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目的:制备并鉴定促红细胞生成素衍生肽(HBSP)并探讨其在大鼠肾脏缺血再灌注损伤中的保护作用及其机制。方法:制备并检测HBSP的纯度及分子量。200~220g SD大鼠随机分为假手术组、缺血组、HBSP组。阻断左侧肾蒂45min后切除右侧肾脏模型。术中及术后每6h给予HBSP,检测术后48h肾功能及炎症细胞因子肿瘤坏死因子-α(TNF-α),观察病理学改变,采用末端转移酶标记技术(TUNEL)法检测肾小管上皮细胞凋亡,并检测肾组织核转录因子-κB(NF-κB)活性。结果:HBSP纯度达98%,分子量与理论分子量吻合,HBSP组术后48h肾功能好于缺血组,组织中TNF-α水平及NF-κB活性显著低于缺血组(P<0.05),HE切片组织损伤减轻,TUNEL染色示HBSP组阳性细胞数显著少于缺血组(P<0.05)。结论:由促红细胞生成素成功制备线性多肽HBSP,其通过减轻炎性反应及抑制肾小管细胞凋亡,保护肾脏缺血再灌注损伤大鼠的肾功能。
OBJECTIVE: To prepare and identify erythropoietin-derived peptide (HBSP) and to explore its protective effect on renal ischemia-reperfusion injury in rats and its mechanism. Methods: The purity and molecular weight of HBSP were prepared and tested. 200 ~ 220g SD rats were randomly divided into sham operation group, ischemia group and HBSP group. Left renal pedicle block 45min after removal of the right kidney model. HBSP was given intraoperatively and postoperatively every 6h, renal function and inflammatory cytokines necrosis factor-α (TNF-α) were measured at 48h after operation. Pathological changes were observed. TUNEL method was used to detect renal tubule epithelium Apoptosis was detected and the activity of nuclear factor-κB (NF-κB) in renal tissue was detected. Results: The purity of HBSP was 98%. The molecular weight of HBSP was consistent with the theoretical molecular weight. The renal function of HBSP group at 48 hours after operation was better than that of ischemic group. The level of TNF-α and the activity of NF-κB in HBSP group were significantly lower than those in ischemic group (P <0.05) Tissue tissue injury was lessened in HE sections. TUNEL staining showed that the number of positive cells in HBSP group was significantly less than that in ischemia group (P <0.05). Conclusion: The linear peptide HBSP was successfully prepared by erythropoietin, which can protect renal function of renal ischemia-reperfusion injury rats by reducing inflammatory reaction and inhibiting apoptosis of renal tubular cells.