Moreover, we observed that miR-93-5p expression was markedly upregulated in exosomes from M2 macrophages. Inhibition of miR-93-5p derived from M2 macrophage exosomes resulted in apoptosis of LPS-treated podocytes. Additionally, TLR4 showed the potential to bind to miR-93-5p. Subsequently, we validated that TLR4 is a downstream target of miR-93-5p. Further findings indicated that silencing of TLR4 reversed the renoprotective effects of miR-93-5p-containing M2 macrophage exosomes on LPS-induced podocyte injury. In summary, our study demonstrated that M2 macrophage-secreted exosomes attenuated LPS-induced podocyte apoptosis by regulating the miR-93-5p/TLR4 axis, which provides a new perspective for the treatment of patients with DN.

Kaempferol ameliorates Cisplatin induced nephrotoxicity by modulating oxidative stress, inflammation and apoptosis via ERK and NF-κB pathways.

Anticancer drug like Cisplatin are associated with serious problem like nephrotoxicity. The effect of Kaempferol is a plant-derived flavonoid compound. The present work evaluated the effect of Kaempferol in mouse model of Cisplatin mediated nephrotoxicity also the involved mechanism. Oxidative stress, kidney function, histology, inflammation, apoptosis, level of proteins, Nrf2 translocation and its effect on cascades such as NF-κB and ERK were studied. It was observed that the pre-treatment of KPF reduced the Cisplatin mediated oxidative stress, inflammation, apoptosis and ameliorated renal injury and its functioning. Kaempferol suppressed the Cisplatin induced infiltration of mononuclear cells, levels of TNF-α, iNOS, IL-12, activation of NF-κB, phosphorylation of IκBα and nuclear translocation of p65 in renal tissues. Also KPF attenuated Cisplatin mediated phosphorylation of p38, ERK1/2 and JNK in renal tissues. KPF also corrected the levels of renal antioxidants and elevated the nuclear levels of HO-1 and Nrf2 in renal tissues. KPF attenuated the Cisplatin mediated apoptosis via down-regulating the levels of TP53, Bax/Bcl2 imbalance, activating caspase-3/9 and PARP. The outcomes conclude that KPF ameliorates Cisplatin-mediated nephrotoxicity by modulating oxidative stress, inflammation and apoptosis via ERK and NF-κB pathway.

Association between CYP17 T-34C rs743572 and breast cancer risk.

Association between CYP17 T-34C (rs743572) polymorphism and breast cancer (BC) risk was controversial. In order to derive a more definitive conclusion, we performed this meta-analysis. We searched in the databases of PubMed, EMBASE and Cochrane for eligible publications. Pooled odds ratios (ORs) with 95% confidence intervals (95% CIs) were used to assess the strength of association between CYP17 T-34C polymorphism and breast cancer risk. Forty-nine studies involving 2,7104 cases and 3,4218 control subjects were included in this meta-analysis. In overall, no significant association between CYP17 T-34C polymorphism and breast cancer susceptibility was found among general populations. In the stratified analysis by ethnicity and source, significant associations were still not detected in all genetic models; besides, limiting the analysis to studies with controls in agreement with HWE, we also observed no association between CYP17 T-34C polymorphism and breast cancer risk. For premenopausal women, we didn't detect an association between rs743572 and breast cancer risk; however, among postmenopausal women, we observed that the association was statistically significant under the allele contrast genetic model (OR = 1.10, 95% CI = 1.03-1.17, P = 0.003), but not in other four models. In conclusion, rs743572 may increase breast cancer risk in postmenopausal individuals, but not in premenopausal folks and general populations.

[Effect of Qufeng Tongluo Recipe on expression of desmin and CD2AP proteins in adriamycin-induced nephropathy rats: an experimental research].

