目的探究维持性血液透析(maintenance hemodialysis,MHD)患者血清Beclin1和微管相关蛋白1轻链3-Ⅱ(microtubule associated protein 1 light chain 3-Ⅱ,LC3-Ⅱ)表达水平与血管钙化(vascular calcification,VC)的相关性。方法选取2017年...目的探究维持性血液透析(maintenance hemodialysis,MHD)患者血清Beclin1和微管相关蛋白1轻链3-Ⅱ(microtubule associated protein 1 light chain 3-Ⅱ,LC3-Ⅱ)表达水平与血管钙化(vascular calcification,VC)的相关性。方法选取2017年4月~2020年9月三河市燕郊人民医院收治的MHD患者145例作为研究对象,根据是否发生VC,将患者分为MHD未并发VC组(n=75)和MHD并发VC组(n=70)。根据VC评估结果将MHD并发VC患者分为轻度VC组(n=36)、中度VC组(n=24)、重度VC组(n=10);另选取同期在该院进行体检的健康人80例作为对照组。收集受试者一般资料,酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测受试者血清Beclin1和LC3-Ⅱ的水平,Pearson法分析MHD患者血清Beclin1和LC3-Ⅱ水平与VC相关指标的相关性,二元logistic回归分析MHD患者发生VC的影响因素。结果与对照组相比,MHD未并发VC组和MHD并发VC组血清肌酐(serum creatinine,SCr)(72.48±18.26μmol/L vs 685.30±192.42μmol/L,733.98±206.35μmol/L)、血磷(1.28±0.42mmol/L vs 2.08±0.71mmol/L。2.86±0.87mmol/L)、血钙(1.54±0.45mmol/L vs 2.46±0.62mmol/L,2.98±0.77mmol/L)、全段甲状旁腺激素(intact parathyroid hormone,iPTH)(58.10±17.36pg/ml vs 634.58±172.44pg/ml,769.48±195.02pg/ml)水平升高,差异有统计学意义(F=100.197~500.960,均P<0.05);血清血红蛋白(hemoglobin,Hb)(134.82±35.16g/L vs 112.78±32.85g/L,103.66±27.91g/L),Beclin1(8.09±2.16μg/L vs 5.65±1.43μg/L,2.56±0.73μg/L)和LC3-Ⅱ(45.16±5.15μg/L vs 36.31±3.42μg/L,27.47±2.76μg/L)水平降低,差异有统计学意义(F=18.748~372.522,均P<0.05);与MHD未并发VC组相比,MHD并发VC组血磷、血钙及iPTH水平升高(t=7.136~9.723,均P<0.05),血清Beclin1和LC3-Ⅱ水平降低(t=16.605,18.982,均P<0.05)。轻度VC组、中度VC组和重度VC组血清Beclin1(4.35±0.71μg/L,3.49±0.57μg/L和1.91±0.26μg/L)和LC3-Ⅱ(31.12±3.32μg/L,25.65±2.62μg/L vs 20.47±1.76μg/L)水平依次降低,差异均有统计学意义(F=366.298,296.025,均P<0.05)。MHD患者血清Beclin1水平与血磷、血钙及iPTH均呈负相关(r=-0.674,-0.682,-0.597,均P<0.05);血清LC3-Ⅱ水平与血磷、血钙及i PTH也呈负相关(r=-0.648,-0.703,-0.674,均P<0.05)。二元logistic回归分析发现,Beclin1和LC3-Ⅱ水平偏低是MHD患者发生VC的危险因素(P<0.05)。结论MHD并发VC患者血清Beclin1和LC3-Ⅱ水平降低,与VC严重程度有关,是MHD患者发生VC的危险因素,可作为预测MHD患者发生VC的潜在生物学标志物。展开更多
AIM: To evaluate the association of complement factor H(CFH) and microtubule-associated protein 1 light chain 3 beta(MAP1LC3B) gene polymorphisms with the risk of age-related macular degeneration(AMD) in a high-altitu...AIM: To evaluate the association of complement factor H(CFH) and microtubule-associated protein 1 light chain 3 beta(MAP1LC3B) gene polymorphisms with the risk of age-related macular degeneration(AMD) in a high-altitude population. METHODS: The study group consisted of 172 participants with symptoms of AMD who were examined and diagnosed between January 2019 and June 2020. The control group was composed of 120 healthy individuals. Each participant was required to provide two milliliters of peripheral blood for DNA extraction. Two single nucleotide polymorphisms(SNPs) of CFH(rs1061170 and rs800292) and two SNPs of MAP1LC3B(rs8044820 and rs9903) were genotyped. The genotypes and allele frequencies of the SNPs in the study and control groups were further compared using Chi-square and Fisher’s exact tests. RESULTS: In a high-altitude population, the nominally significant differences of rs800292 and rs9903’s genotype AG frequencies were observed in the AMD group(P=0.034 and 0.004, respectively). The frequencies of allele G of rs800292 and allele A of rs9903 were also significantly dif ferent in the AMD group compared to the control [(P=0.034, OR=0.70, 95%CI: 0.50-0.98) and(P=0.004, OR=1.60, 95%CI: 1.15-2.22), respectively]. No significant differences in the genotype distributions(P=0.16 and 0.40, respectively) and allele frequencies(P>0.05) of rs1061170 and rs8044820 were observed in the AMD group.CONCLUSION: Genotype AG of rs800292 may be a protective factor for AMD. Conversely, rs9903 seems to be a risk factor for AMD. Therefore, allele G of rs800292 may be a protective factor, and allele A of rs9903, a risk factor for AMD in Qinghai high-altitude population.展开更多
