三七总皂苷对大鼠急性重症胰腺炎腺泡细胞钙超载的调节机制

Study on the Regulatory Mechanism of Panax Notoginseng Saponins on the Calcium Overload of Pancreatic Acinar Cells of Severe Acute PancreatitisRats

目的探讨三七总皂苷是否降低胰腺炎大鼠胰腺组织中钙调素依赖性蛋白激酶Ⅱ-γ(Ca MKⅡ-γ)mRNA表达,降低细胞钙超载的程度,减轻急性重症胰腺炎时胰腺组织的损伤。方法 15只SD大鼠随机分成假手术组(SO组)、急性重症胰腺炎组(SAP组)及三七总皂苷组(PNS组)。采用经肠壁逆行胰胆管注射法建立急性重症胰腺炎大鼠模型,假手术组开腹后仅翻动十二指肠。三七总皂苷组(PNS组)建模前1 h使用腹腔注射三七总皂苷(50 mg/m L血塞通注射液(0.1 m L/100 g)。所有实验分组在建模24 h后处死大鼠留取胰腺组织。使用流式细胞仪检测、分析腺泡细胞内钙离子浓度(使用荧光强度表示钙离子浓度),采用逆转录聚合酶链式反应检测胰腺组织CAMKⅡ-γmRNA表达的变化,并比较各组大鼠胰腺组织病理变化。结果 SAP组和PNS组胰腺组织中CAMKⅡ-γmRNA表达显著增高(P<0.05),PNS组较SAP组CAMKⅡ-γmRNA表达量显著下降(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+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+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+的荧光强度均有显著降低(P<0.05)。病理学评分:与SO组相比,PNS和SAP 2组大鼠胰腺组织水平均显著增高(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)。结论 SAP发生时,细胞钙超载程度越高,SAP的病情越重。CAMKⅡ-γ的表达量与胰腺炎病情轻重有关,三七总皂苷(PNS)干预可抑制胰腺细胞内CAMKⅡ-γ的表达,从而缓解胰腺细胞内钙超载,可以减轻胰腺组织病理学改变。

Objective： To investigate whether Panax Notoginseng Saponins （PNS） reduces pancreatic tissue pancreatitis calmodulin - dependent protein kinase Ⅱ - γ//（ CaMK Ⅱ - γ） mRNA expression of pancreas tissue in rats, the degree of intracellular calcium overload and the pancreas tissue damage of severe acute panereatitis. Methods ： 15 SD rats were randomly divided into a sham operation group （ SO group）, a severe acute pancreatitis group （SAP group） and a Panax notoginsenosides group （PNS group）. Severe acute pancreatitis rat models were made by intestinal wall retrograde cholangiopancreatography injection. The duodenums of SO rats were only flipped after their abdomens were opened. PNS group was given Panax saponins intraperitoneal injection （50 mg/mL and Xuesetong injection （0. 1 mL/100 g） 1 hour prior to modeling. All experimental rats were killed after modeling 24 hours later to take their pancreatic tissue. A flow cytometry was used to analyze the concentration of intracellular calcium and a reverse polymerase chain reaction used to determine the changes of CAMK ll - TmRNA. The pancreatic tissue pathological changes of each group were compared. Results ： The pancreatic tissue CAMK I1 - 3,mRNA expression of SAP group and PNS group significantly increased （ P 〈 0. 05 ） , but when compared with SAP group, CAMK I1 - ~/mRNA expression of PNS group significantly decreased （ P 〈 0. 05 ）. Through the detection of flow cytometry, calcium concentration indi- cated the pancreatic cell Ca^2 ＋ fluorescence intensity of the three groups had a statistically significant difference. The pancreatic cell Ca2＋ fluorescence intensity of SAP group and PNS significantly increased （ P 〈 0. 05 ） when compared with SO group, but when compared with SAP group, the pancreatic ceils Ca^2 ＋ fluorescence intensity of PNS group significantly decreased （ P 〈 0. 05 ）. Pathology score was when compared with SO group, the pancreatic tissue levels of PNS group and SAP group significantly increased （ P 〈 0.05 ）, but the pancreatic histopathological scores of PNS group were significantly lower than that of SAP group （ P 〈 0.05 ）. Conclusion ： When SAP occurs, higher the degree of intracellular calcium overload, more severe SAP＇ s illness is. CAMK Ⅱ - γexpression levels are related to the severity of pancreatitis and PNS intervention may inhibit the expression of pancreatic cells CAMK Ⅱ - γ, and thus relieve the pancreatic cell calcium overload and reduce pancreatic tissue pathological changes.