青年科学工作者论坛2005年第1期

《卫生研究》--2005年第一期

补铬对糖尿病大鼠骨骼肌 IRS-1 基因表达的影响

孙忠吴蕴棠车素萍王夏 1 郭刚
天津医科大学公共卫生学院，天津 300070

摘要 : 目的研究补铬对糖尿病大鼠骨骼肌代谢相关基因表达的影响。
方法对前期研究分离到的与补铬有关的糖尿病大鼠差异显示的基因片段进行克隆、测序，并进行同源性分析。根据待检测的基因序列进行引物设计，进行 RT-PCR 检测。用激光密度扫描仪进行光密度扫描分析，以目的基因与参考基因的灰度比值反映其表达水平。结果经过克隆、测序、同源性比较，差显片段 Cr-5 的碱基序列与 IRS-1 同源性为 100% 。糖尿病对照组 IRS-1.mRNA 表达量（ 0.791±0.038 ）低于正常对照组 (0.892 ± 0.053 ， P <0.05) ，糖尿病补铬组 IRS-1 mRNA 的表达量 (0.822±0.066) 与糖尿病对照组相比有所恢复，但尚未恢复到正常水平 ( P >0.05) 。结论补铬对糖尿病大鼠骨骼肌 IRS-1mRNA 表达有上调趋势， Cr-5 将作为候选基因有待于进一步研究。

关键词 胰岛素受体底物 -1(IRS-1) 糖尿病大鼠 RT-PCR 基因克隆铬

中图分类号： R587.1 O614.611 Q593 文献标识码： A

The effects of chromium supplementation on IRS-1 expression of skeletal muscle in diabetic rats

Sun Zhong, Wu Yuntang, Che Suping, Wang Xia, et al.

College of Public Health , Tianjin Medical University , Tianjin 300070, China

Abstract ： Objective To study the effects of chromium supplementation on related gene expression to metabolism of skeletal muscle in diabetic rats. Methods cDNA fragments from the our related research were cloned, sequenced and its comparalarity analysis has been done. The RT-PCR were made using the primers designed according to the sequence of cDNA. Results The identities between the sequence of Cr-5 and IRS-1 is 100%. The expression level of DM group(0.791±0.038) is obviously lower than those of normal group(0.892±0.053, P <0.05). And The expression level of DM+Cr group(0.822±0.066) have some improvement compare to those of DM group ( P >0.05). Conclusion There is a tendency that Chromium supplementation can increase the expression level of IRS-1 mRNA of skeletal muscle in diabetic rats. It will be studied as a candidate gene in future research.

Key words : IRS-1; Diabetic Rat; RT-PCR; Gene clone; Chromium
基金项目：天津市高等学校科技发展基金项目（ No. 01-20804 ）
天津市自然科学基金项目（ No. 9903606611 ）
作者简介：孙忠，硕士，副教授
1 中国科学院微生物研究所

硒蛋白对糖尿病小鼠血糖、 Ca 2+ 转运以及 NO 系统影响的实验研究

罗军张世联王薇
河北省疾病预防控制中心，石家庄 050021

摘要 : 目的 研究硒蛋白对糖尿病小鼠血糖、 Ca 2+ 转运及 NO 系统的调控作用。
方法体重 20.3 ± 1.7 g 昆明种雄性小鼠，腹腔注射 200mg /kg 体重， 2% 的四氧嘧啶造糖尿病（ DM ）模型。实验分六组：正常对照组（Ⅰ）、正常 + 硒蛋白组（ 100 μ g Se /kg 体重）（Ⅱ）、糖尿病对照组（Ⅲ）、 DM+ 硒蛋白低剂量组（ 100 μ g Se /kg 体重）（Ⅳ）、 DM+ 硒蛋白高剂量组（ 300 μ g Se /kg 体重）（Ⅴ）、 DM+ 亚硒酸钠组（ 100 μ g Se /kg 体重）（Ⅵ）。结果 Ⅴ组血糖（ 20.4 ± 6.3 mmol/L ）明显低于Ⅲ组（ 45.3 ± 3.3 mmol/L ）， p ＜ 0.05 ；肾脏 Ca 2+ -ATPase 活性，Ⅴ组（ 0.90 ± 0.5 μ Mpi.mg -1 蛋白 . 小时 -1 ）明显高于Ⅲ组（ 0.35 ± 0.1 μ Mpi.mg -1 蛋白 . 小时 -1 ）， p ＜ 0.05 ； NOS 活性，Ⅴ组（ 25.0 ± 4.3 U/ml ）明显低于Ⅲ组（ 35.2±4.4 U/ml ）， p ＜ 0.05 。结论补硒剂量为 300 μ g Se /kg 体重的硒蛋白能够显著的降低糖尿病小鼠血糖、提高肾脏 Ca 2+ -ATPase 活性和降低血浆 NOS 活性。

关键词 : 硒蛋白糖尿病血糖 Ca 2+ 转运 NO 系统
中图分类号： R587.1 Q513 Q581 文献标识码：

Effects of selenoproteinon blood glucose 、 Ca 2+ transfer and NO System in diabetic Mice