OBJECTIVE: To investigate the effect of Qufeng Tongluo Recipe (QTR) on the expression of desmin and CD2-associated protein (CD2AP) in adriamycin-induced nephropathy rats. METHODS: The adriamycin-induced nephropathy rat model was induced by a disposable intravenous injection of adriamycin. The model was successfully established after 3 weeks. Rats were then randomly divided into the blank control group (Group A, n =12), the model control group (Group B, n = 8), the small, medium, large dose QTR group (Group C, n = 8; Group D, n = 8; Group E, n = 8), and the positive control group (Group F, n = 8). From the fourth week normal saline was given to rats in Group A and Group B, QTR 1.0 g/mL, 2.1 g/mL, and 4.2 g/mL was respectively administered to those in Group C, D, and E. Prednisone 25 mg/kg was given to rats in Group F. All medication was performed by gastrogavage at 10 mL/kg, once daily, for 28 successive days. 24-h urinary protein excretion and sera biochemical indices were determined during medication. At the end of the experiment, ultrastructure was observed, mRNA expression of desmin, mRNA and protein of CD2AP were detected by Real-time PCR and Western blot. RESULTS: (1) Compared with Group B, 24-h urinary protein excretion significantly decreased in Group C, D, E, and F (P < 0.05). (2) Compared with Group B, Alb in Group C, D, and E increased (P < 0.05) and TC significantly decreased (P < 0.05). TG significantly increased in Group F (P < 0.05). (3) Results of electron microscope showed, compared with Group B, the morphology of foot cells was improved to various degrees in Groups D, E, and F, especially the foot process structure and the number of foot processes were significantly improved, which was more obviously shown in Group D and Group E. (4) mRNA expression of desmin, mRNA and protein of CD2AP increased in adriamycin-induced nephropathy rats (P < 0.05). After intervention, when compared with Group B, mRNA expression of desmin and CD2AP were significantly lower in Group C, D, E, and F (P < 0.05). (5) Compared with Group A, expression of desmin and CD2AP significantly increased (P < 0.05). Compared with Group B, the expression of desmin protein were obviously lower in Group C, D, E, and F, and the protein expression of desmin obviously decreased in Group D, E, and F (P < 0.05). The protein expression of desmin and CD2AP gradually decreased in Group C, D, and E (P < 0.05). Compared with Group F, the expression of CD2AP protein obviously increased in Group C and D (P < 0.05); the expression of CD2AP protein obviously decreased in Group E (P < 0.05); the expression of desmin protein was higher in Group C, D, and E (P < 0.05). CONCLUSION: QTR's therapeutic effect on adriamycin-induced nephropathy rats might be achieved through altered expression of desmin and CD2AP.

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.

Effects of asiaticoside on levels of podocyte cytoskeletal proteins and renal slit diaphragm proteins in adriamycin-induced rat nephropathy.

AIMS: Centella asiatica has been used to treat kidney diseases in Chinese traditional medicine. Asiaticoside (an extraction of C. asiatica) exerts a variety of pharmacological effects including immunomodulatory and anti-inflammatory functions. However, the mechanism of asiaticoside in the treatment of renal diseases remains largely unknown. This study investigated the molecular mechanism of asiaticoside in treating adriamycin-induced nephropathy of rats. MAIN METHODS: Sixty-two SD male rats were randomly divided into normal control group (n=12) and nephropathy group (n=50). Except for the normal control group, rats were injected with adriamycin (6mg/kg) via the tail vein to induce nephropathy. Adriamycin induced nephropathic rats were divided into untreated group, prednisone group (25mg/kg), and asiaticoside groups with various dosages (8, 16 and 32mg/kg). Samples of urine and serum, tissue of kidney were collected for analysis after treatments for four weeks. Morphological changes were evaluated under light microscope and electron microscope. Synaptopodin, desmin, nephrin and podocin mRNA and protein were determined by RT-PCR and Western blotting. KEY FINDINGS: Compared to the untreated nephropathy group, asiaticoside treatment mitigated histological damages, decreased 24-hour urine protein excretion and total cholesterol, increased serum albumin. Asiaticoside treatment increased the mRNA and protein levels of synaptopodin, nephrin and podocin in a dose-dependent manner. Furthermore, asiaticoside treatment decreased the mRNA and protein levels of desmin. SIGNIFICANCE: Asiaticoside can mitigate adriamycin-induced nephropathy in rats, which is associated with the increase in synaptopodin, nephrin and podocin gene expression, and the decrease in desmin gene expression.

[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 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.

[Effect of artemisinin on the expressions of GRalpha mRNA, GRbeta mRNA and P300/CBP protein in lupus nephritis mice].

OBJECTIVE: To investigate the effects of Artemisinin on the expressions of GRalpha mRNA, GRbeta mRNA and P300/CBP protein in lupus nephritis 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 Artemisinin (Art) suspension. A mice model of LN was established by injection with living lymph cell suspension. The expressions of GC receptor alpha (GRalpha) mRNA, GC receptor beta (GRbeta) mRNA in peripheral blood mononuclear cells (PBMCs), and transcriptional coactivator P300/CBP protein in renal tissue were respectively measured by the technique of RT - PCR and immunohistochemical assay. RESULTS: Compared with the model group. The expression of transcriptional coactivator P300/CBP protein in renal tissue and GRa mRNA in PBMCs of treatment groups was increased significantly, GRbeta mRNA expression was significantly decreased (P < 0.05 or P < 0.01). And the Art group had a better effect on the expression of GRalpha mRNA and transcriptional coactivator P300/CBP protein than that of the prednisone group (P < 0.01). CONCLUSION: The underlying therapeutic mechanism may be correlated with the regulation of Art on the expressions of GRalpha mRNA, GRbeta mRNA in PBMC and transcriptional coactivator P300/CBP protein in renal tissue.

