| Objective: To investigate the effect and mechanism of Coenzyme Q10(CoQ10)on renal interstitial fibrosis in the rat model of Unilateral ureteral obstruction(UUO).In order to provide experimental basis for clinical treatment and delay of renal fibrosis.Methods: Male Sprague-Dawley rats were randomly divided into 5 groups after the establishment of UUO model: Sham group,UUO group,UUO+CoQ10 group,UUO+Nec-1(Necrostatin-1,RIP1 kinase inhibitor)group and UUO+GSK872(RIP3kinase inhibitor)group.The Sham group was given free left ureter.The UUO group was given free and ligated left ureter.The UUO+CoQ10 group was given CoQ10(20mg/kg/d)by gavage.The UUO+Nec-1 group was given Nec-1(2mg/kg/d)by gavage and GSK872(1 mg/kg/d)by intraperitoneal injection in the UUO+GSK872 group,and the rats were executed after 7 days.The Sham group was given equal amount of saline.Histological changes were observed by HE staining,cell microstructure changes were observed by electron microscopy,and renal interstitial fibrosis degree was observed by Masson staining.The interleukin-1β(IL-1β)and interleukin-18(IL-18)and other inflammatory factors,fibrillogenic factor-transforming growth factor β1(TGF-β1),apoptotic factor-cysteine protease-3(Cleaved Caspase-3),and regulatory factors related to necroinflammatory pathways,namely receptor-interacting protein 1-receptor expression of receptor-interacting protein 3-pseudokinase hybrid kinase structural domain protein(RIP1-RIP3-MLKL)were detected by Western blot.Results: 1.Histopathological changes in the UUO model were observed in the tubulointerstitium with HE staining,showing tubular necrosis,vacuole formation,tubular basement membrane thickening,inflammatory cell infiltration,and tubular atrophy,suggesting good modeling of the UUO model.2.Electron microscopy results show that the atrophied,vacuolar degenerated tubules and bundled collagen fibers in the renal interstitium deposition of renal tubules microvilli disappear,epithelial cells appear swollen,microbubble changes,mitochondria and endoplasmic reticulum dilate,and lysosomes increase.3.Analysis of Masson staining results using the quantitative scoring system showed that the degree of interstitial fibrosis was significantly increased in the UUO group(P< 0.01).Compared with the UUO group,it has significantly decreased in the UUO+CoQ10,UUO+Nec-1 and UUO+GSK872 groups(P<0.05).4.Western blot: Compared with the Sham group,the expression levels of IL-1β and IL-18 were significantly higher in the UUO group(P<0.01),and the expression levels of the above two proteins were significantly lower in the UUO+CoQ10,UUO+Nec-1and UUO+GSK872 groups than in the UUO group(P<0.05).TGF-β1 expression was significantly increased in UUO group(P<0.01),and the elevated protein expression levels were reduced with CoQ10,Nec-1 and GSK872(P<0.05).Expression of the apoptotic factor Cleaved Caspase-3 was significantly higher in the UUO group compared with the Sham group(P<0.01),and Cleaved Caspase-3 expression decreased in each of the three drug treatment groups(P<0.05).Upregulation of the necroinflammatory pathway-related regulatory factors RIP1-RIP3-MLKL was observed in the UUO group.The application of the CoQ10,Nec-1,and GSK872 reversed the elevation of these indicators.Conclusion:1.CoQ10 has an effect of improving the renal fibrosis of rat models caused by UUO,accompanied by a phenomenon of necrotizing inflammation reduction mediated by RIP1-RIP3-MLKL,from which CoQ10 may be attenuated by down-regulating the expression of RIP1,RIP3 or MLKL attenuated renal fibrosis.2.CoQ10 has anti-inflammatory and anti-apoptotic effects,which may open up a new therapeutic pathway for preventing or delaying renal failure in patients with CKD. |
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