[Effects of losartan on MT3-MMP and TIMP2 mRNA expressions in diabetic rat kidney].

OBJECTIVE: To investigate the changes of the mRNA expressions of membrane type 3-matrix metalloproteinase (MT3-MMP) and tissue inhibitor of matrix metalloproteinase 2 (TIMP2) in diabetic rat kidneys and the effect of losartan on such changes. METHODS: Three groups of male Wistar rats were used in this study, namely group A as the control group (n=11), group B consisting of diabetic rats without any therapy (n=11), and group C treated with losartan (n=9). Six months after treatment, the kidneys were taken from the rats to measure the mRNA expressions of MT3-MMP, TIMP2 and transforming growth factor beta1eTGFbeta1 with reverse transcriptional PCR (RT-PCR), and observe the glomerular basement membrane thickening and mesangial matrix MMedensity (MM area/mesangial area) with electron microscope. Twenty-four-hour urine was also collected to measure urinary albumin excretion (UAE). RESULTS: The expression of renal MT3-MMP mRNA in group B (1.37+/-0.96) was significantly stronger than those in group A (0.75+/-0.34, P<0.05) and in group C (0.75+/-0.30, P<0.05). The mRNA expression of renal TIMP2 in group B (0.73+/-0.37) was significantly increased as compared with group A (0.32+/-0.19, P<0.05) and group C (0.34+/-0.17, P<0.05), with a higher mRNA expression of renal TGFbeta1 in group B (0.53+/-0.20 vs 0.26+/-0.13 in group A and 0.29+/-0.15 in group C, P<0.05). UAE in group B (2.18+/-1.98 mg) was significantly higher than those in groups A and C (0.41+/-0.47 mg/d, P<0.05; 0.65+/-0.89 mg/d, P<0.05, respectively). The glomerular basement membrane thickness (532+/-108 nm) and the MM density (56.4+/-6.8) were significantly greater in group Bthan in the other two groups (P<0.05). CONCLUSIONS: MT3-MMP and the TIMP2 mRNA expressions are significantly increased in diabetic rat kidneys. Losartan can prevent the development of diabetic nephropathy and inhibit MT3-MMP and the TIMP2 mRNA expressions in diabetic rat kidneys.

Manumycin induces apoptosis in prostate cancer cells.

BACKGROUND: Manumycin exhibits an antitumor effect in a variety of cancer cell lines, including prostate cancer cell lines (DU145 and PC-3). Our previous studies demonstrated that manumycin induced the apoptosis of anaplastic thyroid cancer cells and leukemia cells via the intrinsic apoptosis pathway. In the current study, we further evaluated the effect of manumycin in two prostate cancer cell lines (LNCaP and 22Rv1), and here we elucidate some of the underlying mechanisms. MATERIALS AND METHODS: The cell viability of prostate cancer cells was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay after treatment with manumycin for 48 hours. Apoptosis was detected by flow cytometry using annexin V and propidium iodide. The expressions of B-cell lymphoma (Bcl)-2 family members and the activations of caspase-9 and caspase-3 were detected by Western blotting. RESULTS: Manumycin treatment resulted in significant decreases in the viabilities of the two prostate cancer cell lines in a dose-dependent manner through apoptosis, and this apoptosis involved caspase-9 activation. A specific inhibitor of caspase-9 protected cells from caspase-3 activation, apoptosis, and cytotoxicity induced by manumycin. We also found that manumycin downregulated Bcl-2 expression and upregulated Bax expression. CONCLUSION: Our data suggest that manumycin induces apoptosis in prostate cancer cells through regulation of the Bcl-2 family involving caspase-9 activation. These results suggest that manumycin may be beneficial for the treatment of prostate cancer.

Requirement of reactive oxygen species generation in apoptosis of leukemia cells induced by 2-methoxyestradiol.

AIM: To investigate the effects of 2-methoxyestradiol (2-ME) on 2 myeloid leukemia cell lines HL-60 and U937, and to explore its mechanisms. METHODS: Human myeloid leukemia cells HL-60 and U937 were used. Measurement of mitochondrial membrane potential (Dym) was performed using 5,5',6,6'-Tetrachloro-1,1',3,3'- tetraethylbenzimidazolylcarbocyanine iodide ( JC-1). Apoptosis and cellular nitric oxide (NO) were detected by flow cytometry using Annexin V and NO sensor dye. Superoxide anion was measured with a fluorescent plate reader by dihydroethidium (DHE). Cytotoxicity was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium assay. RESULTS: 2-ME resulted in viability decrease in a dose-dependent manner. 2-ME treatment also generated reactive oxygen species (ROS), including NO and superoxide anions, which resulted in mitochondria damage. 2-ME-induced apoptosis was correlated with an increase in ROS. The quenching of ROS with N-acetyl-L-cysteine protected leukemia cells from 2-ME cytotoxicity and prevented apoptosis induction by 2-ME. Furthermore, the addition of manumycin, a farnesyltransferase inhibitor, significantly enhanced apoptosis induced by 2-ME. CONCLUSION: Cellular ROS generation plays an important role in the cytotoxic effect of 2-ME. It is possible to use ROS generation agents, such as manumycin, to enhance the antileukemic effect. The combination strategy needs further in vivo justification and may have potential clinical application.

[Clinical features and prognosis of IgA nephropathy with glomerular crescents].

OBJECTIVE: To understand the clinical features and prognosis of IgA nephropathy (IgAN) patients with glomerular crescents. METHODS: The clinical data collected at the time of renal biopsy and the follow-up data of 89 IgAN patients with glomerular crescents were analyzed with 412 IgAN patients without crescents serving as the control group. Follow-up study was conducted in 78 patients with crescents and 198 without it, and the renal survival rate was estimated using Kaplan-Meier survival analysis. RESULTS: The incidence of glomerular crescents was 17.8% in IgAN patients. Clinically, 39 patients with crescents experienced rapidly progressive glomerulonephritis, resulting in a significantly higher rate of this manifestation than that in patients without crescent. Patients with crescents also had more grave pathological changes in the glomerulus and renal tubule interstitium than the control patients. Patients were followed up for an average of 40.3-/+29.6 months in crescent group and 45.1-/+26.9 months in the control group, and the 1-, 3-, 5-year renal survival rate was 95.24%, 80.95%, 61.9% in the former and 100%, 91.67%, 79.17% in the latter, respectively. CONCLUSION: IgAN patients with crescents have severer clinical and pathological manifestations and poorer prognosis than those without crescents.