《STZ造模小知識第五版》—李景風
2017年3月21日
模型代做
2017年3月21日

?鏈脲佐菌素(STZ)

?鏈脲佐菌素(STZ)


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數據參數(Specifications)

中文名稱 鏈脲菌素;鏈脲佐菌素;2-脫氧-2-[[(甲基亞硝基氨基)羰基]-氨基]-D-吡喃葡萄糖
英文名稱 STZ;Streptozocin;2-Desoxy-2-(3-methyl-3-nitrosoureido)-D-glucopyranose
CAS識別號 18883-66-4
MDL識別號 MFCD00006607
EINECS識別號 242-646-8
Beilstein 2060675
分子式 C8H15N3O7
分子量 1457.38
分子結構圖 ?
產品性狀 溶于水,溶于乙醇和乙醚。鏈脲佐菌素是一種氨基葡萄糖-亞硝基脲(glucosamine-nitrosourea),是一種DNA烷基化試劑,能通過GLUT2葡萄糖轉運蛋白(GLUT2 glucose transport protein)獨自進入細胞。對胰腺胰島內胰島素誘發的β-細胞具毒性。鏈脲佐菌素對GLUT2 陽性神經內分泌腫瘤細胞有毒性作用。熔點:115℃ (水解)。
質量標準 外觀Appearance white to light yellow power

純度Purity≥98.9% (HPLC)

水分water≤3%(K.F)

NMR?Conforms to Structure

水溶解試驗solubility in water50MG/ML一種含N-亞硝基的化合物,在胰腺胰島充當一氧化氮的供體;

誘導產生糖尿病動物模式的胰島素分泌細胞的死亡。高效DNA甲基化試劑,能誘發染色體斷裂。對表達

GLUT2葡萄糖轉運體(GLUT2 glucose transporter)的神經內分泌腫瘤細胞系具細胞毒性。醫學研究上

鏈脲佐菌素用于誘導Type 1、 Type 2糖尿病。

儲存條件 冷藏保存,Stored at ≤ 0 °C ?建議-20°C
包裝說明 進口分裝(滅菌裝),正品保證。
技術延伸

鏈脲佐菌素(Streptozotocin,STZ)對機體組織毒性相對較小,動物存活率高,是目前國內外使用較多的制備糖尿病動物模型的藥物。STZ對一定種屬動物的胰島β細胞有選擇性破壞作用,能誘發許多動物形成防糖尿病人患的動物模型,一般采用大鼠和小鼠,雄性鼠造模的成模率明顯高于雌性鼠(我們多次驗證)。Ⅰ型糖尿病與Ⅱ型糖尿病動物模型的制備與STZ注射的劑量大小有著直接關系:大劑量注射時,由于直接引起胰島β細胞的廣泛破壞,可造成Ⅰ型糖尿病模型;而注射較少量STZ時,由于只是破壞一部分胰島β細胞的功能,造成外周組織對胰島素不敏感,同時給予高熱量配方飼料喂養,兩者結合誘發病理、生理改變,制備接近于Ⅱ型糖尿病人患的動物模型。

STZ常用給藥量推薦(大小鼠)

動物的注射體重不同, STZ劑量選取也不同。STZ單位劑量高低和體重大小存在反比趨勢。

列舉(鼠/腹腔/體重為注射時體重):

170-210克體重大鼠制備Ⅰ型糖尿病模型,選取70~55Mg/Kg一次給藥(或分次注射比如35?Mg/KgX2次);

230-280克體重大鼠(已經過高熱量飼料喂養4周以上)制備Ⅱ型糖尿病模型,選取35~50Mg/Kg一次;

18-20克體重小鼠制備Ⅰ型糖尿病模型,選取150-220Mg/Kg一次給藥(或分次100MgX2或50Mg/KgX4次);

30-35克體重小鼠(已經過配方飼料喂養4周以上)制備Ⅱ型糖尿病模型,選取70-120Mg/Kg一次給藥;

給藥劑量的選取,是影響成模率的最大因素,這里僅僅給出一些參考范圍。

具體劑量的選取,希望大家通過預實驗的結果來確定。以上數據僅能得出不同重量動物與STZ給藥量上的關系,即體重越大的動物單位給藥量要適當降低,當然體重大的動物總的STZ耗量仍是高于體重小的。

