Li Yong-Yu, Lu Xin-Yuan, Li Xue-Jin, Li Yan-Na, Li Kun, Chen Chang-Jie
Department of Pathophysiology, Institute of Digestive Disease, Medical School of Tongji University, Shanghai, China.
J Gastroenterol Hepatol. 2009 Jan;24(1):155-65. doi: 10.1111/j.1440-1746.2008.05592.x.
To investigate the effects of serum and ascitic fluid from rats with acute pancreatitis (AP) on cellular free calcium concentration ([Ca(2+)]i) of isolated rat pancreatic acinar cells, and the intervention of pyrrolidine dithiocarbamate (PDTC) and tetrandrine (Tet) to cellular calcium overload in AP.
AP was induced in Sprague-Dawley rats with a retrograde pancreatic duct injection of 3% sodium deoxycholate, and confirmed by histopathological examination and amylase activity assay. The rat serum and ascitic fluid were collected at 1, 5 and 10 h after AP induction, and used as irritants on isolated rat pancreatic acinar cells. The effects on intracellular [Ca(2+)]i, and cell viability were examined. Then, the antagonistic effects of different concentrations of PDTC and Tet were assessed.
The irritation with AP serum and ascitic fluid reduced the survival rate of the isolated rat pancreatic acinar cells and increased the cellular [Ca(2+)]i significantly (P < 0.05). As AP induction course prolonged, the stimulation effect of the AP serum and ascitic fluid intensified. In the pretreated acinar cells with PDTC or Tet, the decreased cell vitality reverted. The elevation of [Ca(2+)]i in the acinar cells significantly ameliorated (significant, P < 0.05; very significant, P < 0.01).
The serum and ascitic fluid from AP rats drastically elevate the [Ca(2+)]i in isolated pancreatic acinar cells and decrease cell vitality, while the pretreatment of cells with PDTC and Tet offsets the calcium overload irritated by the AP serum and ascitic fluid and protects these isolated acinar cells.
研究急性胰腺炎（AP）大鼠血清和腹水对分离的大鼠胰腺腺泡细胞游离钙浓度（[Ca(2+)]i）的影响，以及吡咯烷二硫代氨基甲酸盐（PDTC）和粉防己碱（Tet）对AP细胞钙超载的干预作用。
采用逆行胰管注射3%脱氧胆酸钠诱导Sprague-Dawley大鼠发生AP，并通过组织病理学检查和淀粉酶活性测定进行确认。在AP诱导后1、5和10小时收集大鼠血清和腹水，用作分离的大鼠胰腺腺泡细胞的刺激物。检测对细胞内[Ca(2+)]i和细胞活力的影响。然后，评估不同浓度的PDTC和Tet的拮抗作用。
用AP血清和腹水刺激可降低分离的大鼠胰腺腺泡细胞的存活率，并显著提高细胞[Ca(2+)]i（P<0.05）。随着AP诱导过程的延长，AP血清和腹水的刺激作用增强。在预先用PDTC或Tet处理的腺泡细胞中，降低的细胞活力得以恢复。腺泡细胞中[Ca(2+)]i的升高明显改善（显著，P<0.05；非常显著，P<0.01）。
AP大鼠的血清和腹水可显著提高分离的胰腺腺泡细胞中的[Ca(2+)]i并降低细胞活力，而用PDTC和Tet预处理细胞可抵消AP血清和腹水刺激引起的钙超载并保护这些分离的腺泡细胞。
