Shenkang Injection Treating Chronic Renal Failure by Inhibiting Endoplasmic Reticulum Stress and Oxidative Stress
Objective:To investigate the effect of Shenkang Injection in treatment of chronic renal failure(CRF)and its effect on renal endoplasmic reticulum stress and oxidative stress.Methods:Ten rats were randomly selected from 50 SD rats as the control group,and the rest were replicated the CRF model by gavage of adenine(200 mg·kg-1).The successful rats were randomly divided into the model group,the low-dose(2 g·kg-1),the high-dose(4 g·kg-1)and the N-acetylcysteine(NAC,100 mg·kg-1)groups,and were treated with intraperitoneal injections of the corresponding drugs.The pathological morphology of the kidneys was observed with HE stai-ning,and the renal phosphorylates were detected with immunohistochemistry and Western blotting.The pathological morphology of kid-ney was observed with HE staining,and immunohistochemistry and Western blot were used to detect phosphorylated protein kinase R-like endoplasmic reticulum kinase(p-PERK),phosphorylated eukaryotic initiation factor-2alpha(p-ERK),phosphorylated eu-karyotic initiation factor-2alpha(p-ERK),and phosphorylated eukaryotic initiation factor-2alpha(p-ERK).eukaryotic initiation factor-2α(p-eIF2α),activating transcription factor 4(ATF-4),α-smooth muscle actin(α-SMA),and collagen I expression levels.Rat renal mesangial cells R5200 were treated with tert-butyl hydroperoxide(TBHP)to replicate the oxidative stress injury model.R5200 cells were divided into blank group,TBHP group(150 μmol·L-1 TBHP),TBHP+NAC group(150 μmol·L-1 TBHP+5 mmol·L-1 NAC),and TBHP+SK group(150 μmol·L-1 TBHP+50 mg·L-1 nephrokine injection-containing serum)and incu-bated for 24 h.The cells were treated with tert-butyl hydroperoxide(TBHP),and the model of oxidative stress injury was reproduced.DCFH-DA fluorescent probe was used to detect the level of reactive oxygen species(ROS),JC-1 staining was used to detect the lev-el of mitochondrial membrane potential(MMP),immunofluorescence was used to detect the level of ATF-4,and the expression levels of p-PERK,p-eIF2α and ATF-4 were detected with immunofluorescence and Western blot.Results:with HE staining,it was showed that there was no obvious abnormalities in the renal tubules of rats in the control group.Rats in the model group showed many renal tu-bular dilatation,hydropic degeneration of epithelial cells,and urate crystals and necrotic cells were seen in the renal tubules.The kid-neys of rats in the drug group showed a small amount of renal tubular dilatation,urate crystals and hydropic degeneration of epithelial cells.Immunohistochemistry showed that the expression levels of p-PERK,p-eIF2α,ATF-4,α-SMA,and Collagen I were in-creased in the kidneys of the rats in the model group compared with the control group(P＜0.01).Compared with the model group,the expression levels of p-PERK,p-eIF2α,ATF-4,α-SMA,Collagen I in the kidney of rats in the drug group were decreased(P＜0.01).With Western blot,it is showed that the expression levels of p-PERK,p-eIF2α in the kidney of rats in the model group were increased compared with the control group(P＜0.01).Compared with the model group,the expression levels of p-PERK and p-eIF2α in the kidneys of rats in the drug group were decreased(P＜0.05).Compared with that of the blank group,the cellular ROS level was increased and MMP level was decreased in TBHP group(P＜0.01).Compared with that of the TBHP group,cellular ROS level was decreased and MMP level was increased in TBHP+NAC group and TBHP+SK group(P＜0.01).With Western blot,it was showed that cellular p-PERK,p-eIF2α,ATF-4 expression level was increased in TBHP group compared with blank group(P＜0.01).Compared with that of the TBHP group,cellular p-PERK,p-eIF2α,ATF-4 expression levels decreased in TBHP+NAC group and TBHP+SK group(P＜0.05).Conclusion:Shenkang Injection can effectively alleviate CRF in rats,and its mechanism may be related to the inhibition of renal endoplasmic reticulum stress and oxidative stress response.
万方数据
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