ABSTRACT
<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>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.</p>
Subject(s)
Animals , Rats , Cell Line , Cyclin D1 , Metabolism , Cyclin-Dependent Kinase 2 , Metabolism , Cyclin-Dependent Kinase Inhibitor p21 , Metabolism , Cyclin-Dependent Kinase Inhibitor p27 , Metabolism , Drugs, Chinese Herbal , Pharmacology , Gene Expression Regulation , Mesangial Cells , Cell Biology , Metabolism , Rats, Sprague-DawleyABSTRACT
Objective To investigate the expression of anionic site in adriamycin-induced nephropathy (AN) rats, and to further explore the effects of Qufengtongluo Recipe on charge barrier in glomerulus in AN rats. Methods Adriamycin nephropathy was induced by a single tail intravenous injection of adriamycin. Totally 80 rats were randomly divided into normal control group and nephropathy model group. Three weeks later, the nephropathy model was established, and 50 AN rats were randomly divided into five groups: nephropathy model group (B, n=10), Qufeng group (C, n=10), prednisone and Qufeng group (D, n=10), prednisone group (E, n=10) and benazepri group (F, n=10), and they were treated respectively. With treatment being given respectively, renal tissue samples in each group were collected at week 3 and 7, respectively. The ultrastructure and expression of anionic site were examined by electron microscope observation. Results ① After adriamycin injection, a significant increase of the 24-hour urinary protein was observed at week 3 (P<0.01). In AN rats, serum albumin was decreased (P<0.01) while serum TCH and TG were increased (P<0.01). ② In AN rats the diffuse fusion and effacement of foot processes and decrease of anionic sites in GBM were observed at week 3. ③ At week 7, the average intensity of AS dramatically increased in C and E groups (P<0.01) compared with that in nephropathy model group. Conclusion The abnormal expression of AS is the important mechanism that leads to the occurrence and development of proteinuria in AN rats. It is possible that Qufengtongluo Recipe has effects on nephrotic syndrome through altering the charge barrier in GBM in glomerulus.
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Objective To explore the effects of Yishen Capsule on the soluble endothelium protein C receptor (sEPCR) and P-selectin (CD62P) as well as the serum levels of immunoglobulin (Ig) and complement 3 (C3) in patients with chronic glomerulonephritis (CGN). Methods The changes of sEPCR and CD62P levels in plasma as well as Ig and C3 levels in serum were assessed in 40 normal subjects and 78 patients before and after treatment by double-antibody enzyme-linked immunosorbent assay, flow cytometry and immunoturbidimetry, respectively. Results Compared with those in normal group before treatment, the levels of sEPCR and CD62P in plasma as well as IgA in serum increased obviously (P<0.01), and the levels of IgG and IgM as well as C3 in serum decreased significantly in CGN patients (P<0.01). After three months' treatment with Yishen Capsule, the levels of sEPCR and CD62P in plasma and IgA in serum decreased significantly (P<0.05 or P<0.01), and the levels of IgG and IgM in plasma as well as C3 in serum increased obviously (P<0.05 or P<0.01). Conclusion The mechanism of Yishen Capsule's effect in treating patients with CGN may be correlated with decreasing sEPCR and CD62P levels in plasma as well as regulating the levels of Ig in serum and C3.
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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.
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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.
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Clinical practice has proved that regulating activities ofqi and recovering function ofthe liver in governing conveyance and dispersion, can make the chronic kidney disease(CKD) patients' symptoms improved actually.In this paper, we explore the ‘treatment ofchronic kidney disease through the liver' from the aspects ofanalyzing the relationship ofthe liver wood and kidney water's physiology and pathology, the theoretical origin ofthe treatment ofkidney disease through the liver, the liver and multi-system ofmodern medicine and so on.‘Treatment ofchronic kidney disease through the liver' is to apply regulating activites ofqi throughout the whole treatment course.It can help the liver restore the balance ofvarious systems steady state, the internal environment stability, so as to protecting the kidney function.It belongs to harmonizing therapy, the clinical specific prescription is Xiaochaihu Tang