[Effects of ferulic acid on inflammation and autophagy levels in glomerular mesangial cells induced by high glucose].

OBJECTIVE: To investigate the protective effects and possible mechanisms of ferulic acid on diabetic nephropathy by observing the effects of ferulic acid on the level of inflammation and autophagy in glomerular mesangial cells induced by high glucose. METHODS: SV40 MES 13 cells were cultured and randomly divided into the following groups: normal group (Control, 5.6 mmol/L glucose), mannitol group (Man, 30 mmol/L mannitol), high glucose group (HG, 30 mmol/L glucose), ferulic acid group (FA, 30 mmol/L glucose + 12.5, 25, 50, 100, 200 µmol/L ferulic acid), and the proliferation of SV40 MES 13 cells in each group was observed by MTT method. The levels of tumour necrosis factor-α (TNF-α), monocyte chemotactic protein-1 (MCP-1) and interleukin 1ß(IL-1ß)in cell supernatant were determined by enzyme-linked immunosorbent assay (ELISA). The expressions of NLRP3, IL-1ß, LC3-II/I and p62 proteins in SV40 MES 13 cells were detected by Western blot. RESULTS: ①The proliferative activity of SV40 MES 13 cells was significantly higher in the HG group compared to the control group (P＜0.01), while the proliferative activity of SV40 MES 13 cells was decreased to different degrees in the FA group compared to the HG group (P＜0.05～0.01). ②Compared to the control group, the levels of TNF-α, MCP-1 and IL-1ß were increased significantly in the cell supernatant of HG group (P＜0.01). Compared with the HG group, the levels of TNF-α, MCP-1 and IL-1ß were decreased significantly in the FA group (P＜0.01). ③Compared with the control group, LC3-II/Ⅰ protein expression was decreased in the HG group, while the levels of p62, NLRP3 and IL-1ß protein were increased significantly (P＜0.01). Compared with the HG group, the expression of LC3-II/Ⅰ protein was elevated significantly (P＜0.05) in the FA group, while the levels of p62, NLRP3 and IL-1ß protein in the FA group were decreased significantly (P＜ 0.01). CONCLUSION: FA can inhibit the abnormal proliferation of SV40 MES 13 cells induced by high glucose. FA can protect glomerular mesangial cells by inhibiting inflammation and increasing the level of autophagy.

[Effects of ketoconazole on physiological function of liver and testis in mice].

Objective: To investigate the effects of different doses of ketoconazole (KCZ) on the physiological functions of the liver and testis in Kunming mice. Methods: Forty male Kunming mice were randomly divided into four groups (n=10): normal group, KCZ low-dose group (30 mg/kg), medium-dose group (50 mg/kg), and high-dose group (70 mg/kg). The mice in the drug groups were injected subcutaneously (0.1 ml/10 g) with the corresponding dose of KCZ once a day, and the concentrations of KCZ in the KCZ low, middle, and high dose groups were 3 mg/ml, 5 mg/ml and 7 mg/ml respectively, and the normal group was injected with the same amount of normal saline for 3 weeks. The activities of aspartate transaminase (AST) and alanine aminotransferase (ALT) in serum, and γ-glutamyl transpeptidase (γ-GT), lactate dehydrogenase (LDH) and acid phosphatase (ACP) in testicular tissue were measured. HE staining was used to observe the pathological changes of the liver and testis. Results: Compared with the normal group, the activities of AST and ALT were increased significantly (P＜0.01), and the activities of γ-GT, ACP and LDH were decreased markedly in KCZ groups (P＜0.01). KCZ could affect the above indexes in a dose-dependent manner. HE staining showed that the hepatocytes were denatured, arranged loosely, and the cytoplasm was light in color. The lumen of the seminiferous tubules of the testis were enlarged, and the number of spermatogenic cells and sperm at all levels were decreased. Conclusion: KCZ could cause physiological function damage and pathological histological changes of the liver and testis, increase the levels of liver transaminase, reduce the activities of testicular specific enzymes of mice. Besides, the degree of damage was increased with the increase of dose.

Icariin ameliorates streptozocin-induced diabetic nephropathy through suppressing the TLR4/NF-κB signal pathway.

Diabetic nephropathy (DN) is one of the complex and severe complications of diabetes mellitus (DM). Icariin (ICA) is a flavonoid extracted from the leaves and stems of Herba epimedii with a wide range of pharmacological effects, such as anti-osteoporosis, anti-fibrosis, anti-aging, anti-inflammation and antioxidation. The purpose of our study was to explore the renal protective effect of ICA on DN in mice and its possible mechanisms. ICR mice were exposed to STZ-induced DN. The kidney organ coefficient of mice was computed. 24 h UP in urine was measured. Serum FBG, Cr and BUN were detected. The content of MDA and the activities of SOD, CAT and GSH-Px in renal tissues were tested. HE staining, PAS staining, PASM staining and transmission electron microscopy were used to observe renal pathological changes. Furthermore, TLR4, p-NF-κB p65, TNF-α and IL-6 of renal tissues were assayed by immunohistochemistry and western blotting. Our results indicated that ICA observably optimized the renal organ coefficient, reduced the level of 24 h UP in urine, decreased the content of Cr, BUN in serum and MDA in renal tissues, promoted the activities of SOD, CAT and GSH-Px in renal tissues, and ameliorated pathological lesions of kidneys noticeably. Besides, ICA inhibited the expressions of TLR4, p-NF-κB p65, TNF-α and IL-6 remarkably in renal tissues. ICA, which might lighten the renal inflammatory response by suppressing the TLR4/NF-κB signal pathway, played a protective role in kidneys of STZ-induced DN mice.