對于更多體重級別的給藥量,特別是超重或者涉及到補藥的,可以聯系我們團隊,我們將會免費分享一些經驗數據供大家參考。我們也可以承接各種模型代做業務,特別是在大小鼠糖尿病模型制備上,我們成本壓縮做的比較好,也在業內已經形成良好口碑。

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STZ單次注射和多次注射之間的換算關系

舉例:每次給藥50mg/Kg,連續給三次,總劑量累加為150mg/Kg ,這種方法與150mg/Kg一次給藥相比,前者死亡率可能得到緩解,但STZ的對胰島β細胞的破壞力不如后者(一次給藥)來得徹底。換句話說,多次注射,可能減少死亡,但一次注射效果明顯,如果劑量選取科學,一次注射可以直接到位。建議根據動物品種和抵抗能力,來決定給藥次數。目前大部分用戶采用一次注射方法。

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STZ分次注射的周期和次數選擇

周期:大劑量STZ注射后,藥力發揮主要集中在注射后的3-72小時,所以如果采用分次注射,希望在2-72小時內延續第二次注射,目前大部分使用者以24小時為周期居多。

小劑量注射STZ后,如果劑量偏低(低于理想值),有可能在第二周血糖有緩慢提升跡象,但隨著峰值放緩且后延,也要有血糖先高后低、最終回落的心里準備。

次數::從1~7次都用人選用過。根據使用這情況大致排序:1次 3次 ?2次 ?5次 ?7次

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STZ緩沖劑制備

檸檬酸緩沖液的配制:檸 檬 酸(FW:210.14)?2.1g加入雙蒸水100mL中配成A液檸檬酸鈉(FW:294.10)2.94g加入雙蒸水100mL中配成B液

將A、B液按比例混合(按1:1或1:1.32),測定并調節ph=4.2-4.5,即是所需STZ的緩沖液buffer。

經驗分享:可以等比縮小,少量制備,盡量現用現配以防止氧化(PH漂移)。

緩沖液的常見問題

如果緩沖液配完后PH值不在范圍內怎么調節?

偏酸加檸檬酸鈉(或雙蒸水),偏堿加檸檬酸。早期文獻追求摩爾濃度,個人認為不需要。

緩沖液可否貯藏再用?

可以。用的時候再次測PH值,保證合乎范圍,盡可能現用現配,防止氧化(PH漂移)。

緩沖液是否需要過濾?

必須過濾。我們贈送的檸檬酸/檸檬酸鈉通常為AR級化學試劑,內有雜質和其他離子,會影響動物對STZ的吸收。特別是直接靜脈注射,嚴格要求過濾。《STZ造模小知識第四版-李景風》

緩沖液是否需要滅菌?

關于滅菌,目前沒有發現是否滅菌對實驗的影響,我們主張是,如果條件允許,采用靜脈注射的使用者最好滅菌(高溫120℃消毒20min),而腹腔注射的用戶可以不做滅菌處理。

能否使用其他溶液配置緩沖液?

可以但不推薦。例如用醋酸等溶液,也可以做出,但其PH值不好控制。 STZ只有在PH4.2-4.5的時候才具有最佳藥力活性。

STZ腹腔注射和尾部(靜脈)注射的區別

尾部靜脈注射,藥物利用率較高,同比腹腔注射,可減少藥量,缺點是操作起來不如腹腔注射方便。

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STZ注射操作

過夜禁食后稱重,按大鼠總重量比稱取STZ,放一干燥滅菌瓶內,外用錫紙包好,將檸檬酸緩沖液及裝STZ的瓶子置冰浴(此時未溶解STZ!),一起帶到動物房,如果注射操作技術不熟練,應兩組交替注射,建議分組溶解STZ(按照1%稀釋比例),比如10只或15只鼠/組。★確保STZ在溶解后,30分鐘內用完(且越快越好)!