Luo Jun, Zhang Shilian, Wang Wei

Hebei Center of Prevent and Control Disease, Shijiazhuang 050021 ， China

Abstract: Objective To study the controling effect of selenoprotein on blood glucose 、 Ca 2+ transfer and NO system in diabetic mice. Methods Male Kunming mice of (20.3±1.7) g body weight were injected 200mg /kg body weight 、 2% alloxan in abdomen to make diabetic model （ DM ） ,and were randomly divided into six groups ： normal control group (Ⅰ) ， normal+ selenoprotein group(100μg Se /kg body weight) (Ⅱ), DM control group (Ⅲ) ， DM+ lower dose selenoprotein group(100μg Se /kg weight) (Ⅳ), DM+ higher dose selenoprotein group(300μg Se /kg body weight) (Ⅴ) ， DM+ Na 2 SeO 3 group(100μg Se /kg body weight) (Ⅵ). Results The level of blood glucose in Ⅴ group(20.4±6.3 mmol/L ) were significantly lower than Ⅲ group （ 45.3±3.3 mmol/L ） P ＜ 0.05.The activity of Ca 2+ -ATPase of kidney in Ⅴ group(0.90±0.5μMpi.mg -1 protein.hour -1 ) is significant ly higher than Ⅲ group （ 0.35±0.1μMpi.mg -1 protein.hour -1 ） P ＜ 0.05 ， and the activity of NOS in Ⅴ group （ 25.0±4.3 U/ml ） is significantly lower than Ⅲ group （ 35.2±4.4 U/ml ） P ＜ 0.05. Conclusion Selenoprotein that supplemented selenium doseis 300 μg Se /kg weight could significantly decrease blood glucose 、 increase the activities of Ca 2+ -ATPase on kidney and weaken the activities of plasma NOS in diabetic mice.

Key words: selenoprotein ， diabetes ， blood glucose ， Ca 2+ transfer ， NO system

基金项目：河北省科技厅资助项目（ No.01276192D ）
作者简介：罗军，男，硕士，助教

麻痹性贝毒在广州市售经济贝类中污染状况分析

聂利华 , 江天久 , 杨维东 , 刘洁生（ 1 ）
暨南大学生命科学技术学院 , 广州 510632

摘要：目的 通过对广州市黄沙海产品批发市场 7 种经济贝类为期一年的麻痹性贝毒 (Paralytic Shellfish Poison, PSP) 污染状况的调查分析，了解海产品食用的安全性。

方法：毒性测定按照 AOAC 小白鼠法进行，成份分析利用高效液相色谱 (HPLC) ，安全性评价采用联合国粮农组织 (FAO) 及我国渔政渔港监督管理局制定的贝类安全食用标准 (4MU/g 肉 ) 。结果在调查的 84 份贝样中，染毒贝整体毒力 ( 消化腺与肉的加权平均毒力 ) 低于 4MU/g 肉，毒素最高含量仅为 1.84 MU/g 肉，所有贝类均在安全食用范围之内。染毒贝类主要为栉孔扇贝 ( Chlamys Mimachlamys nobilis ) 和嵌条扇贝（ Pecten albicans ）。毒素在 2 个种 9 份样品的消化腺中检出，某些样品肌肉组织出现小白鼠毒性反应。一份样品消化腺中毒素含量高达 14.52MU/g 肉，高效液相色谱（ HPLC ）分析表明，该样品毒素成份主要为 B1 、 GTX2/3 、 GTX1/4 及 C 类。结论广州市市售鲜贝 PSP 含量和检出率整体水平均较低；贝中消化腺毒素含量及检出率明显高于肌肉，个别腺体毒素含量超出标准。毒素分布存在季节性差异，春季毒素比较强，夏秋季检出率高。因此有针对性的加强贝毒监测非常必要。

关键词： 贝类麻痹性贝毒监测
中图分类号： TS207.3 R155.55 文献标识码： A

Analysis of paralytic shellfish poison of bivalves in seafood market in Guangzhou

Nie Lihua, Jiang Tianqiu, Yang Weidong, Liu Jiesheng*

Department of Biotechology, Jinan University , Guangzhou 510632, China

Abstact: Objective The investigations of the paralytic shellfish poisoning (PSP) from Huangsha seafood market of Guangzhou was performed to assess the risk of PSP in bivalves. Methods The concentration and profiles of PSP toxins in bivalves were determined by mouse bioassay of AOAC and high-performance liquid chromatography (HPLC). The risk assessment of PSP in bivalves was conducted with FAO and Chinese Administration Organization of Fish Culture and Seaport. Results The content of PSP detected was lower than the safe standard (4MU/g meat) in all of the 84 samples, one of which had the highest toxicity with 1.84 MU/g

muscle. These results suggested that the bivalves in seafood market was safe to feed. It was 9 samples' gland in 2 species that be detected to have PSP in the bivalves being researched, the muscles had few PSP. The concentration of PSP in one sample's gland exceeded the threshold of FAO (4MU/g) with 14.52MU/g meat ,and the profiles of PSP in the gland were B1 、 GTX2/3 、 GTX1/4 and C according to HPLC. Conclusion These results suggested that both of the concentration and detection rate of PSP of bivalves in seafood market in Guangzhou were low as a whole, but the content and discovery rate of PSP were far higher in glands than in the muscles, and the PSP content in one gland exceeded the threshold of Standard. The levels of PSP contamination in shellfish was characteristic of season . The toxins level in shellfish were the maximum in spring, but the frequency of toxins detected in shellfish was higher in summer and autumn, s o the detection and risk assessment of PSP in bivalves from seafood market was essential in the future.

Key words: bivalves 、 paralytical shellfish poisoning 、 detection

基金项目：国家重点基础研究发展规划项目 (2001CB409709) 、广东省科技计划项目 ( 2003C 20412) 、广州市科技计划项目 (2002J1-C0011, 2001-Z-056-01) 资助
作者简介：聂利华，女，硕士；通讯联系人：刘洁生（ tywd@jnu.edu.cn ）