| Objective:To investigate the role of heme oxygenase-1(HO-1)-PTEN Induced Putative Kinase 1(PINK1)/Parkin in reducing acute kidney injury by electroacupuncture of endotoxemia acute kidney injury(E-AKI)and its relationship with mitochondrial fusion/fission.Methods:A total of 40 SPF male SD rats were gathered and randomly divided into 4 groups(n=10)by random number table method in this experiment: blank control group(group C),lipopolysaccharide group(LPS group),electroacupuncture + lipopolysaccharide group(EA+LPS group),and non-acupoints electroacupuncture + lipopolysaccharide group(SEA+LPS group).Endotoxemia model was established by intraperitoneal injection of LPS(10mg/kg).Group C was injected with equal volume of saline intraperitoneally,and the LPS group was intraperitoneally injected with LPS 10mg/kg.The EA+LPS group,30 minutes of electroacupuncture was performed continuously for 5 days intraperitoneal injection of LPS was conducted after the end day of electroacupuncture on the fifth day.Meanwhile,by selecting the bilateral Zusanli and Shenshu acupoints,rats were subjected to the stimulation intensity of dilatational waves with a frequency of 2/15 Hz to induce slight muscle tremor.The SEA+LPS group was treated the same as the EA+LPS group except for electrical stimulation at 0.5cm next to the acupoint.Once rats were killed 6h after LPS injection,femoral vein blood was obtained from four groups of rats for further detecting.The levels of serum creatinine(Cr)and urea nitrogen(BUN)by the enzyme activity kit.The method of enzyme-linked immunosorbent assay(ELISA)was adopted to detect the levels of serum neutrophil gelatinase-associated lipid transport protein(NGAL),interleukin-6(IL-6),tumor necrosis factor(TNF-α)and kidney injury molecule-1(KIM-1).Besides,kidney tissue was taken for HE staining and scored for kidney injury.Specifically,the expressions of HO-1,PINK1,Parkin,mitochondrial fusion protein 2(Mfn2),optic atrophy protein 1(OPA1)and mitochondrial dynamic-related protein 1(Drp1)were detected by Western Blot.Mitochondrial morphology in renal tubular epithelial cells was observed using transmission electron microscopy.Results:Compared with group C,the levels of Cr,BUN,KIM-1 and NGAL in femoral venous blood associated with kidney injury increased in LPS,EA+LPS and SEA+LPS groups(P<0.05);the levels of IL-6 and TNF-α indicators of inflammatory factors increased(P<0.05);the HSK score of renal tissue increased(P <0.05);the expressions of HO-1,PINK1,Parkin,and Drp1 in renal tissue were up-regulated,and the expressions of Mfn2 and OPA1 were down-regulated(P<0.05).Compared with the LPS group,the levels of Cr,BUN,KIM-1,and NGAL in femoral venous blood declined in the EA+LPS group(P<0.05);the levels of inflammatory factor indicators IL-6 and TNF-α reduced(P<0.05);the HSK score of kidney tissue decreased(P<0.05);the expressions of HO-1,PINK1,Parkin,Mfn2 and OPA1 in kidney tissue were upregulated and the expression of Drp1 was down-regulated(P<0.05).The difference was not statistically significant in the SEA+LPS group(P>0.05).It can be observed from transmission electron microscopy that mitochondria in renal tubular epithelial cells in group C were oval and rod-shaped,with mitochondrial cristae clearly visible,and the inner pleats of the plasma membrane were neatly arranged.In LPS and SEA+LPS groups,mitochondria in the renal tubular epithelial cells were spherical,vacuolated and swollen,and some of mitochondrial cristae and the inner pleats of the plasma membrane disappeared simultaneously.Mitochondrial damage in renal tubular epithelial cells in the EA+LPS group was milder than that in LPS and SEA+LPS groups,and some of the inner plasma membrane folds could be observed.Conclusion: Electroacupuncture at both Zusanli and Shenshu acupoints can regulate mitochondrial fusion/fission and reduce acute kidney injury in endotoxemia and the regulatory mechanism may be related to the HO-1/PINK1-Parkin signaling pathway. |
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