青年科學(xué)工作者論壇2005年第1期

《衛生研究》--2005年第一期

補鉻對糖尿病大鼠骨骼肌 IRS-1 基因表達的影響

孫忠吳蘊棠車(chē)素萍王夏 1 郭剛
天津醫科大學(xué)公共衛生學(xué)院，天津 300070

摘要 : 目的研究補鉻對糖尿病大鼠骨骼肌代謝相關(guān)基因表達的影響。
方法對前期研究分離到的與補鉻有關(guān)的糖尿病大鼠差異顯示的基因片段進(jìn)行克隆、測序，并進(jìn)行同源性分析。根據待檢測的基因序列進(jìn)行引物設計，進(jìn)行 RT-PCR 檢測。用激光密度掃描儀進(jìn)行光密度掃描分析，以目的基因與參考基因的灰度比值反映其表達水平。結果經(jīng)過(guò)克隆、測序、同源性比較，差顯片段 Cr-5 的堿基序列與 IRS-1 同源性為 100% 。糖尿病對照組 IRS-1.mRNA 表達量（ 0.791±0.038 ）低于正常對照組 (0.892 ± 0.053 ， P <0.05) ，糖尿病補鉻組 IRS-1 mRNA 的表達量 (0.822±0.066) 與糖尿病對照組相比有所恢復，但尚未恢復到正常水平 ( P >0.05) 。結論補鉻對糖尿病大鼠骨骼肌 IRS-1mRNA 表達有上調趨勢， Cr-5 將作為候選基因有待于進(jìn)一步研究。

關(guān)鍵詞 胰島素受體底物 -1(IRS-1) 糖尿病大鼠 RT-PCR 基因克隆鉻

中圖分類(lèi)號： R587.1 O614.611 Q593 文獻標識碼： 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
基金項目：天津市高等學(xué)校科技發(fā)展基金項目（ No. 01-20804 ）
天津市自然科學(xué)基金項目（ No. 9903606611 ）
作者簡(jiǎn)介：孫忠，碩士，副教授
1 中國科學(xué)院微生物研究所

硒蛋白對糖尿病小鼠血糖、 Ca 2+ 轉運以及 NO 系統影響的實(shí)驗研究

羅軍張世聯(lián) 王薇
河北省疾病預防控制中心，石家莊 050021

摘要 : 目的 研究硒蛋白對糖尿病小鼠血糖、 Ca 2+ 轉運及 NO 系統的調控作用。
方法體重 20.3 ± 1.7 g 昆明種雄性小鼠，腹腔注射 200mg /kg 體重， 2% 的四氧嘧啶造糖尿病（ DM ）模型。實(shí)驗分六組：正常對照組（Ⅰ）、正常 + 硒蛋白組（ 100 μ g Se /kg 體重）（Ⅱ）、糖尿病對照組（Ⅲ）、 DM+ 硒蛋白低劑量組（ 100 μ g Se /kg 體重）（Ⅳ）、 DM+ 硒蛋白高劑量組（ 300 μ g Se /kg 體重）（Ⅴ）、 DM+ 亞硒酸鈉組（ 100 μ g Se /kg 體重）（Ⅵ）。結果 Ⅴ組血糖（ 20.4 ± 6.3 mmol/L ）明顯低于Ⅲ組（ 45.3 ± 3.3 mmol/L ）， p ＜ 0.05 ；腎臟 Ca 2+ -ATPase 活性，Ⅴ組（ 0.90 ± 0.5 μ Mpi.mg -1 蛋白 . 小時(shí) -1 ）明顯高于Ⅲ組（ 0.35 ± 0.1 μ Mpi.mg -1 蛋白 . 小時(shí) -1 ）， p ＜ 0.05 ； NOS 活性，Ⅴ組（ 25.0 ± 4.3 U/ml ）明顯低于Ⅲ組（ 35.2±4.4 U/ml ）， p ＜ 0.05 。結論補硒劑量為 300 μ g Se /kg 體重的硒蛋白能夠顯著(zhù)的降低糖尿病小鼠血糖、提高腎臟 Ca 2+ -ATPase 活性和降低血漿 NOS 活性。

關(guān)鍵詞 : 硒蛋白糖尿病血糖 Ca 2+ 轉運 NO 系統
中圖分類(lèi)號： R587.1 Q513 Q581 文獻標識碼：

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

基金項目：河北省科技廳資助項目（ No.01276192D ）
作者簡(jiǎn)介：羅軍，男，碩士，助教

麻痹性貝毒在廣州市售經(jīng)濟貝類(lèi)中污染狀況分析

聶利華 , 江天久 , 楊維東 , 劉潔生（ 1 ）
暨南大學(xué)生命科學(xué)技術(shù)學(xué)院 , 廣州 510632

摘要：目的 通過(guò)對廣州市黃沙海產(chǎn)品批發(fā)市場(chǎng) 7 種經(jīng)濟貝類(lèi)為期一年的麻痹性貝毒 (Paralytic Shellfish Poison, PSP) 污染狀況的調查分析，了解海產(chǎn)品食用的安全性。

方法：毒性測定按照 AOAC 小白鼠法進(jìn)行，成份分析利用高效液相色譜 (HPLC) ，安全性評價(jià)采用聯(lián)合國糧農組織 (FAO) 及我國漁政漁港監督管理局制定的貝類(lèi)安全食用標準 (4MU/g 肉 ) 。結果在調查的 84 份貝樣中，染毒貝整體毒力 ( 消化腺與肉的加權平均毒力 ) 低于 4MU/g 肉，毒素最高含量?jì)H為 1.84 MU/g 肉，所有貝類(lèi)均在安全食用范圍之內。染毒貝類(lèi)主要為櫛孔扇貝 ( Chlamys Mimachlamys nobilis ) 和嵌條扇貝（ Pecten albicans ）。毒素在 2 個(gè)種 9 份樣品的消化腺中檢出，某些樣品肌肉組織出現小白鼠毒性反應。一份樣品消化腺中毒素含量高達 14.52MU/g 肉，高效液相色譜（ HPLC ）分析表明，該樣品毒素成份主要為 B1 、 GTX2/3 、 GTX1/4 及 C 類(lèi)。結論廣州市市售鮮貝 PSP 含量和檢出率整體水平均較低；貝中消化腺毒素含量及檢出率明顯高于肌肉，個(gè)別腺體毒素含量超出標準。毒素分布存在季節性差異，春季毒素比較強，夏秋季檢出率高。因此有針對性的加強貝毒監測非常必要。

關(guān)鍵詞： 貝類(lèi) 麻痹性貝毒監測
中圖分類(lèi)號： TS207.3 R155.55 文獻標識碼： 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

基金項目：國家重點(diǎn)基礎研究發(fā)展規劃項目 (2001CB409709) 、廣東省科技計劃項目 ( 2003C 20412) 、廣州市科技計劃項目 (2002J1-C0011, 2001-Z-056-01) 資助
作者簡(jiǎn)介：聶利華，女，碩士；通訊聯(lián)系人：劉潔生（ tywd@jnu.edu.cn ）

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青年科學(xué)工作者論壇2005年第1期