Endoplasmic reticulum stress and mitochondrial dysfunction play important roles in Parkinson s disease,but the regulato ry mechanism remains elusive.Prohibitin-2(PHB2)is a newly discove red autophagy receptor in the m...Endoplasmic reticulum stress and mitochondrial dysfunction play important roles in Parkinson s disease,but the regulato ry mechanism remains elusive.Prohibitin-2(PHB2)is a newly discove red autophagy receptor in the mitochondrial inner membrane,and its role in Parkinson’s disease remains unclear.Protein kinase R(PKR)-like endoplasmic reticulum kinase(PERK)is a factor that regulates cell fate during endoplasmic reticulum stress.Parkin is regulated by PERK and is a target of the unfolded protein response.It is unclear whether PERK regulates PHB2-mediated mitophagy thro ugh Parkin.In this study,we established a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced mouse model of Parkinson’s disease.We used adeno-associated virus to knockdown PHB2 expression.Our res ults showed that loss of dopaminergic neurons and motor deficits were aggravated in the MPTP-induced mouse model of Parkinson’s disease.Ove rexpression of PHB2 inhibited these abnormalities.We also established a 1-methyl-4-phenylpyridine(MPP+)-induced SH-SY5Y cell model of Parkinson’s disease.We found that ove rexpression of Parkin increased co-localization of PHB2 and microtubule-associated protein 1 light chain 3,and promoted mitophagy.In addition,MPP+regulated Parkin involvement in PHB2-mediated mitophagy through phosphorylation of PERK.These findings suggest that PHB2 participates in the development of Parkinson’s disease by intera cting with endoplasmic reticulum stress and Parkin.展开更多
Organisms produce high levels of reactive oxygen species(ROS)to kill pathogens or act as signaling molecules to induce immune responses;however,excessive ROS can result in cell death.To maintain ROS balance and cell s...Organisms produce high levels of reactive oxygen species(ROS)to kill pathogens or act as signaling molecules to induce immune responses;however,excessive ROS can result in cell death.To maintain ROS balance and cell survival,mitophagy selectively eliminates damaged mitochondria via mitophagy receptors in vertebrates.In marine invertebrates,however,mitophagy and its functions remain largely unknown.In the current study,Vibrio splendidus infection damaged mitochondrial morphology in coelomocytes and reduced mitochondrial membrane potential(ΔΨm)and mitophagosome formation.The colocalization of mitochondria and lysosomes further confirmed that lipopolysaccharide(LPS)treatment increased mitophagy flux.To explore the regulatory mechanism of mitophagy,we cloned Bcl2/adenovirus E1 B 19 kDa protein-interacting protein 3(BNIP3),a common mitophagy receptor,from sea cucumber Apostichopus japonicus(Aj BNIP3)and confirmed that Aj BNIP3 was significantly induced and accumulated in mitochondria after V.splendidus infection and LPS exposure.At the mitochondrial membrane,Aj BNIP3 interacts with microtubule-associated protein 1 light chain 3(LC3)on phagophore membranes to mediate mitophagy.After Aj BNIP3 interference,mitophagy flux decreased significantly.Furthermore,Aj BNIP3-mediated mitophagy was activated by ROS following the addition of exogenous hydrogen peroxide(H2 O2),ROS scavengers,and ROS inhibitors.Finally,inhibition of BNIP3-mediated mitophagy by Aj BNIP3 small interfering RNA(si RNA)or high concentrations of lactate increased apoptosis and decreased coelomocyte survival.These findings highlight the essential role of Aj BNIP3 in damaged mitochondrial degradation during mitophagy.This mitophagy activity is required for coelomocyte survival in A.japonicus against V.splendidus infection.展开更多
