三七总皂甙对大鼠急性胰腺炎腺泡细胞钙超载的调节

目的 探讨三七总皂甙（PNS）对急性重症胰腺炎（SAP）大鼠胰腺腺泡细胞的保护作用及其机制.方法 30只SD大鼠分为假手术组（SO组）、急性重症胰腺炎（SAP）、三七总皂甙预处理组（PNS组）.采用经肠壁逆行胰胆管注射法建立大鼠急性重症胰腺炎模型,SO组进腹后仅翻动胰腺和十二指肠后关腹,不注入牛磺胆酸钠.其中PNS组模型建立后立即予以腹腔注射三七总皂甙（50 mg/mL血塞通注射液（0.1 mL/100 g）,假手术组（SO组）、急性重症胰腺炎（SAP）组建模后注射生理盐水（0.1 mL/100g）.建模4h后检测大鼠血清淀粉酶AMS、胰腺病理学变化了解胰腺炎的发病阶段.处死大鼠后取胰腺组织,采用钙荧光指示剂Fluo-3-AM作用腺泡细胞,在荧光显微镜下观察胰腺腺泡细胞荧光强度.使用流式细胞仪定量分析、计算出单个腺泡细胞内荧光强度（fluorescent intensity,FI）的平均值,荧光强度值可以表示细胞内钙离子的浓度.结果 PNS和SAP两组大鼠血清AMS水平均显著增高（P＜0.05）,SAP组大鼠血清AMS水平显著高过PNS组（P＜0.05）.PNS和SAP两组大鼠胰腺组织病理学评分水平均显著增高（P＜0.05）.PNS组大鼠胰腺组织病理学评分显著低于SAP组（P＜0.05）.通过流式细胞仪检测钙离子浓度提示：SO组、SAP组、PNS组三组胰腺细胞内Ca2＋]i的荧光强度具有统计学差异.SAP组和PNS组较SO组大鼠胰腺细胞内Ca2＋]i的荧光强度均有显著升高（P＜0.05）.与SAP组相比,PNS组大鼠胰腺细胞内Ca2＋]i的荧光强度均有显著降低（P＜0.05）.结论 SAP的发生与细胞钙超载有关,并且钙超载程度越高,SAP的病情越重.PNS可以减轻胰腺细胞内钙超载,可以减轻胰腺组织病理学改变。

Effect of Panax notoginseng Saponins on Calcium Overload of Acinar Cells in Rats with Acute Pancreatitis

Objective To demonstrate the protective effect and its mechanism of Panax notoginseng saponins （PNS） on severe acute pancreatitis （SAP） in rats of pancreatic acinar cell. Methods Thirty SD rats were randomly divided into three groups： sham operation group （group SO） , severe acute pancreatitis group （group SAP） , and PNS preconditioning group （group PNS） . There were 10 rats in each group. Rat SAP model was induced by retrograde injection of 5% taurocholic acid sodium to pancreatic duct. Group SO was just treated with pancreas and duodenum turning after abdominal closure, not with taurocholic acid sodium injection. The rats in group PNS were given by intraperitoneal injection of 0.1 mL/100g PNS 50 mg/mL （xuesetong injection 0.1 mL/100g） once the models were established. And rats in the other two groups were given 0.9% physiological saline. Rats serum AMS were detected at 4 hours after modeling, and pancreatic pathological changes were observed to understand the pathogenesis of acute pancreatitis. The pancreatic tissue was obtained and the fluorescence intensity （FI） of acinar cells was observed by calcium fluorescent tracer Fluo-5-AM using fluorescence microscope observation. The average value of single acinar cell FI was analyzed and calculated quantitatively using flow cytometry, whiCh could represent the concentration of calcium ion in cells. Results In groups PNS and SAP, rats serum AMS levels and pancreatic tissue pathology scores were significantly increased （P〈 0.05） , and those in group SAP rats were significantly higher than in group PNS rats （P 〈 0.05） . The calcium ion concentrations showed that there were statistical differences in pancreas FI of intracellular Ca2＋]i among three groups. Compared with group SO, the pancreas FI of intracellular Ca2＋]I in rats of groups SAP and PNS were significantly increased （P 〈 0.05） . And compared with group SAP, FI of Ca2＋]i in PNS group was significantly decreased （P 〈 0.05） . Conclusion The occurrence