青蒿素诱导急性胰腺炎胰腺腺泡细胞凋亡的实验研究

Apoptosis of Pancreas Acinar Cell Induced by Artemisinin in Acute Pancreatitis

目的探讨青蒿素(artemisinin)对急性胰腺炎胰腺腺泡细胞凋亡的影响,并了解是否减轻急性胰腺炎严重程度。方法腹腔内注射雨蛙素(cerulein)诱导大鼠胰腺炎模型建立,设正常对照组、胰腺炎组及青蒿素组,通过原位末端标记(TUNEL)法检测各组胰腺组织中腺泡细胞的凋亡指数,并用分光光度法检测髓过氧化物酶(MPO)活性。采用两步酶消化法分离正常大鼠胰腺腺泡细胞,离体分组培养,采用丫碇橙(AO)染色及caspase-3活性的检测,评估青蒿素对胰腺腺泡细胞凋亡的影响,并检测各组培养液中的乳酸脱氢酶(LDH)及淀粉酶(AMS)活性,评估胰腺腺泡细胞酶的释放。结果在体实验中,青蒿素组与胰腺炎组比较,胰腺腺泡细胞凋亡指数明显增高(P<0.05),MPO活性明显减低(P<0.05)。体外实验中,青蒿素组胰腺腺泡细胞凋亡指数及caspase-3活性较胰腺炎组明显增高(P<0.05),培养液中的LDH和AMS活性也明显降低(P<0.05)。结论青蒿素可促进大鼠急性胰腺炎胰腺腺泡细胞凋亡,减少胰酶释放及中性粒细胞激活,可减轻急性胰腺炎严重程度。

Objective To explore the effect of ar.temisinin on the apoptosis of pancreas acinar cells in acute pancreatitis （AP）, and to study whether artemisinin can relieve the severity of AP. Methods （1） In vivo experiment： twenty one Wistar rats were divided into the following 3 groups randomly： the normal control group, the AP group and the artemisinin group. The model of AP was established by injecting cerulein into the peritoneal cavity of rat. After establishment of AP in the artemisinin group, artemisinin was injected into the peritoneal cavity. Meanwhile normal saline was injected into the peritoneal cavity of rats of the normal control group and the AP group. The apoptosis of pancreas acinar cell was detected by terminal deoxynucleotidyl transferase mediated nick end labeling （TUNEL）. The activity of myeloperoxidase was detected by absorption spectrometry. （2） In vitro experiment： the pancreas aclnar cells of normal rats were isolated through two-step enzyme digestion, and cultured. These acinar cells were divided into 3 groups： the normal control group, the AP group and the artemisinin group. Then, the cells of AP group were cocultured with cerulein, and those of the artemisinin group were cocultured with cerulein and artemisinin. The apoptosis of pancreas acinar cells were detected by AO dyeing and the measurement of the activity of caspase-3. And the activity of LDH and AMS in the culture medium of each group were measured. Results （1） In vivo ： the apoptosis index of the artemisinin group was sigificantly increased and the activity of myeloperoxidase was obviously decreased compared with the AP group （P〈0.05）. （2） In vitro： the apoptosis index and the activity of caspase-3 of the artemisinin group were significantly increased compared with the AP group （P〈0.05） ; the activities of LDH and AMS of the artemisinin group were more decreased than those of the AP group （P〈0.05）. Conclusion Artemisinin could contribute to the apoptosis of rat pancreas acinar cells, decrease the releasing of trypsogen, alleviate the activation of neutrophil and relieve the severity of AP.