Previous studies have shown that Vibrio splendidus infection causes mitochondrial damage in Apostichopus japonicus coelomocytes,leading to the production of excessive reactive oxygen species(ROS)and irreversible apopt...Previous studies have shown that Vibrio splendidus infection causes mitochondrial damage in Apostichopus japonicus coelomocytes,leading to the production of excessive reactive oxygen species(ROS)and irreversible apoptotic cell death.Emerging evidence suggests that mitochondrial autophagy(mitophagy)is the most effective method for eliminating damaged mitochondria and ROS,with choline dehydrogenase(CHDH)identified as a novel mitophagy receptor that can recognize non-ubiquitin damage signals and microtubule-associated protein 1 light chain 3(LC3)in vertebrates.However,the functional role of CHDH in invertebrates is largely unknown.In this study,we observed a significant increase in the mRNA and protein expression levels of A.japonicus CHDH(AjCHDH)in response to V.splendidus infection and lipopolysaccharide(LPS)challenge,consistent with changes in mitophagy under the same conditions.Notably,AjCHDH was localized to the mitochondria rather than the cytosol following V.splendidus infection.Moreover,AjCHDH knockdown using si RNA transfection significantly reduced mitophagy levels,as observed through transmission electron microscopy and confocal microscopy.Further investigation into the molecular mechanisms underlying CHDH-regulated mitophagy showed that AjCHDH lacked an LC3-interacting region(LIR)for direct binding to LC3 but possessed a FB1 structural domain that binds to SQSTM1.The interaction between AjCHDH and SQSTM1 was further confirmed by immunoprecipitation analysis.Furthermore,laser confocal microscopy indicated that SQSTM1 and LC3 were recruited by AjCHDH in coelomocytes and HEK293T cells.In contrast,AjCHDH interference hindered SQSTM1 and LC3 recruitment to the mitochondria,a critical step in damaged mitochondrial degradation.Thus,AjCHDH interference led to a significant increase in both mitochondrial and intracellular ROS,followed by increased apoptosis and decreased coelomocyte survival.Collectively,these findings indicate that AjCHDH-mediated mitophagy plays a crucial role in coelomocyte survival in A.japonicus following V.splendidus infection.展开更多
文摘目的探究维持性血液透析(maintenance hemodialysis,MHD)患者血清Beclin1和微管相关蛋白1轻链3-Ⅱ(microtubule associated protein 1 light chain 3-Ⅱ,LC3-Ⅱ)表达水平与血管钙化(vascular calcification,VC)的相关性。方法选取2017年4月~2020年9月三河市燕郊人民医院收治的MHD患者145例作为研究对象,根据是否发生VC,将患者分为MHD未并发VC组(n=75)和MHD并发VC组(n=70)。根据VC评估结果将MHD并发VC患者分为轻度VC组(n=36)、中度VC组(n=24)、重度VC组(n=10);另选取同期在该院进行体检的健康人80例作为对照组。收集受试者一般资料,酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测受试者血清Beclin1和LC3-Ⅱ的水平,Pearson法分析MHD患者血清Beclin1和LC3-Ⅱ水平与VC相关指标的相关性,二元logistic回归分析MHD患者发生VC的影响因素。结果与对照组相比,MHD未并发VC组和MHD并发VC组血清肌酐(serum creatinine,SCr)(72.48±18.26μmol/L vs 685.30±192.42μmol/L,733.98±206.35μmol/L)、血磷(1.28±0.42mmol/L vs 2.08±0.71mmol/L。2.86±0.87mmol/L)、血钙(1.54±0.45mmol/L vs 2.46±0.62mmol/L,2.98±0.77mmol/L)、全段甲状旁腺激素(intact parathyroid hormone,iPTH)(58.10±17.36pg/ml vs 634.58±172.44pg/ml,769.48±195.02pg/ml)水平升高,差异有统计学意义(F=100.197~500.960,均P<0.05);血清血红蛋白(hemoglobin,Hb)(134.82±35.16g/L vs 112.78±32.85g/L,103.66±27.91g/L),Beclin1(8.09±2.16μg/L vs 5.65±1.43μg/L,2.56±0.73μg/L)和LC3-Ⅱ(45.16±5.15μg/L vs 36.31±3.42μg/L,27.47±2.76μg/L)水平降低,差异有统计学意义(F=18.748~372.522,均P<0.05);与MHD未并发VC组相比,MHD并发VC组血磷、血钙及iPTH水平升高(t=7.136~9.723,均P<0.05),血清Beclin1和LC3-Ⅱ水平降低(t=16.605,18.982,均P<0.05)。轻度VC组、中度VC组和重度VC组血清Beclin1(4.35±0.71μg/L,3.49±0.57μg/L和1.91±0.26μg/L)和LC3-Ⅱ(31.12±3.32μg/L,25.65±2.62μg/L vs 20.47±1.76μg/L)水平依次降低,差异均有统计学意义(F=366.298,296.025,均P<0.05)。MHD患者血清Beclin1水平与血磷、血钙及iPTH均呈负相关(r=-0.674,-0.682,-0.597,均P<0.05);血清LC3-Ⅱ水平与血磷、血钙及i PTH也呈负相关(r=-0.648,-0.703,-0.674,均P<0.05)。二元logistic回归分析发现,Beclin1和LC3-Ⅱ水平偏低是MHD患者发生VC的危险因素(P<0.05)。结论MHD并发VC患者血清Beclin1和LC3-Ⅱ水平降低,与VC严重程度有关,是MHD患者发生VC的危险因素,可作为预测MHD患者发生VC的潜在生物学标志物。
