铅和（或）镉对体外培养肾细胞的毒性损伤

在培养液中加入不同浓度的Cd（10、20、40μmol／L）、Pb（10、20、40μmol／L）和Pb-Cd联合（分别为5、10、20μmol／L），来探讨铅镉对肾细胞的作用及可能机制。在显微镜下观察细胞形态、测定细胞凋亡率和细胞周期及上清液脂质过氧化指标。结果表明：20μmol／L以上染毒组细胞皱缩、体积变小，表面有细胞碎片，甚至呈泡沫状或树枝状；铅、镉单独和联合染毒组GSH—Px、SOD活性随染毒剂量的增加，呈逐渐下降趋势（P〈0．05）．而MDA含量和凋亡率则呈升高趋势（P〈0．05），并存在剂量和时间效应。镉和铅也能使细胞周期停滞。Pb、Cd联合可加剧细胞损伤。     

白藜芦醇通过激活去乙酰化酶1/腺苷一磷酸激活的蛋白激酶信号通路影响牛脂肪细胞凋亡

本试验旨在研究植物提取物白藜芦醇(RES)对牛皮下脂肪细胞凋亡率以及去乙酰化酶1(SIRT1)/腺苷一磷酸激活的蛋白激酶(AMPK)信号通路关键因子的mRNA和蛋白质表达量的影响。选取18月龄鲁西黄牛的皮下前体脂肪细胞,在细胞分化的第0天,更换为RES浓度分别为0(对照)、100、200和400μmol/L的培养液处理48 h,每组设3个重复。应用Hoechst33342染色检测细胞凋亡的形态学变化,流式细胞分析仪检测细胞凋亡率,荧光定量PCR(q PCR)和免疫印迹试验(Western-blot)分别检测SIRT1/AMPK信号通路上的关键因子SIRT1、A MPKα、叉头转录因子1(Fox O1)、B细胞淋巴瘤/白血病-2(Bcl-2)、半胱氨酸天冬氨酸蛋白酶-3(caspase-3)、促凋亡蛋白Bcl-2相关X蛋白(Bax)的mRNA和蛋白质表达量,并利用油红O染色鉴定脂肪细胞。结果表明,与对照组相比:不同浓度RES处理后的牛皮下脂肪细胞凋亡率均极显著增高(P0.01);不同浓度RES处理后的SIRT1、AMPKα、caspase-3、Bax的mRNA表达量均显著或极显著提高(P0.05或P0.01),Bcl-2的mRNA表达量极显著降低(P0.01);不同浓度RES处理后的SIRT1、AMPKα、caspase-3的蛋白质表达量均显著或极显著提高(P0.05或P0.01);200和400μmol/L RES处理后,Fox O1的mRNA和蛋白质表达量显著或极显著提高(P0.05或P0.01),Bax的蛋白质表达量极显著提高(P0.01),Bcl-2的蛋白质表达量极显著降低(P0.01)。100μmol/L RES处理后时,Fox O1的mRNA和蛋白质表达量以及Bcl-2、Bax的蛋白质表达量均与对照组差异不显著(P0.05)。由此可见,RES通过激活SIRT1/AM PK信号通路,同时激活通路下游的Fox O1,促进了牛皮下脂肪细胞的凋亡,为通过营养调控技术降低牛皮下脂肪沉积提供了一定的理论基础。     

Liraglutide ameliorates liver injury of type 2 diabetic mice via micro-RNA-33-AMPK pathway

AIM: To observe the effects of liraglutide on the level of microRNA-33 (miR-33) and the expression of AMP-activated protein kinase (AMPK) and apoptosis-related proteins in mice with type 2 diabetes mellitus (T2DM), and to explore its possible mechanism. METHODS: High-fat diet and intraperitoneal injection of streptozocin were used to establish the type 2 diabetic model in C57BL/6 mice. The mice were randomly divided into 4 groups (n=15):in control group, the normal mice were subcutaneously injected with equivalent volume of saline; in model group, the T2DM mice were subcutaneously injected with equivalent volume of saline; in low-and high-dose liraglutide treatment groups, the T2DM mice were subcutaneously injected with 100 and 200 μg·kg^-1·d^-1, respectively. After 4 weeks of administration, the levels of FBG, TG, TC, HDL-C, LDL-C, ALT and AST were determined. HE staining was used to observe the pathological changes of the liver tissues. The protein level of cleaved caspase-3 in the liver tissue was detected by the technique of immunofluorescence. The protein levels of p-AMPK/AMPK and apoptosis-related proteins were detected by Western blot. The expression of miR-33 in the liver tissues was detected by real-time PCR. RESULTS: Compared with model group, the contents of FBG, TG, TC, LDL-C, ALT and AST were decreased significantly, while the content of HDL-C was increased significantly in low-dose liraglutide group and high-dose liraglutide group (P<0.05). The protein levels of phosphorylated AMPK and Bcl-2 were up-regulated significantly, and the expression of cleaved caspase-3 was down-regulated significantly (P<0.05). The level of miR-33 was decreased significantly (P<0.01). CONCLUSION: Liraglutide alleviates liver injury in type 2 diabetic mice, and the mechanism may be associated with reducing the level of miR-33 and increasing the phosphorylation of AMPK in the liver tissues, thereby inhibiting hepatocyte apoptosis.     

