Background:Osteoarthritis(OA)is a common degenerative joint disease character-ized by the progressive degradation of articular cartilage.Mitochondrial dysfunction and autophagy,including mitophagy,have been implicated...Background:Osteoarthritis(OA)is a common degenerative joint disease character-ized by the progressive degradation of articular cartilage.Mitochondrial dysfunction and autophagy,including mitophagy,have been implicated in OA pathogenesis.Long noncoding RNAs(lncRNA)are emerging as key regulators in various cellular pro-cesses,but their roles in OA,particularly in chondrocytes,remain poorly understood.This study explores the involvement of lncRNA-GCH1 in regulating mitophagy and its impact on chondrocyte function and cartilage degradation in OA.Methods:Primary chondrocytes were isolated from the cartilage tissues of OA pa-tients and healthy controls.lncRNA-GCH1 expression was assessed using RNA-seq,reverse transcription quantitative polymerase chain reaction,and RNA fluorescence in situ hybridization.Functional assays,including Cell Counting Kit-8(CCK-8),colony formation,flow cytometry,and Western blotting,were used to evaluate the effects of lncRNA-GCH1 knockdown on chondrocyte proliferation,apoptosis,cell cycle,and mi-tophagy.Mitochondrial function was assessed by measuring adenosine triphosphate production,reactive oxygen species levels,and mitochondrial membrane potential.In vivo,a murine OA model was used to examine the impact of lncRNA-GCH1 knock-down on cartilage degradation.Results:lncRNA-GCH1 was upregulated in OA chondrocytes and localized in the cy-toplasm.Knockdown of lncRNA-GCH1 enhanced cell proliferation and arrested cell cycle in G0/G1.It also suppressed mitophagy,improved mitochondrial function,and reduced matrix-degrading enzyme expression-effects that were reversed by rapa-mycin treatment.Meanwhile,lncRNA-GCH1 knockdown reduced PTEN-induced ki-nase 1(PINK1)aggregation and in vivo local inhibition of PINK1 diminished cartilage degradation.Conclusion:lncRNA-GCH1 regulates mitophagy in OA chondrocytes,influencing mi-tochondrial function and matrix degradation.Targeting lncRNA-GCH1 may offer a potential therapeutic approach for OA treatment.展开更多
Objective:To investigate the protective effects of gypenoside XVII(GP-17)against cisplatin-induced acute kidney injury and to elucidate whether its mechanism involves the activation of PINK1/Parkin-mediated mitophagy....Objective:To investigate the protective effects of gypenoside XVII(GP-17)against cisplatin-induced acute kidney injury and to elucidate whether its mechanism involves the activation of PINK1/Parkin-mediated mitophagy.Methods:Sprague-Dawley rats were randomly divided into four groups:control,cisplatin,cisplatin+GP-17,and GP-17 alone.Cisplatin was administered intraperitoneally at 20 mg/kg to induce acute kidney injury,while GP-17 was given orally at 40 mg/kg/day for 7 d.The levels of serum creatinine and blood urea nitrogen,superoxide dismutase activity,and malondialdehyde content were measured.Histopathological analysis and transmission electron microscopy were also performed to evaluate the effects of GP-17 on renal injury.Moreover,the expression of mitophagy-related proteins,including PINK1,Parkin,LC3,and p62,and the mRNA expression of inflammatory markers were determined by Western blot and quantitative RT-PCR assays.Furthermore,human renal tubular epithelial HK-2 cells were treated with cisplatin and GP-17,with or without PINK1 siRNA transfection.Cell viability,apoptosis,reactive oxygen species levels,mitochondrial membrane potential,and the protein expression associated with the PINK1/Parkin pathway were measured.Results:In rats with cisplatin-induced acute kidney injury,GP-17 significantly ameliorated cisplatin-induced elevations in serum creatinine and blood urea nitrogen,attenuated tubular damage and mitochondrial ultrastructural injury,and reduced oxidative stress by increasing superoxide dismutase activity and decreasing malondialdehyde content.GP-17 further upregulated the protein levels of PINK1,Parkin,and LC3-Ⅱ/Ⅰratio while promoting p62 degradation,indicating enhanced mitophagic flux.In HK-2 cells,GP-17(20μM)co-treatment markedly attenuated cisplatin-induced cytotoxicity,apoptosis,reactive oxygen species overproduction,and mitochondrial depolarization.However,all these protective effects of GP-17 were completely abolished upon PINK1 knockdown.Conclusions:GP-17 protects against cisplatin-induced nephrotoxicity by activating PINK1/Parkin-mediated mitophagy,which facilitates the clearance of damaged mitochondria,alleviates oxidative stress,and inhibits renal cell apoptosis.These findings identify GP-17 as a promising candidate for mitigating chemotherapy-induced acute kidney injury.展开更多
