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.

A clinical randomized controlled trial: moxibustion at Laogong interval with Panax notoginseng promoted the maturation of arteriovenous fistulae.

BACKGROUND: We aim to study the clinical effect of moxibustion at Laogong interval with Panax notoginseng on the short-term maturation and long-term patency of arteriovenous fistula. METHODS: Seventy-four pre-dialysis uremic patients who received distal forearm radial-cephalic fistula creations were enrolled in this study and randomly assigned to the control group and experimental group. After arteriovenous fistula creations, the control group underwent handgrip exercise, and the experimental group received moxibustion at Laogong acupoint interval with Panax notoginseng. Both groups received a 12-week treatment and were followed up for 24 weeks in all at the following time points: before creations and 2, 4, 8, 12, 24 weeks after creations. The diameter of anastomosis, the diameter and outflow of draining-veins 5 cm above anastomosis, the diameter and outflow of brachial arteries evaluated the maturation and patency of arteriovenous fistula. Enzyme linked immunosorbent assay determined serum levels of endothelin and nitric oxide. RESULTS: The maturity rate in the experimental group was significantly higher than that in the control group at 4 weeks after arteriovenous fistula creations (P = 0.048). The diameter of anastomosis, the diameter of draining veins, and the blood flow of draining veins increased in both groups during the whole 24 weeks. The diameter and blood flow of brachial arteries ascended in both groups during the previous 12 weeks. Compared with the control group, moxibustion at Laogong interval with Panax notoginseng significantly improved the value of the diameter of draining-veins (P = 0.016), the blood flow of draining-veins (P = 0.015), the diameter of brachial arteries (P < 0.001), and the blood flow of brachial arteries (P = 0. 012) at 2 weeks, and enhanced the blood flow of draining-veins (P = 0.029) and brachial arteries (P < 0.001) at 12 weeks. Serum levels of endothelin were significantly lower (P = 0.047), and serum levels of nitric oxide were markedly higher (P < 0.001) in the experimental group than that in the control group at 2 weeks after creations. CONCLUSIONS: Moxibustion at Laogong interval with Panax notoginseng was non-invasive and promoted the maturation of arteriovenous fistula at 4 weeks after creations. However, its long-term beneficial effect on patency at 24 weeks after creations was not significant. Trial registration Chinese Clinical Trial Registry, No. ChiCTR1900024042. Registered, http://www.chictr.org.cn/index.aspx.

Reactive oxygen species induced by uric acid promote NRK­52E cell apoptosis through the NEK7­NLRP3 signaling pathway.

Increasing uric acid (UA) could induce renal tubular epithelial cell (NRK­52E) injury. However, the specific mechanism by which UA induces renal tubular epithelial cell injury remains unknown. It was hypothesized that UA induces renal tubular epithelial cell injury through reactive oxygen species (ROS) and the Never in mitosis gene A (NIMA)­related kinase 7 (NEK7)/NLR family pyrin domain containing 3 (NLRP3) signaling pathway. TUNEL assay and flow cytometry were applied to measure apoptosis, and the results of the present study showed that UA treatment induced apoptosis of NRK­52E cells in a concentration­dependent manner. Western blotting was performed to determine the expression levels of cleaved caspase­3, Bax and Bcl­xl, it was found that levels were significantly increased after UA treatment in NRK­52E cells. ROS and apoptosis were predominantly induced in NRK­52E cells and there was an association between ROS and apoptosis. Enhanced expression of NEK7, NLRP3, apoptosis­associated speck­like and caspase­1 were observed in NRK­52E cells treated with UA. The ROS inhibitor, N­acetyl­l­cysteine, exerted a protective effect on the UA­induced apoptosis of tubular epithelial cells by reducing excess ROS production, which significantly inhibited NEK7 and NLRP3 inflammasome activation. These results indicated that UA activates ROS and induces apoptosis of NRK­52E cells. The mechanism might be related to the regulation of the NEK7/NLRP3 signaling pathway.

[Effect of fuzheng huayu recipe and huangqi tang on DMN-induced experimental liver cirrhosis in rats].

OBJECTIVE: To investigate the effects of Fuzheng Huayu recipe and Huangqi tang on DMN-induced experimental liver cirrhosis in rats and explore the therapeutic characteristics of Buxu herbals on liver cirrhosis. METHOD: Liver cirrhosis in rats was induced by intraperitoneally injection of DMN for 4 weeks. Cirrhotic rats were randomly divided into 4 groups: model group, and Fuzheng Huayu recipe group, Huangqi tang group, Fuzheng Huayu recipe combined with Huangqi Tang group. The rats in treatment groups were orally administered with Fuzheng Huayu recipe, Huangqi tang, Fuzheng Huayu recipe combined with Huangqi tang (1:1), respectively. Normal and model control rats were given the equivalent normal saline. The body weight, liver weight and spleen weight were observed when rats were sacrificed. Liver histology was examined by HE staining and Sirius red staining. The liver function parameters including ALT, T. Bil and Alb were determined. The SOD activity and MDA content in liver tissues were also measured. Hepatic hydroxyproline (Hyp) content was determined by Jamall's method. The expression of alpha-SMA was determined by both immunohistochemistry staining and western blot method. RESULT: Compared with normal rats, the serum ALT and T. Bil levels in model rats increased obviously, by contrast, the serum Alb level decreased. There was a significant decline of SOD activity in model rat liver tissue, while the content of MDA and Hyp increased remarkably. A severe deterioration of liver architecture, infiltration of inflammatory cells and deposition of collagen were observed in model rat liver tissue. The expression of alpha-SMA also increased significantly. Compared with model rats, the liver function, lipid peroxidation parameters, Hyp content and liver histology were all improved in the 3 treatment groups. The combined group is better than any single-use group in decreasing collagen deposition and expression of alpha-SMA. CONCLUSION: Fuzheng Huayu recipe, Huangqi tang, Fuzheng Huayu recipe combined with Huangqi tang can attenuate liver fibrosis in DMN induced rats. Fuzheng Huayu recipe combined with Huangqi tang is better than that using alone in decreasing collagen deposition. The mechanism is partially due to the better effect of Fuzheng Huayu recipe combined with Huangqi tang on inhibiting activated HSC.