Li Chang, Fan Rong, Li Dong-Bo, Zhou Wei, Huang Yu-E, Wang Bin
Department of Emergency Orthopaedic Surgery, Sanya Hospital of Traditional Chinese Medicine, Sanya 572000.
Department of Orthopaedic Surgery, Haikou Third Hospital, Haikou 571199, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2022 Nov;38(6):617-621. doi: 10.12047/j.cjap.6348.2022.112.
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. 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. 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 (< 0.05), but the levels of BUN, Cr and UP24 of rats in HBSP group were decreased (<0.05). Compared with HBSP group, there were no significant differences in the above indexes in EPO group (>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 (<0.05). Compared with control group, the levels of Agrin and MuSK in skeletal muscle pulled tissue were significantly decreased (<0.05), and those of HBSP group and EPO group were significantly increased compared with injured group (<0.05), but there was no significant difference between HBSP group and EPO group (>0.05). 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.
探讨促红细胞生成素衍生肽(又称螺旋B表面肽,HBSP)对急性骨骼肌拉伤大鼠肾脏及聚集蛋白(聚集素,Agrin)水平的保护作用。将40只SPF级SD雄性大鼠随机分为对照组、损伤组、HBSP组和促红细胞生成素(EPO)组,每组10只。除对照组外,其余各组均建立急性骨骼肌拉伤动物模型。造模成功后,HBSP组和EPO组大鼠腹腔注射60 μg/kg HBSP和5000 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-伊红染色观察肾脏组织和骨骼肌拉伤组织的病理形态。采用原位末端转移酶标记法(TUNEL)检测肾组织细胞凋亡率。采用蛋白质免疫印迹法(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。与对照组相比,损伤组大鼠肾功能指标血清肌酐(Cr)、尿素氮(BUN)和24小时尿蛋白(UP24)水平升高(P<0.05),而HBSP组大鼠BUN、Cr和UP24水平降低(P<0.05)。与HBSP组相比,EPO组上述指标差异无统计学意义(P>0.05)。对照组肌纤维结构完整,纤维束形态和结构正常,间质无红细胞和炎性细胞浸润,无纤维组织增生。损伤组肌肉组织排列稀疏、不规则,间质增宽,有大量炎性细胞和红细胞浸润。HBSP组和EPO组红细胞和炎性细胞减少,肌纤维横纵纹清晰。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和明显的基质增生,肾囊肿扩张,有空泡形成和明显的炎性浸润,HBSP组和EPO组炎性浸润减少,肾小球肥大和增生减轻。对照组、损伤组、HBSP组和EPO组肾细胞凋亡率分别为(4.05±0.51)%、(26.30±2.05)%、(14.28±1.62)%和(16.03±1.77)%,各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和MuSK水平显著降低(P<0.05);与损伤组相比,HBSP组和EPO组Agrin和MuSK水平显著升高(P<0.05),但HBSP组与EPO组之间差异无统计学意义(P>0.05)。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。