Guben Tongluo Formula Protects LPS-induced Damage in Lamina Propria B Lymphocytes Through TLR4/MyD88/NF-κB Pathway.

OBJECTIVE: The main pathological feature of immunoglobulin A nephropathy (IgAN), an autoimmune kidney disease, is the deposition of IgA immune complexes, accompanied by mesangial cell proliferation and elevated urine protein. The Guben Tongluo formula (GTF) is a traditional Chinese medicine prescription, which has predominant protective effects on IgAN. However, the therapeutic mechanism of the GTF in IgAN remains elusive. The present study aimed to determine the effects of GTF in treating IgAN via regulating the TLR4/MyD88/NF-κB pathway. METHODS: In the present study, lamina propria B lymphocytes were treated with different concentrations of lipopolysaccharide (LPS) (0, 1, 5, 10 and 20 ng/mL). Flow cytometry was used to define positive CD86+CD19+ cells. CCK-8 assay was used to examine cell proliferation. RNAi was used to induce TLR4 silencing. qRT-PCR and Western blotting were used to determine gene expression. RESULTS: It was found that the LPS dose-dependently increased the content of IgA and galactose-deficient IgA1 (Gd-IgA), the levels of TLR4, Cosmc, MyD88 and phosphorylated (p)-NF-κB, and the ratio of CD86+CD19+ and IgA-producing B cells. However, the TLR4 knockdown reversed the role of LPS. This suggests that TLR4 mediates the effects of LPS on lamina propria B lymphocytes. Furthermore, the GTF could dose-dependently counteract the effects of LPS and TLR4 overexpression on lamina propria B lymphocytes through the TLR4/MyD88/NF-κB pathway. CONCLUSION: Collectively, these results demonstrate that the GTF can regulate the TLR4/MyD88/NF-κB pathway to treat IgAN model lamina propria B lymphocytes stimulated by LPS.

[Effects of Gubentongluo Formula on Oxidative Stress Reflected by Expressions of PPARα and L-FABP in Mice with IgA Nephropathy].

OBJECTIVES: To determine the underlying mechanism of Gubentongluo Formula in the treatment of IgA nephropathy (IgAN). METHODS: C57BL/6 mice were randomly divided into four groups: normal group (n =10), IgAN group (n =10), control group (n =10) and treatment group (n =10). Mice in the normal and IgAN groups were intragastricly administered with normal saline for 12 weeks; while those in the control and treatment groups were given fenofibrate [30 mg/(kg!$d) and Gubentongluo Formula [1.67 mL/(g!$d)], respectively. Urinary albumin was detected at week 0 and 12. At week 12, protein expressions of peroxisome proliferstor activated receptor α (PPARα), liver fatty acid-binding proteins (L-FABP), 4-hydroxy-2-nonenal (4-HNE), and hemeoxygenase-1(HO-1) in renal tissues were determined by Western blot; mRNA expressions of PPARα and L-FABP in renal tissues were determined by florescent quantitative reverse transcription-polymerase chain reaction (qRT-PCR). RESULTS: At week 12, higher levels of urinary albumin, pathological injuries in glomerular mesangial area, and lower expressions of protein and mRNA of PPARα and L-FABP were found in mice in the IgAN group compared with those in the normal group (P <0.01). The levels of those indicators decreased in those treated with fenofibrate and Gubentongluo Formule, but still higher than the normal controls (P <0.01). The mice treated with Gubentongluo Formula had more significant improvement than those treated with fenofibrate (P <0.05). CONCLUSION: [CM(155.3mm]Gubentongluo formula can improve proteinuria and pathological injuries in glomerular mesangial area of IgAN mice, due to reduction of oxidative stress in renal tissues through regulating the expressions of PPARα and L-FABP.

[Effect of "Gubentongluo Formula" on the IgA Class Switch Recombination of B Lymohocytes in Peyer's Patches in Mice with IgA Nephropathy].