經驗分享:推注的速度快,更容易形成高血糖;推注的速度慢,相對的危險性較低。常規操作中多要求快速注射。當然,STZ的劑量是決定血糖高低的主要因素。★ STZ容易潮解(水解)失活,?快速稱取后仍嚴格要求干燥,推薦用干燥鋁箔(或錫箔)紙或干燥的稱量紙。《STZ糖尿病造模小知識第五版-李景風》

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關于未達標補注STZ

模型不達標者,穩定后補注STZ(未達到標準的,劑量選取要根據實際情況判斷),也可以成模。建議讓血糖恢復正常后再常規劑量注射;?對于補藥劑量,我們可以分享更多經驗數據供參考,可聯系北京博愛港團隊。

STZ的保存方法

-20℃,干燥避光保存。本品易潮解,如需反復取量,應避免其觸水受潮,受觸潮30分鐘后失效,這和注射時要求其快速注射是一個道理,即STZ水溶液不穩定。

多次稱取STZ時的操作方法和注意事項

STZ易水解,如確實需要多次稱取,要嚴格按照避免受潮的原則操作和存放。

操作方法:操作環境、盛裝容器、分裝工具都必須保證干燥。快速分裝無需冰浴,分裝后用封口膜密封瓶口,用鋁箔紙(或錫箔,即避光)將瓶子包好,裝入防潮塑料貼袋中(雙層更可取),并-20冷藏,可長期保存。本品保質期為2年以上。再次啟用時,開啟瓶口前應先使瓶口通風干卻。

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STZ溶解時是否需要冰浴環境

關于這個問題,我在06年和同事做過分組對照比較.多次驗證過。初步結論:即溶即注,能在20分鐘內完成,STZ藥效可正常表達,在30分鐘后,STZ水溶液會隨著時間逐漸水解失活,而相對較低的冰浴環境,確實可以適當降低STZ水溶液狀態下的失活速度。我們建議,如果把動物分成小組(每組幾只)注射,每次溶解后能在20分鐘內完成注射,完全可以不用冰浴;如果覺得在這個時長內完成比較緊張,建議冰浴操作。《STZ造模小知識第五版-李景風》

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<博愛港-代做模型/指標檢測>業務推薦

可根據用戶實驗方案,博愛港團隊可承接代造常規動物模型和相關指標的檢測業務。糖尿病模型模型代造承接數量大,成本壓縮優勢明顯,如有需求可聯系我們。

<博愛港-糖尿病高脂高糖飼料>推薦

博愛港配方一:維持料65%+豬油10%+蔗糖20%+膽固醇2.5%+膽酸鈉1%+混調劑(雞蛋麻油花生)1.5%

博愛港配方二:維持料60%+豬油12%+糖10%+奶粉6%+雞蛋5%+油炸花生5%+鹽1% +麻油1%

以上兩種配方料為博愛港團隊為糖尿病大小鼠模型量身定做的專屬配方料,原料貨源均經過嚴格把關;兩種配方料均已經過市場多年、數以千計只次的模型驗證,可放心使用。常年備有現貨,歡迎咨詢購買。

也可根據用戶配方需求定做(非常規添加劑需自行另購,起做數量為30公斤,交貨期順延一周)

<血糖儀/試紙/注射器/濾膜濾器> 推薦

????使用人用血糖儀即可,動物血糖儀探測范圍高(30以上),實則意義不大。如果動物房有固然好,否則不建議另購。注射器推薦選擇直徑小(精確度高)的,便于推液卡量(根據體重差別推注液量不同);濾器選用針頭濾器比較方便(其他帶泵濾器也可以),濾膜孔徑要求0.22、0,45的都可以。

我們有代理各類品牌的血糖試紙血糖儀、注射器、濾器等耗材,常年大量批量備貨,壓縮成本讓利給用戶,可供廣大用戶選擇,血糖試紙我們推薦用羅氏的。

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<作者聲明>

以上材料為本人根據自身經驗和用戶數據總結得出,不承擔任何數據責任,多有不足,僅供參考,也歡迎各位同仁指正,真心希望能給初次造模的同學帶來幫助,一起為糖尿病科學研究做些貢獻。

未經本人授權,不得轉載。僅供科研人員內部參考使用。

我們會不斷提升自身能力,為大家奉獻品質卓越的產品、嚴謹專業的服務。

————北京博愛港(專業服務商) 李景風

 

 

 

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