文摘AIM: To evaluate the association of complement factor H(CFH) and microtubule-associated protein 1 light chain 3 beta(MAP1LC3B) gene polymorphisms with the risk of age-related macular degeneration(AMD) in a high-altitude population. METHODS: The study group consisted of 172 participants with symptoms of AMD who were examined and diagnosed between January 2019 and June 2020. The control group was composed of 120 healthy individuals. Each participant was required to provide two milliliters of peripheral blood for DNA extraction. Two single nucleotide polymorphisms(SNPs) of CFH(rs1061170 and rs800292) and two SNPs of MAP1LC3B(rs8044820 and rs9903) were genotyped. The genotypes and allele frequencies of the SNPs in the study and control groups were further compared using Chi-square and Fisher’s exact tests. RESULTS: In a high-altitude population, the nominally significant differences of rs800292 and rs9903’s genotype AG frequencies were observed in the AMD group(P=0.034 and 0.004, respectively). The frequencies of allele G of rs800292 and allele A of rs9903 were also significantly dif ferent in the AMD group compared to the control [(P=0.034, OR=0.70, 95%CI: 0.50-0.98) and(P=0.004, OR=1.60, 95%CI: 1.15-2.22), respectively]. No significant differences in the genotype distributions(P=0.16 and 0.40, respectively) and allele frequencies(P>0.05) of rs1061170 and rs8044820 were observed in the AMD group.CONCLUSION: Genotype AG of rs800292 may be a protective factor for AMD. Conversely, rs9903 seems to be a risk factor for AMD. Therefore, allele G of rs800292 may be a protective factor, and allele A of rs9903, a risk factor for AMD in Qinghai high-altitude population.
基金supported by the Key Science and Technology Research of Henan Province,No.222102310351(to JW)Luoyang 2022 Medical and Health Guiding Science and Technology Plan Project,No.2022057Y(to JY)Henan Medical Science and Technology Research Program Province-Ministry Co-sponsorship,No.SBGJ202002099(to JY)。
文摘Endoplasmic reticulum stress and mitochondrial dysfunction play important roles in Parkinson s disease,but the regulato ry mechanism remains elusive.Prohibitin-2(PHB2)is a newly discove red autophagy receptor in the mitochondrial inner membrane,and its role in Parkinson’s disease remains unclear.Protein kinase R(PKR)-like endoplasmic reticulum kinase(PERK)is a factor that regulates cell fate during endoplasmic reticulum stress.Parkin is regulated by PERK and is a target of the unfolded protein response.It is unclear whether PERK regulates PHB2-mediated mitophagy thro ugh Parkin.In this study,we established a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced mouse model of Parkinson’s disease.We used adeno-associated virus to knockdown PHB2 expression.Our res ults showed that loss of dopaminergic neurons and motor deficits were aggravated in the MPTP-induced mouse model of Parkinson’s disease.Ove rexpression of PHB2 inhibited these abnormalities.We also established a 1-methyl-4-phenylpyridine(MPP+)-induced SH-SY5Y cell model of Parkinson’s disease.We found that ove rexpression of Parkin increased co-localization of PHB2 and microtubule-associated protein 1 light chain 3,and promoted mitophagy.In addition,MPP+regulated Parkin involvement in PHB2-mediated mitophagy through phosphorylation of PERK.These findings suggest that PHB2 participates in the development of Parkinson’s disease by intera cting with endoplasmic reticulum stress and Parkin.
