[Intervention effects of erythropoietin derived peptide (HBSP) on renal injury induced by acute skeletal muscle strain in rats].

Chang Li, Rong Fan, Dong-Bo Li, Wei Zhou, Yu-E Huang, Bin Wang
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Abstract

Objective: To investigate the protective effects of erythropoietin derived peptide, also known as spiral B surface peptide (HBSP), on kidney and aggregated proteins (Agrin) levels in acute skeletal muscle strain rats. Methods: Forty SPF grade SD male rats were randomly divided into control group, injury group, HBSP group and EPO group, with 10 rats in each group. Acute skeletal muscle strain animal models were established except the control group. After successful modeling, the rats in HBSP group and EPO group were intraperitoneally injected with 60 μg/kg HBSP and 5 000 U/kg recombinant human erythropoietin (rhEPO), and the rats in the control group and the injured group were intraperitoneally injected with 0.9% normal saline. Renal function was monitored with relevant kits; Hematoxylin-eosin staining was used to observe the pathological morphology of kidney tissue and skeletal muscle strain tissue. The apoptosis rate of renal tissue cells was detected by in situ terminal transferase labeling (TUNEL). Western blot and quantitative polymerase chain reaction (Q-PCR) were used to determine the expressions of Agrin and muscular-specific kinase (MuSK) in the injured skeletal muscle of rats in each group. Results: Compared with the control group, the renal function indexes serum creatinine (Cr), urea nitrogen (BUN) and 24 h urinary protein (UP24) levels of rats in injured group were increased (P< 0.05), but the levels of BUN, Cr and UP24 of rats in HBSP group were decreased (P<0.05). Compared with HBSP group, there were no significant differences in the above indexes in EPO group (P>0.05). In the control group, the muscle fiber structure was intact, the shape and structure of the fiber bundles were normal, and there was no infiltration of red blood cells and inflammatory cells in the interstitium, and no fibrohyperplasia. In the injured group, the muscle tissue showed sparse and irregular arrangement, and the interstitial widened with a large number of inflammatory cells and red blood cell infiltration. Erythrocytes and inflammatory cells were reduced in HBSP group and EPO group, and the transverse and longitudinal lines of muscle were clear. The glomerular structure of the rats in the fibrohyperplasia control group was intact and no lesions were observed. In the injured group, glomerular hypertrophy and significant matrix hyperplasia were observed, as well as the expansion of renal cysts with vacuolar and significant inflammatory infiltration were observed, and the inflammatory infiltration was reduced in the HBSP and EPO groups. Glomerular hypertrophy and hyperplasia were alleviated. The apoptosis rates of kidney cells in control group, injured group, HBSP group and EPO group were (4.05±0.51) %, (26.30±2.05) %, (14.28±1.62) % and (16.03±1.77) %, respectively, and there were significant differences among these groups (P<0.05). Compared with control group, the levels of Agrin and MuSK in skeletal muscle pulled tissue were significantly decreased (P<0.05), and those of HBSP group and EPO group were significantly increased compared with injured group (P<0.05), but there was no significant difference between HBSP group and EPO group (P>0.05). Conclusion: Erythropoietin derived peptide (HBSP) has obvious intervention effects on renal function injury in rats with acute skeletal muscle strain, and its mechanisms may be related to reducing the apoptosis rate of renal tissue cells and activating Agrin and MuSK expression.

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[促红细胞生成素衍生肽(HBSP)对急性骨骼肌劳损大鼠肾损伤的干预作用]。
目的:探讨促红细胞生成素衍生肽(又称螺旋B表面肽(HBSP))对急性骨骼肌损伤大鼠肾脏和聚集蛋白(Agrin)水平的保护作用。方法:选取SPF级SD雄性大鼠40只,随机分为对照组、损伤组、HBSP组和EPO组,每组10只。除对照组外,其余动物均建立急性骨骼肌劳损模型。造模成功后,HBSP组和EPO组大鼠腹腔注射60 μg/kg HBSP和5 000 U/kg重组人促红细胞生成素(rhEPO),对照组和损伤组大鼠腹腔注射0.9%生理盐水。用相关试剂盒监测肾功能;采用苏木精-伊红染色法观察大鼠肾组织和骨骼肌劳损组织的病理形态。采用原位末端转移酶标记法(TUNEL)检测肾组织细胞凋亡率。采用Western blot和定量聚合酶链反应(Q-PCR)检测各组大鼠损伤骨骼肌中Agrin和肌肉特异性激酶(MuSK)的表达。结果:与对照组比较,损伤组大鼠肾功能指标血清肌酐(Cr)、尿素氮(BUN)、24 h尿蛋白(UP24)水平升高(P<0.05), HBSP组大鼠BUN、Cr、UP24水平降低(P<0.05)。与HBSP组比较,EPO组上述指标均无显著差异(P>0.05)。对照组肌纤维结构完整,纤维束形态结构正常,间质未见红细胞和炎性细胞浸润,未见纤维增生。损伤组肌肉组织稀疏,排列不规则,间质增宽,可见大量炎性细胞和红细胞浸润。HBSP组和EPO组红细胞和炎症细胞减少,肌肉横纵线清晰。纤维增生对照组大鼠肾小球结构完整,未见病变。损伤组可见肾小球肥大,明显基质增生,肾囊肿扩张伴空泡性浸润及明显炎性浸润,且HBSP组和EPO组炎症浸润均减轻。肾小球肥大和增生减轻。对照组、损伤组、HBSP组、EPO组肾细胞凋亡率分别为(4.05±0.51)%、(26.30±2.05)%、(14.28±1.62)%、(16.03±1.77)%,组间差异有统计学意义(P<0.05)。与对照组相比,骨骼肌拉伤组织中Agrin、MuSK水平显著降低(P<0.05), HBSP组和EPO组与损伤组相比显著升高(P<0.05),但HBSP组与EPO组之间差异不显著(P>0.05)。结论:促红细胞生成素衍生肽(HBSP)对急性骨骼肌拉伤大鼠肾功能损伤有明显干预作用,其机制可能与降低肾组织细胞凋亡率、激活Agrin和MuSK表达有关。
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