Zhu Jin Xia, Lo Pui Shan, Zhao Wen Chao, Tang Ning, Zhou Qing, Rowlands Dewi K, Gou Yu Lin, Chung Yiu Wa, Chan Hsiao Chang
Epithelial Cell Biology Research Center, Department of Physiology, The Chinese University of Hong Kong, Shatin, NT, PR China.
Cell Biol Int. 2002;26(12):1011-8. doi: 10.1006/cbir.2002.0960.
The present study examined the effect of Bak Foong Pills (BFP), an over-the-counter traditional Chinese medicine (China registration no. Z980035), on anion secretion and the underlying signaling pathways in normal and cystic fibrosis pancreatic duct cell lines, CAPAN-1 and CFPAC-1, respectively, using the short-circuit current technique. Apical addition of BFP ethanol extract (600 microg/ml) induced a fast transient I(SC) peak that was followed by a slower but more sustained increase in I(SC) in CAPAN-1 cells. However, the response to BFP in CFPAC-1 was predominantly the first transient peak. Apical addition of DIDS (200 microM) inhibited the first peak by more than 60% in both cell lines without significantly affecting the second I(SC) rise. More than 85% of the BFP-induced first transient in both cell lines was inhibited when extra and intracellular Ca(2+) was chelated or emptied by pre-treatment with BAPTA (100 microM) and thapsigargin (10 microM), respectively. Acute addition of PMA (1 microM), a PKC activator, blocked more than 95% of the BFP-induced first peak in both cell lines, consistent with previously reported PKC modulation of Ca(2+)-dependent pancreatic anion secretion. The BFP-induced second I(SC) rise in CAPAN-1 could be inhibited by 73.6% and 71.13% by pretreatment of the cells with MDL-12330A (20 microM), an adenylate cyclase inhibitor and Rp-cAMP (200 microM), a cyclic AMP antagonist, respectively. However, less than 25% of the I(SC) was inhibited by combined treatment with BAPTA and thapsigargin. The second rise was also completely blocked by DPC (2mM) or Glibenclamide (1mM). The results indicate that BFP ethanol extract stimulates pancreatic duct anion secretion in normal and CF cells via different signaling pathways involving both Ca(2+) and cAMP.
本研究采用短路电流技术，分别检测了非处方传统中药白凤丸（中国注册编号：Z980035）对正常胰腺导管细胞系CAPAN-1和囊性纤维化胰腺导管细胞系CFPAC-1阴离子分泌及相关信号通路的影响。在CAPAN-1细胞中，顶端添加白凤丸乙醇提取物（600微克/毫升）可诱导快速短暂的短路电流（I(SC)）峰值，随后I(SC)出现较慢但更持久的增加。然而，CFPAC-1细胞对白凤丸的反应主要是第一个短暂峰值。顶端添加二乙氨基二硫代甲酸钠（DIDS，200微摩尔）可使两种细胞系中的第一个峰值抑制超过60%，而对第二个I(SC)升高无显著影响。当分别用1,2-双（2-氨基苯氧基）乙烷-N,N,N',N'-四乙酸（BAPTA，100微摩尔）和毒胡萝卜素（10微摩尔）预处理细胞，螯合或排空细胞外和细胞内的钙离子时，两种细胞系中超过85%的白凤丸诱导的第一个短暂峰值受到抑制。急性添加蛋白激酶C（PKC）激活剂佛波酯（PMA，1微摩尔）可使两种细胞系中超过95%的白凤丸诱导的第一个峰值受到抑制，这与先前报道的PKC对钙离子依赖性胰腺阴离子分泌的调节作用一致。用腺苷酸环化酶抑制剂MDL-12330A（20微摩尔）和环磷酸腺苷（cAMP）拮抗剂Rp-cAMP（200微摩尔）预处理细胞，可分别使CAPAN-1细胞中白凤丸诱导的第二个I(SC)升高抑制73.6%和71.13%。然而，联合使用BAPTA和毒胡萝卜素处理时，I(SC)的抑制率不到25%。第二个升高也被二苯基胺氯磺酸盐（DPC，2毫摩尔）或格列本脲（1毫摩尔）完全阻断。结果表明，白凤丸乙醇提取物通过涉及钙离子和cAMP的不同信号通路刺激正常细胞和囊性纤维化细胞中的胰腺导管阴离子分泌。