杜仲叶提取物对人结直肠癌细胞HCT116及抗血管生成活性的影响

研究了杜仲叶提取物(EULE)对人结直肠癌细胞HCT116体外增殖和凋亡的影响以及其抗血管生成的活性。采用噻唑蓝(MTT)比色法测定了EULE及阳性对照药顺铂对HCT116和人脐静脉内皮细胞(HUVEC)体外增殖的影响;用流式细胞仪测定了EULE对HCT116细胞凋亡的影响。结果表明:EULE对HCT116细胞增殖具有一定的抑制作用,并表现出较好的量效关系,但抑制率稍低于顺铂;EULE对HCT116细胞的凋亡率的影响不显著;EULE对HUVEC细胞增殖的作用具有浓度依赖性,表现为低浓度促进增殖,高浓度抑制增殖。当EULE质量浓度分别为100、200、400和800 mg/L时,对HCT116细胞的抑制率分别为6.50%、10.52%、16.99%和56.49%,对HCT116细胞的凋亡率分别为0.00%、5.90%、7.00%和4.50%。EULE 800 mg/L时对HUVEC细胞的抑制率为41.59%,说明杜仲叶提取物只有达到一定浓度才具有抗新生血管生成的活性。     

HL-60 Cells Apoptosis Induced by Low-dose Cytosinearabinoside in Vitro

Cytosinearabinoside(Ara-C) is an important agent used for treatment of leukemia, but its mechanism of action at low dose is not clear, maybe is related to its effectson differentiationor apoptosis of cells. The authors investigate The relationship between bcl-2, p53 expression and apoptosis of HL-60 cells induced by low-dose cytosinearabinoside (10~(-8)M) by means of TUNEL method, flow cytometry, immunohistochemistry and in situ hybridization. The results show that low-doseAra-C could inhibit cell growth and induce apoptosis, and the effects might be related to increase of P53protein and decrease of bcl-2 mRNA expression.     

Reserarch advance on mechanism of arsenic trioxide on tumor

The article demonstrates the relative concepts of apoptosis and summarizes the effect of arsenic trioxide to the carcinoma, its possible principle included. Recent development of the reports about arsenic trioxide-induced apoptosis is referred and reviewed. Apoptosis is a natural mechanism and a common physiological death of cell, and its abnormity has great effect on cancer. Arsenic trioxide can induce apoptosis in APL cell and other cancer cells, and has been used in treating the patient with other kinds of malignant cancers. The mechanisms of arsenic trioxide-induced apoptosis are gradually becoming a hotspot, but still not lucid. With intensive study of arsenic trioxide, it can be a useful ways in treating cancer patients in the future.     

The toxic mixing zone of neutral and acidic river water: Acute aluminium toxicity in brown trout (Salmo trutta L.)

Mixing of acid river water containing aluminium (pH 5.1, Al 345 g.l^–1) with neutral water of a lake (pH 7.0, Al 73 g.l^–1) resulted in water (pH 6.4, Al 245 g.l^–1) with a pH (6.4) and Al concentration (245 g.l^–1) expected to have low toxicity to fish on the basis of current Al toxicity models. However, under semi-field conditions the freshly mixed water (a few sec. after mixing) proved to be highly toxic to brown trout. The fish were exposed to the water at different places along a 30 m channel. At the beginning of the channel acid and neutral water were continuously mixed; the mixed water left the channel after 340 sec. The cells of the gills showed a highly increased rate of cell death by apoptosis and necrosis. Intercellular spaces were enlarged, and many leucocytes penetrated in these spaces. Mucus release was stimulated to depletion. Plasma chloride levels were hardly affected. There was a clear gradient in the deleterious effects on the fish along the channel. The fish at the beginning of the channel (about 12 sec. after mixing of the water), were severely affected, whereas the fish kept at the end of the channel (340 sec. after mixing) were only mildly affected. In the natural situation fish will relatively quickly pass through a mixing zone. In our study we therefore focused on the effects on fish after a 60 min exposure to a mixing zone (5 sec after mixing), with subsequent recovery in a region downstream of the confluence and in neutral water with low Al. The recovery in the downstream area (at the end of the channel, i.e. 5 min after mixing) was clearly hampered when compared to the recovery in neutral water with low aluminium. Thus, a short exposure to the toxic mixing zone followed by a stay in water downstream of this zone, as may occur in nature, is detrimental to migrating trout. We conclude that freshly mixed acid and neutral water contain toxic components during the first seconds to minutes after mixing, that can not be explained by current models on aluminium toxicity.     

生长激素对体外培养猪COCs影响的研究

研究了生长激素对猪COCs体外成熟过程中卵丘细胞扩展、卵丘细胞凋亡、卵母成熟及孤雌激活后卵裂的影响。结果表明:生长激素(STH)能够促进卵丘细胞扩展,抑制卵丘细胞凋亡,对卵母细胞的成熟和激活后卵裂呈现双重效应。在猪COCs培养液中添加0.15μg/mL STH成熟率(73.83%±1.80%)和卵裂率(64.76%±2.84%)显著高于对照组(mNCSU-23+PMSG(10IU/mL)+hCG(10 IU/mL))及添加0.01、0.05、0.1、2μg/mL STH组(P<0.05),在本实验中为猪卵母细胞体外成熟的最佳条件。     

钙稳态失衡在氯化镧诱导MDCC-MSB1细胞凋亡中的作用

为探讨钙稳态失衡在LsCl3诱导MDCC-MSB1细胞凋亡中的作用,MDCC-MSB1细胞常规培养于RPMI1640培养液中,加入终浓度为0.5,1,1.5,2,2.5,3,3.5和4 mmol·L-1的LaCl3,继续培养24 h后,应用MTT法检测细胞增殖抑制率,DNA Ladder法和TUNEL法检测细胞凋亡,以Fura-2为荧光探针检测细胞内[Ca2+]i的变化.结果表明,在LaCl3浓度为0.5～4 mmol·L-1时,细胞的增殖抑制率增加,细胞凋亡数量和细胞内[Ca2+]i呈升高趋势,并呈剂量-效应关系.这表明LaCl3能抑制MDCC-MSB1细胞的增殖,并可能通过改变[Ca2+]i而诱导其发生凋亡.