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 Qufengtongluo Recipe on podocin mRNA expression and podocyte morphology in rats with adriamycin-induced nephropathy].

OBJECTIVE: To observe the effects of Qufengtongluo Recipe (QFTLR) on the expression of podocin mRNA and podocyte morphology in rats with adriamycin-induced nephropathy (AN), and explore the possible mechanism mediating the therapeutic effect of QFTLR on nephropathic proteinuria. METHODS: SD rats were randomized into normal control group, AN model group (established by a single injection of adriamycin via the tail vein), and 3 intervention groups with QFTLR, prednisone, or benazepril treatment. After the corresponding treatments, the expression of podocin mRNA in the renal tissues was detected by RT-PCR methods, and the morphological changes of the podocytes were examined by electron microscope. RESULTS: Compared with the normal control group, the AN model group showed significantly lowered expressions of podocin mRNA (P<0.01) with reduced podocytes and widening, fusion or even absence of the foot processes (FP). Intervention with QFTLR significantly increased the expression of podocin mRNA (P<0.01) and the number of podocytes, and obviously lessened the structural changes of the FP. CONCLUSION: QFTLR can produce therapeutic effect against nephropathic proteinuria possibly by up-regulating the expression of podocin mRNA and improving the morphological changes of the podocytes.

[Effects of Qufengtongluo recipe on proteinuria and glomerular filtration membrane in rats with adriamycin-induced nephropathy].

OBJECTIVE: To assess the therapeutic effect of Qufengtongluo (QFTL) recipe against proteinuria and glomerular filtration membrane damage in rats with adriamycin-induced nephropathy (AN). METHODS: Fifty-six SD rats were randomized into normal control (A) and AN model groups. In the AN model group, the rat AN models established by a single intravenous injection of adriamycin via the tail vein were subdivided into model (B), QFTL recipe (C), prednisone (D), and benazepril (E) groups 3 weeks after adriamycin injection. The 24-h urinary protein level was measured and the expression of anionic sites on the filtration membrane was evaluated using electron microscope with PEI staining. Nephrin expression on the glomerular filtration membrane was detected with indirect immunofluorescence assay. RESULTS: Compared with group A, the model group showed significantly increased level of 24-h urinary protein (P<0.01), suggesting successful establishment of the AN model. Treatment with QFTL recipe obviously lowered the 24-h urinary protein (P<0.01), and increased the expression of anionic sites and nephrin on the glomerular filtration membrane in the AN rats (P<0.01). CONCLUSION: QFTL recipe can effectively decrease 24-h urinary protein, improve the symptoms, and up-regulate the expressions of anionic sites and nephrin on the glomerular filtration membrane in rats with AN.

Therapeutic effect of artemisinin on lupus nephritis mice and its mechanisms.

In this study, we investigated the therapeutic effect of artemisinin (Art) on lupus nephritis mice and its mechanisms by comparing the differences between lupus nephritis (LN) mice given Art and control mice in molecular biology, immunohistochemistry, and histopathology. The results showed that Art could remarkably relieve the symptoms, decrease the level of urine protein/24 h, and alleviate pathological renal lesions. The differences among the four groups in the expression of the NF-κBp65 protein, nuclear factor-κB (NF-κB) activity, and the expression of transforming growth factor-ß1 (TGF-ß1) mRNA in renal tissue suggested that Art can lower the serum levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) and inhibit the expression of the NF-κBp65 protein and NF-κB and TGF-ß1 mRNA in the renal tissues of LN mice. These results proved that it is reliable and effective to use Art to treat LN mice, and its therapeutic mechanisms should closely be related to the fact that Art can obviously decrease the serum levels of TNF-α and IL-6 and down-regulate the expression of the NF-κBp65 protein and NF-κB and TGF-ß1 mRNA in renal tissues.

[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.

[Effects of qufengtongluo recipe on expressions of HSPG mRNA and protein in adriamycin-induced nephropathy rats].