OBJECTIVE: To explore the underlying mechanism of "Gubentongluo Formula" in treatment of IgA nephropathy (IgAN). METHODS: After the IgAN model was successfully induced at week 12, the Kunming mice were randomly divided into three groups: normal control group (n = 15), IgAN group (n = 15) and Traditional Chinese Medicine (TCM) group. The mice in normal control and IgAN group were intragastriclly administrated with normal saline for 8 weeks; meanwhile, the mice in TCM group were intragastriclly administrated with "Gubentongluo Formula" 1.35 mL/ (g · d). The levels of 24 h urine protein were determined at Week 0, 12 and 20. At week 20, the changes of renal pathology were detected; the mRNA expressions of transforming growth factor-ß (TGF-ß) and small mothers against decapentaplegic (Smad) 3 in Peyer's patches (PPs) were detected by fuorescent quantitative reverse transcription-polymerase chain reaction; the protein expressions of TGF-ß and Smad 3 in PPs were detected by immunohistochemistry technique; the levels of (IgA + B)/B lymphocytes in PPs were determined by flow cytometry. RESULTS: Compared with those results of normal control group, the levels of 24 h urine protein, IgA deposition in glomerular mesangial area, and expressions of protein and mRNA of TGF-ß and Smad3 in IgAN group were significantly increased (P < 0.01). Besides, the levels of (IgA+B)/B lymphocytes were significantly elevated in IgAN group (P < 0.01). All these indicators were improved in TCM group. Compared with IgAN group, the differences were statistically significant (P < 0.01). Compared with those results of control group, the levels of (IgA + B)/B lymphocytes showed no significant difference in TCM group (P > 0.05), but other indicators showed significant differences (P < 0.01). CONCLUSION: "Gubentongluo Formula" could effectively improve proteinuria and suppress IgA deposition in glomerular mesangial area in IgAN mice, due to affect IgA class switch recombination of B lymphocytes in PPs through regulating TGF-ß/Smad3 pathway.

Renal protection of losartan 50 mg in normotensive Chinese patients with nondiabetic chronic kidney disease.

AIMS: Nondiabetic chronic kidney disease (CKD) is the leading major cause of end-stage renal disease in developing countries including China. Among the 5 stages of CKD, it is critical to retard the progression of stage 3 because renal disorder could accelerate aggravation behind that stage. Data suggest that high dosages of angiotensin receptor blockers (ARBs) could retard the progression of renal disease in hypertensive and/or diabetic patients. Nevertheless, in daily practice of nephrology, quite a number of nondiabetic patients with CKD who are normotensive do not tolerate even moderate dosages of ARBs because of adverse effects such as systemic hypotension, epically for Chinese patients. The aim of this study was to investigate the renoprotective effects of relatively low dosages of ARBs in normotensive Chinese patients with nondiabetic stage 3 CKD. METHODS: A prospective, randomized, parallel-group, open-label study was performed over a period of 12 months. A total of 238 enrolled patients were randomly allocated to treatment with losartan 50 mg (n = 119) or placebo (n = 119). All patients were followed up at 2-month intervals. At each visit, blood pressure was measured, and urinalysis and serum biochemistry tests were performed. RESULTS: Finally, 112 patients given losartan and 114 patients given placebo completed the study. In the losartan group, there was a significant and biphasic time-dependent decline in proteinuria during therapy (1.72 ± 0.47 to 0.99 ± 0.48 g/d; P < 0.001). Conversely, placebo did not simultaneously change the amount of proteinuria (1.73 ± 0.49 to 1.64 ± 0.50 g/d; P = 0.337). Estimated glomerular filtration rate remained stable during the entire study period in the patients given losartan (44.8 ± 8.1 to 44.1 ± 7.7 mL/min per 1.73 m; P = 0.120) but were significantly reduced in the placebo group (44.5 ± 8.5 to 39.1 ± 7.4 mL/min per 1.73 m, P < 0.001). The changes in blood pressure were kept constant in the 2 groups. All adverse events were minimal and nonfatal. CONCLUSIONS: For normotensive patients with nondiabetic stage 3 CKD, therapy with a daily dose of losartan, 50 mg, may perform effective renoprotection without changing blood pressure and be generally safe and well tolerated.