分辨魔角旋转核磁共振波谱在大鼠慢性胰腺炎分级中的应用

目的 观察大鼠实验性慢性胰腺炎(CP)进展过程中胰腺组织代谢产物变化特征,探讨其在CP分级中的价值.方法 Wistar大鼠36只,经尾静脉注入二丁基二氯化物(dibutyltin dichloride,DBTC)溶液8 mg/kg体重制备CP模型.注射后即刻(对照组)及7、14、21、28、35 d分批取大鼠胰腺组织,小部分行病理检查,大部分液氮冷冻后置-80℃保存.应用高分辨魔角旋转核磁共振波谱(HRMAS NMR)法检测胰腺组织的代谢产物.以非正常结构、管状复合物、腺体萎缩、纤维化、水肿及炎细胞浸润程度等作为基本指标进行病理分级.结果 胰腺组织的病理损伤严重程度随造模时间的推移逐渐增加,造模后7、14 d大鼠胰腺改变为轻度CP,造模后21、28、35 d大鼠胰腺病理改变为重度CP.重度CP大鼠胰腺组织中甜菜碱(Bet)、胆碱(Cho)化合物含量较轻度CP组及对照组大鼠明显增多,而天冬氨酸盐(Asp)、乳酸(Lac)、脂肪酸(FA)、异亮氨酸/亮氨酸/缬氨酸(Ile/Leu/Val)的含量明显减少.轻度CP与对照组大鼠胰腺组织的代谢产物含量无明显差异.结论 应用HR-MAS NMR分析胰腺组织代谢产物有助于区分重度CP与轻度CP.

High-resolution magic angle spinning nuclear magnetic resonance spectroscopic in the grading of chronic pancreatitts in rats

Objective To observe the metabolic characteristics of an experimental model of chronic pancreatitis,and to investigate its role in the grading of chronic pancreatitis.Methods Thirty-six Wistar rats were injected with dibutyltin chloride (DBTC) solution (8mg/kg) via the tail vein to establish the experimental model of chronic pancreatitis.The 36 rats were divided into 6 groups with 6 rats in each group.On0,7,14,21,28,35 days after modeling,rats was sacrificed and pancreatic tissue of the rats was harvested,and a small part was used for pathologic study,the majority part was kept at-80℃ under liquid nitrogen freezing.Metabolites of pancreatic tissue were determined by high-resolution magic angle spinning nuclear magnetic resonance spectroscopic (HR-MAS NMRS).On the basis of the abnormal structure,tubular complexes,gland atrophy,fibrosis,edema and inflammatory cell infiltration,chronic pancreatitis was graded.Results Pathologic study showed the severity of chronic pancreatitis gradually increased with time after modeling.The 7th,14th day after modeling,the pancreatic change was mild chronic pancreatitis; the 21st,28th,35th day,the pancreatic change was changed into severe chronic pancreatitis.Principal component analysis of HR-MAS NMRS showed that the betaine (Bet) and choline ( Cho)-contained components were significantly increased in severe chronic pancreatitis; while aspartate (Asp),lactate (Lac),isoleucine/leucine/valine (I/L/V) and fatty acid (FA) were significantly reduced when compared with those in mild chronic pancreatitis and normal pancreatic tissue.There was no significant difference in the amount of metabolic characteristics between mild chronic pancreatitis and normal pancreatic tissue.Conclusions HRMAS NMRS was helpful in distinguishing the severe chronic pancreatitis from mild chronic pancreatitis.