OBJECTIVE: To investigate the expression of HSPG in glomerular base membrane of adriamycin-induced nephropathy (AN) rats, and the effect of Qufengtongluo recipe on HSPG mRNA expression and proteinuria in AN rats. METHODS: One hundred forty rats were used in this study, including 32 rats in normal control group. AN was induced in the left rats by a single tail intravenous injection of adriamycin. Three weeks later, 90 AN rats were randomly divided into five groups; the nephropathy group (B, n=18), the Qufeng group (C, n=18), Qufeng and prednisone group (D, n=18), prednisone group(E,n=18) and benazepri group (F, n= 18). The rats in these five groups were treated with different combination of Qufeng recipe and prednisone. In each group, renal tissue samples were collected at week 3 and 7. The distribution, expression of HSPG was examined by indirect immunofluorescence, and semi-quantity RT-PCR, respectively. RESULTS: (1) In AN rats, the diffuse fusion and effacement of foot processes were observed when model established. (2) Compared with nephropathy group, the average fluorescence intensity of HSPG dramatically increased in Qufeng group and prednisone group (P < 0.01), similarly, it also increased in D and F groups (P < 0.01). (3) Compared with nephropathy group, the expression of HSPG mRNA was significantly up-regulated in other groups. (P < 0.01), especially in C and F groups. There was significant negative correlation between the expression of HSPG and quantity of 24-hour proteinuria. CONCLUSION: The abnormal expression of HSPG and their altered distributions may be an important molecular mechanism that leads to the occurrence and development of proteinuria in AN rats. The effect of Qufengtongluo recipe on nephrotic syndrome might be related to the alteration of HSPG expression and distribution in glomerulus.

[Effect of Qufengtongluo recipe on expression of nephrin mRNA in adriamycin-induced nephropathy in rats].

OBJECTIVE: To investigate the expression of nephrin mRNA in adriamycin-induced nephropathy (AN) in rats, and to explore the effect of Qufengtongluo recipe on proteinuria in AN rats and on the expression of nephrin mRNA. METHODS: Adriamycin nephropathy was induced by a single tail intravenous injection of adriamycin. We randomly divided 140 rats into a normal control group (n=32) and a nephropathy model group (n=108). Three weeks later, 90 AN rats were randomly divided into 5 groups: a model group, a qufeng group, a qufeng and prednisone group, a prednisone group, and a benazepri group (18 rats in each group). They were treated respectively, and renal tissue samples were collected at week 0, 3, 5, and 7, respectively. The distribution and expression of nephrin mRNA were examined by indirect immunofluorescence and semi-quantity RT-PCR. RESULTS: In the AN rats, the diffuse fusion and effacement of foot processes were observed at week 3. The fluorescence intensity of nephrin and the expression of nephrin mRNA significantly increased in the qufeng group and the prednisone group compared with the model group at the week 7 (P<0.01). CONCLUSION: Abnormal expression of nephrin is the important molecular mechanism that leads to the occurrence and development of proteinuria in AN rats. Qufengtongluo recipe has effect on nephrotic syndrome through altering the expression and distribution of nephrin in glomerulus.

[Qufeng Tongluo decoction inhibits lipopolysaccharide-induced rat mesangial cell proliferation in vitro by suppressing NF-kappaB activation and reducing TGF-beta1 and IL-6 mRNA expressions].

OBJECTIVE: To investigate the effect of Qufeng Tongluo (QFTL) decoction on lipopolysaccharide (LPS)-induced rat mesangial cell proliferation and explore the possible mechanisms underlying the therapeutic effects. METHODS: Thirty-two rats were randomized into normal control, glomerulonephritis model, QFTL treatment and positive control groups, and serum samples were obtained from these groups. Rat mesangial cells with or without LPS exposure were treated with the sera, and the expression of nuclear factor-kappaB (NF-kappaB ) was detected using electrophoretic mobility shift assay (EMSA), and the expressions of transforming growth factor-beta1 (TGF-beta1) and interleukin-6 (IL-6) mRNAs were detected with RT-PCR. RESULTS: QFTL decoction inhibited the activation of NF-kappaB in LPS-stimulated rat mesangial cells stimulated by LSP, and lowered the expressions of TGF-beta1 and IL-6 mRNA. CONCLUSION: QFTL decoction can inhibit LPS-induced rat mesangial cell proliferation by decreasing the expression of TGF-beta1 and IL-6 mRNA as a result of suppression NF-kappaB activation.

[Effects of Qufeng Tongluo Recipe on proliferation of and TGF-beta1 and IL-6 mRNA expressions in glomerular mesangial cells from rats].