基金Sun Yat-sen Memorial Hospital Clinical Research 5010 Program,Grant/Award Number:SYS-5010-202403Natural Science Foundation of Guangdong Province,Grant/Award Number:2024A1515012811+1 种基金Sun Yat-sen Scientific Research Project,Grant/Award Number:YXQH202202Guangdong Provincial Key Research and Development Program,Grant/Award Number:2023B1111050003。
文摘Background:Osteoarthritis(OA)is a common degenerative joint disease character-ized by the progressive degradation of articular cartilage.Mitochondrial dysfunction and autophagy,including mitophagy,have been implicated in OA pathogenesis.Long noncoding RNAs(lncRNA)are emerging as key regulators in various cellular pro-cesses,but their roles in OA,particularly in chondrocytes,remain poorly understood.This study explores the involvement of lncRNA-GCH1 in regulating mitophagy and its impact on chondrocyte function and cartilage degradation in OA.Methods:Primary chondrocytes were isolated from the cartilage tissues of OA pa-tients and healthy controls.lncRNA-GCH1 expression was assessed using RNA-seq,reverse transcription quantitative polymerase chain reaction,and RNA fluorescence in situ hybridization.Functional assays,including Cell Counting Kit-8(CCK-8),colony formation,flow cytometry,and Western blotting,were used to evaluate the effects of lncRNA-GCH1 knockdown on chondrocyte proliferation,apoptosis,cell cycle,and mi-tophagy.Mitochondrial function was assessed by measuring adenosine triphosphate production,reactive oxygen species levels,and mitochondrial membrane potential.In vivo,a murine OA model was used to examine the impact of lncRNA-GCH1 knock-down on cartilage degradation.Results:lncRNA-GCH1 was upregulated in OA chondrocytes and localized in the cy-toplasm.Knockdown of lncRNA-GCH1 enhanced cell proliferation and arrested cell cycle in G0/G1.It also suppressed mitophagy,improved mitochondrial function,and reduced matrix-degrading enzyme expression-effects that were reversed by rapa-mycin treatment.Meanwhile,lncRNA-GCH1 knockdown reduced PTEN-induced ki-nase 1(PINK1)aggregation and in vivo local inhibition of PINK1 diminished cartilage degradation.Conclusion:lncRNA-GCH1 regulates mitophagy in OA chondrocytes,influencing mi-tochondrial function and matrix degradation.Targeting lncRNA-GCH1 may offer a potential therapeutic approach for OA treatment.
基金supported by grants from the Health Commission of Zigong High-Level Talent Development Project(WJW-GCCRC007).
文摘Objective:To investigate the protective effects of gypenoside XVII(GP-17)against cisplatin-induced acute kidney injury and to elucidate whether its mechanism involves the activation of PINK1/Parkin-mediated mitophagy.Methods:Sprague-Dawley rats were randomly divided into four groups:control,cisplatin,cisplatin+GP-17,and GP-17 alone.Cisplatin was administered intraperitoneally at 20 mg/kg to induce acute kidney injury,while GP-17 was given orally at 40 mg/kg/day for 7 d.The levels of serum creatinine and blood urea nitrogen,superoxide dismutase activity,and malondialdehyde content were measured.Histopathological analysis and transmission electron microscopy were also performed to evaluate the effects of GP-17 on renal injury.Moreover,the expression of mitophagy-related proteins,including PINK1,Parkin,LC3,and p62,and the mRNA expression of inflammatory markers were determined by Western blot and quantitative RT-PCR assays.Furthermore,human renal tubular epithelial HK-2 cells were treated with cisplatin and GP-17,with or without PINK1 siRNA transfection.Cell viability,apoptosis,reactive oxygen species levels,mitochondrial membrane potential,and the protein expression associated with the PINK1/Parkin pathway were measured.Results:In rats with cisplatin-induced acute kidney injury,GP-17 significantly ameliorated cisplatin-induced elevations in serum creatinine and blood urea nitrogen,attenuated tubular damage and mitochondrial ultrastructural injury,and reduced oxidative stress by increasing superoxide dismutase activity and decreasing malondialdehyde content.GP-17 further upregulated the protein levels of PINK1,Parkin,and LC3-Ⅱ/Ⅰratio while promoting p62 degradation,indicating enhanced mitophagic flux.In HK-2 cells,GP-17(20μM)co-treatment markedly attenuated cisplatin-induced cytotoxicity,apoptosis,reactive oxygen species overproduction,and mitochondrial depolarization.However,all these protective effects of GP-17 were completely abolished upon PINK1 knockdown.Conclusions:GP-17 protects against cisplatin-induced nephrotoxicity by activating PINK1/Parkin-mediated mitophagy,which facilitates the clearance of damaged mitochondria,alleviates oxidative stress,and inhibits renal cell apoptosis.These findings identify GP-17 as a promising candidate for mitigating chemotherapy-induced acute kidney injury.
