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@#AIM: To evaluate the effect of leech extract on human retinoblastoma WERI-RB-1 cells.<p>METHODS: WERI-RB-1 cells with different concentrations of leech extracts(0.02, 0.04, 0.08 and 0.16U/mL)were cultured in vitro for 0, 24, 48 and 72h. The optimal drug intervention concentration and time were selected by CCK-8 method for subsequent experiments. WERI-RB-1 cells cultured <i>in vitro</i> were divided into control group(normal culture medium)and experimental group(culture medium containing leech extract). Flow cytometry was used to detect the effects of drugs on cell cycle and cell apoptosis, and Transwell invasion assay was used to detect the effects of drugs on cell invasion abilit. <p>RESUITS: According to the detection results of CCK-8 method, the optimal intervention conditions were 0.04 and 0.08U/mL leech extract for 48h. The cells in the leech extract intervention were mainly blocked at G2/M stage, and the positive cell rates in the 0.04 and 0.08U/mL groups were(12.59±5.36)% and(14.79±4.12)%, respectively, which were significantly higher than those in the control group \〖(3.00±2.32)%, <i>P</i><0.01\〗. The apoptosis rate of the cells in the 0.04 and 0.08U/mL groups was(37.91±3.44)% and(33.05±2.25)%, respectively, which were significantly higher than that in the control group \〖(4.64±2.56)%, <i>P</i><0.01\〗. The results of Transwell invasion assay showed that the number of cells under the Transwell chamber in the experimental group was significantly than that in the control group, indicating that the leech extract could inhibit cell invasion.<p>CONCLUSION: The leech extract can inhibit the proliferation and invasion of human retinoblastoma cells and induce cell apoptosis <i>in vitro</i>.
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Objective To study the inhibition effect of leech extract on HepG2 cells. Methods Human hepatocellular cancer cell line HepG2 were treated with different concentrations of leech extracts which were extracted by method of freeze-thawing with liquid nitrogen and contrasted with that by method of water extracting and ethanol precipitating. The inhibition effects and cell morphous were examined by MTT assay and Acridine orange (AO) fluorescent staining method respectively. Result The 6～15 mg/mL drug concentrations of leech extract by method of freeze-thawing with liquid nitrogen had an obvious inhibition on proliferation of HepG2 cells in a dose-dependent manner, and the effect was better than that by method of water extracting and ethanol precipitating (P
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Background: The vosoactive effects of proteins isolated from leech saliva and leech homogenate have been the focus of many recent studies because of its therapeutic potential. Possible nonlethal and lethal toxic effects have to be determined before any therapeutic effects could be tested. The general objective is to take the first step in drug design by performing an acute toxicity studyMethods: In this randomized, double-blind trial, Swiss mice were administered subcutaneously with varying doses of leech Hirudinaria manillensis (Lesson) homogenate and observed for possible acute toxicity. Toxidromes appearing within the two-week period after the administration of the substance were noted. Necropsy was performed on all the mice subjects. The LD(50) was computed using the log dose-response probit analysis. The doses (in g/kg body weight) were 3.54, 5.0 and 7.06Results: The LD(50) at Day 2 of Hirudinaria manillensis extract was 4.6124 g/kg body weight. The LD(50) at Day 14 was not obtained, because the test animals incurred mortalities beyond Day 2, which could be explained by a delayed toxicity of the test substance. Mice injected with 7.06 g/kg showed the most number of observable toxidromes, involving several organ systems, which generally had an early onset and persisted until the time of death of the mice. Mice treated with 3.54 g/kg showed toxidromes with early onset time, but they were not as consistent as the highest dose and the effects were generally not long-lasting. Motor activity was the most affected among the toxidromes. Gross pathologic findings revealed that the intestine, liver, heart, stomach, lungs and kidney are the commonly affected organs, which had marked changes in all dose groupsConclusions: The crude H. manillensis leech extract caused toxicity in mice, with toxidromes involving mainly the nervous system, specifically the somatosensory and neuromuscular systems. The LD(50) could not be determined in this study. However, using the data at Day 2, LD(50) was 4.6124 g/kg body weight. The LOAEL could be less than or equal to 3.54 g/kg body weight. No observable adverse effect level could not be determined. (Author)