基金supported by the National Natural Science Foundation of China(32073003,32102825)Natural Science Foundation of Zhejiang Province(LZ19C190001)+1 种基金Key Project from Science Technology Department of Zhejiang Province(2019R52016)K.C.Wong Magna Fund in Ningbo University。
文摘Organisms produce high levels of reactive oxygen species(ROS)to kill pathogens or act as signaling molecules to induce immune responses;however,excessive ROS can result in cell death.To maintain ROS balance and cell survival,mitophagy selectively eliminates damaged mitochondria via mitophagy receptors in vertebrates.In marine invertebrates,however,mitophagy and its functions remain largely unknown.In the current study,Vibrio splendidus infection damaged mitochondrial morphology in coelomocytes and reduced mitochondrial membrane potential(ΔΨm)and mitophagosome formation.The colocalization of mitochondria and lysosomes further confirmed that lipopolysaccharide(LPS)treatment increased mitophagy flux.To explore the regulatory mechanism of mitophagy,we cloned Bcl2/adenovirus E1 B 19 kDa protein-interacting protein 3(BNIP3),a common mitophagy receptor,from sea cucumber Apostichopus japonicus(Aj BNIP3)and confirmed that Aj BNIP3 was significantly induced and accumulated in mitochondria after V.splendidus infection and LPS exposure.At the mitochondrial membrane,Aj BNIP3 interacts with microtubule-associated protein 1 light chain 3(LC3)on phagophore membranes to mediate mitophagy.After Aj BNIP3 interference,mitophagy flux decreased significantly.Furthermore,Aj BNIP3-mediated mitophagy was activated by ROS following the addition of exogenous hydrogen peroxide(H2 O2),ROS scavengers,and ROS inhibitors.Finally,inhibition of BNIP3-mediated mitophagy by Aj BNIP3 small interfering RNA(si RNA)or high concentrations of lactate increased apoptosis and decreased coelomocyte survival.These findings highlight the essential role of Aj BNIP3 in damaged mitochondrial degradation during mitophagy.This mitophagy activity is required for coelomocyte survival in A.japonicus against V.splendidus infection.
基金supported by the National Natural Science Foundation of China (32102825)Natural Science Foundation of Zhejiang Province (LQ22C190003)。
文摘Previous studies have shown that Vibrio splendidus infection causes mitochondrial damage in Apostichopus japonicus coelomocytes,leading to the production of excessive reactive oxygen species(ROS)and irreversible apoptotic cell death.Emerging evidence suggests that mitochondrial autophagy(mitophagy)is the most effective method for eliminating damaged mitochondria and ROS,with choline dehydrogenase(CHDH)identified as a novel mitophagy receptor that can recognize non-ubiquitin damage signals and microtubule-associated protein 1 light chain 3(LC3)in vertebrates.However,the functional role of CHDH in invertebrates is largely unknown.In this study,we observed a significant increase in the mRNA and protein expression levels of A.japonicus CHDH(AjCHDH)in response to V.splendidus infection and lipopolysaccharide(LPS)challenge,consistent with changes in mitophagy under the same conditions.Notably,AjCHDH was localized to the mitochondria rather than the cytosol following V.splendidus infection.Moreover,AjCHDH knockdown using si RNA transfection significantly reduced mitophagy levels,as observed through transmission electron microscopy and confocal microscopy.Further investigation into the molecular mechanisms underlying CHDH-regulated mitophagy showed that AjCHDH lacked an LC3-interacting region(LIR)for direct binding to LC3 but possessed a FB1 structural domain that binds to SQSTM1.The interaction between AjCHDH and SQSTM1 was further confirmed by immunoprecipitation analysis.Furthermore,laser confocal microscopy indicated that SQSTM1 and LC3 were recruited by AjCHDH in coelomocytes and HEK293T cells.In contrast,AjCHDH interference hindered SQSTM1 and LC3 recruitment to the mitochondria,a critical step in damaged mitochondrial degradation.Thus,AjCHDH interference led to a significant increase in both mitochondrial and intracellular ROS,followed by increased apoptosis and decreased coelomocyte survival.Collectively,these findings indicate that AjCHDH-mediated mitophagy plays a crucial role in coelomocyte survival in A.japonicus following V.splendidus infection.