OBJECTIVE: To investigate the effects of Qufeng Tongluo Recipe (QFTLR), a compound of traditional Chinese herbal medicine for dispelling wind and removing obstruction in the meridians, on cell proliferation and expressions of transforming growth factor-beta1 (TGF-beta1) and interleukin-6 (IL-6) mRNAs induced by lippolysaccharide in glomerular mesangial cells from rats. METHODS: The method of serum pharmacology was used. A total of 32 rats were divided into normal control group, untreated group, QFTLR group and positive control group which also was named Monopril (fosinopril sodium) group. Mesangial proliferative glomerulonephritis was induced by injection of antithymocyte serum in the rats except for the normal control group. Sera of the rats were obtained after corresponding interventions. Lipopolysaccharide-induced proliferation of rat mesangial cells (MCs) cultured in the respective serum-containing media was detected by the method of methyl thiazolyl tetrazolium (MTT) assay, and the expressions of TGF-beta1 and IL-6 mRNAs were analyzed by the method of reverse transcription polymerase chain reaction. RESULTS: Compared with the untreated group, QFTLR showed remarkable inhibitory function on the proliferation of the mesangial cells (P<0.05). The expressions of TGF-beta1 mRNA in mesangial cells were increased in the untreated group, QFTLR group and Monopril group when compared with the normal control group (P<0.01), but the TGF-beta1 mRNA expressions in QFTLR group and in Monopril group were lower than that in the untreated group. The IL-6 mRNA expression could be increased by the LPS stimulation, and it was significantly higher in the other three groups than that in the normal control group, including the untreated group, the Monopril group and the QFTLR group (P<0.01). Compared with the untreated group, the expression of IL-6 mRNA was decreased by QFTLR and Monopril (P<0.01). QFTLR was better than Monopril in inhibiting the proliferation of the mesangial cells and decreasing the expressions of TGF-beta1 and IL-6 mRNAs (P<0.05). CONCLUSION: QFTLR has great inhibitory effect on mesangial cell proliferation and expressions of TGF-beta1 and IL-6 mRNAs, which may be one of its mechanisms in postponing glomerular sclerosis.

Clinical study of Trilogy Detoxicating Therapy combined with routine Western medicine on patients with chronic renal failure.

OBJECTIVE: To investigate the mechanism of Trilogy Detoxicating Therapy in treating patients with chronic renal failure (CRF). METHODS: A total of 142 patients were assigned to the Trilogy Detoxicating Therapy group (the treatment group, 82 patients) and the Western medicine treatment group (the control group, 60 patients). All of the patients were treated with NovoNorm 1 mg and metformin hydrochloride tablets 0.15 g thrice per day for lowering the blood glucose, as well as Perindopril 4 mg twice daily for lowering blood pressure, recombinant human erythropoietin 2 000 U and a hypodermic injection thrice a week for rectifying anemia, 30 days as one course of treatment, and all patients were treated for two courses. Patients in the treatment group were treated with the Trilogy Detoxicating Therapy [dispersing the five-zang organs, expelling toxins through colonic dialysis and skin dialysis fumigation] in addition to the aforementioned drugs. Parameters observed and recorded in the study included renal function [serum creatinine (SCr), blood urea nitrogen (BUN)], blood lipids [triglyceride (TG), total cholesterol (TC), low-density lipoprotein C (LDL-C), high-density lipoprotein C (HDL-C)], plasma total protein (TP), hemoglobin (Hb), serum interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha) before and after the treatment. RESULTS: After two courses of treatment, the levels of SCr, BUN, TG, TC, LDL-C, serum IL-6 and TNF-alpha decreased significantly, meanwhile HDL-C increased in the treatment group (P<0.05 or P<0.01). In contrast, no obvious changes of the above mentioned items occurred in the control group. In both groups, the levels of TP and Hb were significantly elevated (P<0.05 or P<0.01), but the changes were more obvious in the treatment group (P<0.01). CONCLUSION: Trilogy Detoxicating Therapy played a therapeutic role on patients with CRF possibly through lowering the levels of blood lipids, serum IL-6 and TNF-alpha.

[Mechanism of the effect of Tongluo Recipe against glomerular sclerosis in rats].

OBJECTIVE: To investigate the effects of Tongluo Recipe on the expression of collagen IV (Col IV), fibronectin (FN), laminin (LN), transforming growth factor-beta1 (TGF-beta1) in rat renal tissues and explore the mechanism underlying these effects in rats with glomerular sclerosis. METHODS: The pathological changes in the renal tissues of rats with glomerular sclerosis were observed microscopically, and the expressions of Col IV, FN, LN, and TGF-beta1 were detected using immunohistochemical staining and image analysis system. RESULTS: Tongluo Recipe significantly decreased the expressions of Col IV, FN, LN and TGF-beta1 in the renal tissue of rats with glomerular sclerosis (P<0.05 or P<0.01) and obviously alleviated the renal pathologies (P<0.01). CONCLUSION: The therapeutic effects of Tongluo Recipe are probably mediated by lowered expressions of Col IV, FN, LN and TGF-beta1.