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目的观察缺氧复氧对IEC-6肠上皮细胞线粒体膜电位的影响,探讨抗休克复方中药-紫芪方对IEC-6肠上皮细胞缺氧复氧损伤的保护作用机制。方法建立IEC-6细胞缺氧复氧损伤模型;采用血清药理学方法,在细胞培养液中加入不同给药剂量获取的“紫芪方”药物血清(分别以10、20g·kg-1分为1次给药和间隔1h、2次给药,DJ-1,SJ-2);参附药物血清(间隔1h,2次给药20g·kg-1SF)及生理氯化钠血清(S)。采用紫外分光光度分析法测定乳酸脱氢酶(LDH)漏出量,以及采用激光共聚焦显微镜测定线粒体膜电位(MMP)变化。结果IEC-6细胞缺氧复氧损伤后,可致细胞LDH的漏出量明显升高,线粒体膜电位明显降低。紫芪方药物血清明显减轻上述损伤,与对照组比较,SJ-2组制备的药物血清能明显减少IEC-6细胞LDH的漏出量(P<0.01);并具有保护IEC-6肠上皮细胞线粒体膜电位的作用(P<0.01)。结论缺氧复氧可导致IEC-6肠上皮细胞LDH漏出量升高和线粒体膜电位降低,“紫芪方”药物血清可减少细胞LDH漏出量及保护线粒体膜电位,从而发挥细胞的保护作用。
Objective To observe the effect of hypoxia and reoxygenation on the mitochondrial membrane potential of IEC-6 intestinal epithelial cells, and to explore the protective mechanism of anti-shock compound Chinese herbal medicine-Xiqi Fang on the hypoxia-reoxygenation injury of IEC-6 intestinal epithelial cells. Methods The anoxia and reoxygenation injury model of IEC-6 cells was established. Serum of pharmacological method was used to add “Zichuangfang” drug serum with different doses in cell culture medium (10 and 20 g·kg-1 respectively). 1 dose and interval 1h, 2 doses, DJ-1, SJ-2); paracetamol serum (interval 1h, twice 20g kg-1SF) and physiological sodium chloride serum (S). The amount of lactate dehydrogenase (LDH) leakage was measured by UV spectrophotometry, and the changes of mitochondrial membrane potential (MMP) were determined by laser confocal microscopy. RESULTS: After anoxia-reoxygenation injury in IEC-6 cells, the leakage of LDH in cells was significantly increased and the mitochondrial membrane potential was significantly reduced. Ziqinfang drug serum significantly reduced the above-mentioned damage, compared with the control group, the drug serum prepared from the SJ-2 group can significantly reduce the leakage of LDH in IEC-6 cells (P<0.01); and it has the protection of IEC-6 intestinal epithelial cell mitochondria The role of membrane potential (P <0.01). Conclusion Hypoxia and reoxygenation can lead to increased LDH leakage and mitochondrial membrane potential in intestinal epithelial cells of IEC-6. “Zijiaofang” drug serum can reduce cell LDH leakage and protect mitochondrial membrane potential, thus exerting cell protection.