OBJECTIVE: To investigate the effects of Shenshuning Recipe (SSNR) on gene expression of hepatocyte growth factor (HGF) in renal tissues in rats with glomerulosclerosis. METHODS: Glomerulosclerosis was induced in 42 rats by unilateral nephrectomy and intravenous injection of doxorubicin. Then these 42 rats were randomly divided into three groups: untreated group, SSNR-treated group and benazepril-treated group. Another eight rats were included into sham-operation group. The rats in the SSNR-treated group and the benazepril-treated group were fed SSNR or benazepril respectively for 8 weeks. The levels of 24 h urine protein (Upr), serum creatinine (Cr) and blood urea nitrogen (BUN) of rats in each group were examined. The renal morphological changes were observed under microscope, and the diameter of glomerular capillary, mesangial matrix and glomerulosclerosis index were analyzed by image analysis software. Reverse transcription-polymerase chain reaction (RT-PCR) method was used to detect the gene expression of HGF in the renal tissues. RESULTS: The levels of 24 h Upr, serum Cr and BUN in the untreated group were remarkably increased than those in the sham-operation group (P<0.01). The pathological morphological changes in the untreated group showed that the glomerulosclerosis was diffused around the renal tissue and the capillaries were shrunk. The expression level of mesangial matrix was up-regulated and the glomerulosclerosis index was 3.32+/-0.35. The expression level of HGF mRNA in the untreated group was obviously lower than that in the sham-operation group (P<0.05). The levels of 24 h Upr, serum Cr and BUN in the SSNR-treated group and the benazepril-treated group were remarkably decreased as compared with those in the untreated group, while the expression levels of HGF mRNA were both obviously higher than that in the untreated group (P<0.01). The pathological morphological changes in the SSNR-treated group and the benazepril-treated group were both alleviated. There was no significant difference in therapeutic effect between the SSNR-treated group and the benazepril-treated group. CONCLUSION: Shenshuning Recipe can up-regulate the expression of HGF mRNA, decrease the mesangial matrix, and improve the renal function, so that it may retard the development of glomerulosclerosis.

Objecfive To investigate the change of V-ATPase B subunits on epithelial to mesenchymal transition (EMT)in rat renal tubular epithelial cells (NRK52E) stimulated by transforming growth factor β1 (TGF-β1). Methods NRK52E cells were stimulated by TGF-β1 (10 μg/L)for O h(control),12 h,24 h,48 h,72 h after sefrum-free culture for 24 h.The mRNA and protein expression of E-cadherin,α-SMA,B2 and B1 subunits of V-ATPase were detected by real-time PCR,Western blotting and immunofluorescence. Results After stimulated by TGF-β1 (10 μg/L)for 48 h,the expression of α-SMA was markedly increased(P<0.05),but the expression of E-cadherin was dramatically decreased(P<0.05).Meanwhile,the expressions of V-ATPase subunit B2 was significantly increased (P<0.05).However,the B1 subunit distributed rarely in NRK 52E cells,and did not increase after TGF-β1 stimulation.Double-label immunofluoerscence staining also showed that the V-ATPase B2 subunit was increased in the cytosol.tending to accumulate to the cell membrane after TGF-β1 stimulation. Conclusions The main isoform of V-ATPase distributed in NRK52E cells is B2 subunit.B2 subunit is increased alone with TGF-β1-induced EMT.It may suggest that V-ATPase B2 subunit may play a potential role in TGF-β1-induced tubular EMT and renal fibrosis.

Objective: In this study, black tea and Citrus maxima (BT-CM), yellow tea and C. maxima (YT-CM), green tea and C. maxima (GT-CM) as subjects, the active ingredient content and antioxidant activity of three tea and C. maxima (T-CM) were analyzed. The effects of three T-CMs on apoptosis of liver cells in vitro and its mechanism were further explored. Methods: National standard method and HPLC were used for active ingredient analysis. MTT, cell flow cytometry and Western blot were used to analyze the effects of three T-CMs on cell proliferation, apoptosis, and its underlying molecular mechanism. Results: The content of tea polyphenols, free amino acids, ratio of polyphenols and amino acids, ester catechins, non-ester catechins and caffeine in YT-CM and GT-CM was significantly higher than that of BT-CM. The in vitro antioxidant capacity of YT-CM and GT-CM was also significantly stronger than that of BT-CM. Three T-CMs had the effects of inhibiting proliferation, arresting cell cycle and inducing apoptosis in HepG2 and Bel7402 cells, especially YT-CM and GT-CM. Western blot analysis showed three T-CMs activated PI3K/AKT/mTOR signaling pathway and regulated the expression levels of apoptosis-related proteins Bax, Bcl-2 and Caspase-3/9. YT-CM and GT-CM had better ability to change the signal pathway than BT-CM. Conclusion: In short, T-CMs, which combined different degrees of fermentation tea with C. maxima, were rich in nutrients and biologically active substances. T-CMs, especially YT-CM and GT-CM, are healthy drinks that help to prevent and treat liver cancer.