【摘要】：目的:已有研究報道多巴胺在中樞和外周多個層面發(fā)揮對糖代謝的調控作用。胰島β細胞自身可以合成多巴胺,多巴胺和多巴胺D2受體激動劑對胰島有抑制胰島素分泌的作用。多潘立酮(DPD)作為經典的多巴胺D2受體拮抗劑,在臨床上應用較為廣泛。因為它有明確的促進胃動力和止吐作用,故而經常被用來治療糖尿病胃病。有少數研究(臨床試驗和動物實驗)闡述了應用多潘立酮對胰島素分泌的影響,然而研究結論并不統一。所以本實驗室就多潘立酮對胰島素分泌的影響進行了在體和離體實驗,并且嘗試研究了相關的作用機制,為多潘立酮的臨床應用提供更多依據,并在多巴胺和能量代謝系統關系的研究領域增加新的證據。方法:1.給c57小鼠腹腔注射多潘立酮溶液,測定其對腹腔注射葡萄糖耐量(IPGTT)及同步胰島素釋放的影響。2.處死小鼠后,迅速打開小鼠腹腔,將小腸撥至一側,暴露并剝離一段膽管,找到膽總管與十二指腸交匯處,以止血鉗夾閉壺腹部乳頭區(qū),將配置好的膠原酶Ⅴ從剝離好的一段膽總管打入胰腺,小心剝離胰腺,38℃水浴震蕩消化16 min,手工挑取小鼠胰島。用Dispase II消化得到小鼠胰島β細胞。3.將分得的胰島培養(yǎng)過夜后,用放射性免疫分析法檢測多潘立酮干預對小鼠胰島素分泌的影響。4.采用鈣離子成像技術檢測多潘立酮對胰島β細胞內鈣離子濃度的影響。5.胰島培養(yǎng)過夜后,用ELISA分析法檢測多潘立酮干預對細胞內環(huán)磷酸腺甘(c AMP)含量的影響。結果:1.單次腹腔注射(IP)多潘立酮(DPD 5mg/Kg)或DMSO溶劑后檢測IPGTT中血糖和血漿胰島素濃度,結果提示多潘立酮增強了葡萄糖刺激的胰島素釋放,并且在30分鐘時濃度較對照組顯著升高(n=6 P0.05)。多潘立酮組血糖值在多個時點(5-60分)較對照組有升高趨勢,但無統計學差異。2.在離體胰島素分泌實驗中,高糖(16.7m M)時多潘立酮對分離的胰島有促胰島素分泌作用,并且其促分泌作用隨著藥物濃度的增加而呈現逐漸加強趨勢。低糖(2.8m M)時多潘立酮沒有促胰島素分泌作用。因而,在除外在體實驗中其他機制參與的情況下,該實驗證實了即刻作用后多潘立酮可以增強葡萄糖刺激的胰島素分泌(GSIS)。3.鈣離子成像技術顯示,在16.7m M葡萄糖的基礎上,多潘立酮可進一步升高β細胞內的鈣離子濃度和鈣離子振蕩頻率。4.在高糖環(huán)境下(16.7m M)孵育小鼠胰島,較溶劑對照組,多潘立酮可顯著增加胰島β細胞內的c AMP含量,高糖+溶劑對照組(n=12)vs高糖+50μmol/L DPD組(n=12),P0.05。結論:1.腹腔注射多潘立酮促進了c57小鼠IPGTT 30分鐘時點的胰島素分泌。2.在離體實驗中,多潘立酮對c57小鼠胰島具有葡萄糖依賴性的促胰島素分泌作用。3.多潘立酮通過增加β細胞內鈣離子濃度促進胰島素分泌。4.多潘立酮增加了β細胞內c AMP濃度,可能參與了促胰島素分泌機制。
[Abstract]:Aim: dopamine has been reported to regulate glucose metabolism at central and peripheral levels. Islet 尾 cells can synthesize dopamine, dopamine and dopamine D2 receptor agonists to inhibit insulin secretion. As a classic dopamine D 2 receptor antagonist, domperidone (DPD) is widely used in clinic. It is often used to treat diabetic gastropathy because it promotes gastric motility and stops vomiting. A few studies (clinical and animal) have described the effects of domperidone on insulin secretion, but the findings are not uniform. So we have carried out both in vivo and in vitro experiments on the effect of domperidone on insulin secretion, and have tried to study the related mechanism, which provides more evidence for the clinical application of domperidone. New evidence has also been added to the field of research on the relationship between dopamine and energy metabolism systems. Methods: 1. The effects of domperidone solution on glucose tolerance (IPGTT) and simultaneous insulin release were measured in c 57 mice. After the mice were killed, the abdominal cavity of the mice was quickly opened, the small intestine was moved to one side, a segment of the bile duct was exposed and stripped, and the junction of the common bile duct and the duodenum was found, so that the papilla area of the ampulla could be closed by the hemostatic forceps. The configured collagenase 鈪，

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