lopinavir  (MedChemExpress)

Journal: Bone Research

Article Title: AIDS patients suffer higher risk of advanced knee osteoarthritis progression due to lopinavir-induced Zmpste24 inhibition

doi: 10.1038/s41413-025-00431-2

Figure Lengend Snippet: Lopinavir strongly inhibits Col2a1 and Sox9 expression and accelerates chondrocyte senescence and degeneration in vitro. a , b RT-qPCR of Sox9 and Col2a1 after treatment with anti-HIV drugs; drugs in red boxes indicate >50% reduction. c RT-qPCR of Sox9 and Col2a1 under IL-1β and varying lopinavir concentrations. d , e Western blot and quantification of Sox9, Col2a1, and Mmp13 after IL-1β and lopinavir treatment. f , g Alcian Blue staining and quantification of micromass after IL-1β and lopinavir at different time points. h RT-qPCR of senescence-related genes ( p16 INK4a , p21 , p53 ) after IL-1β and lopinavir treatment. i Western blot and quantification of p16 INK4a and p21 in primary chondrocytes. j , k SA-β-gal staining and quantification in primary chondrocytes. Scale bar: 50 μm. l , m Cell cycle analysis after IL-1β and lopinavir treatment. n Heatmap of cell cycle and chondrogenesis-related gene expression after lopinavir treatment. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.000 1

Article Snippet: Lopinavir, ritonavir, and doxorubicin were purchased from MCE (New Jersey, United States) and dissolved in DMSO.

Techniques: Expressing, In Vitro, Quantitative RT-PCR, Western Blot, Staining, Cell Cycle Assay, Gene Expression

Journal: Bone Research

Article Title: AIDS patients suffer higher risk of advanced knee osteoarthritis progression due to lopinavir-induced Zmpste24 inhibition

doi: 10.1038/s41413-025-00431-2

Figure Lengend Snippet: Acceleration of osteoarthritis progression in mice by intraperitoneal injection of lopinavir. a Flowchart of animal experiment. b , c Representative images and quantitative analysis of 3D-reconstructed calcified tissues in different groups of mice at 4 weeks after DMM surgery, n = 5 or 6 per group. d Representative images of Safranine O staining, HE staining, and immunofluorescence staining for Sox9, Zmpste24, and p21 in different groups of mice 4 weeks after DMM surgery. Scale bar: 100 μm; e – i Quantitative analysis of OARSI score ( e ), synovial inflammation score ( g ), Safranine O-positive cartilage area ( f ), Sox9-positive cell ratio ( h ) and p21-positive cell ratio ( i ). Data are shown as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.000 1

Article Snippet: Lopinavir, ritonavir, and doxorubicin were purchased from MCE (New Jersey, United States) and dissolved in DMSO.

Techniques: Injection, Staining, Immunofluorescence

Journal: Bone Research

Article Title: AIDS patients suffer higher risk of advanced knee osteoarthritis progression due to lopinavir-induced Zmpste24 inhibition

doi: 10.1038/s41413-025-00431-2

Figure Lengend Snippet: The effect of lopinavir on cartilage aging and degeneration depends on Zmpste24 inhibition. a Representative images of Lamin A immunofluorescence staining in primary chondrocytes from wild-type mice and Zmpste24 -/- mice after lopinavir treatment. Scale bar: 10 μm. b Quantitative analysis of nuclear blebbing cells. c , d SA-β-gal staining and quantitative analysis in primary chondrocytes from wild-type mice and Zmpste24 -/- mice after lopinavir treatment. Scale bar: 100 μm. e RT-qPCR experiment to detect the mRNA expression levels of Zmpste24 , Col2a1 , Sox9 , and p21 in mouse primary cells after lopinavir treatment. f , g Toluidine Blue and Alcian Blue staining of micromass of different primary chondrocytes after lopinavir treatment, and quantitative analysis. h Western Blot analysis of Col2a1, Sox9, Lamin A, p16 INK4a , and p21 protein expression in different primary chondrocytes after lopinavir treatment. i Flowchart of the animal experiment. j Representative images of X-ray, micro-CT, HE staining, and Safranine O staining in wild-type mice and Zmpste24 -/- mice after further intraperitoneal injection of lopinavir/ritonavir following DMM surgery. Scale bar: 100 μm. n = 6, 7 or 8 per group. k – m Quantitative analysis of synovial inflammation score, Safranine O-positive cartilage area, and OARSI score. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.000 1

Article Snippet: Lopinavir, ritonavir, and doxorubicin were purchased from MCE (New Jersey, United States) and dissolved in DMSO.

Techniques: Inhibition, Immunofluorescence, Staining, Quantitative RT-PCR, Expressing, Western Blot, Micro-CT, Injection

Journal: Bone Research

Article Title: AIDS patients suffer higher risk of advanced knee osteoarthritis progression due to lopinavir-induced Zmpste24 inhibition

doi: 10.1038/s41413-025-00431-2

Figure Lengend Snippet: Zmpste24 deficiency induces senescence via Usp7/Mdm2/p53 pathway activation. a KEGG enrichment of differentially expressed genes after Zmpste24 inhibition by lopinavir. b Heatmap of p53 pathway-related gene expression. c RT-qPCR of p21 and Gadd45a after lopinavir treatment. d Immunofluorescence co-localization of Usp7 and Lamin A. Scale bar: 10 μm. e Co-immunoprecipitation of Lamin A and Usp7. f , g Western blot and quantification of Lamin A, Usp7, Mdm2, and p53 in wild-type and Zmpste24 -/- chondrocytes. h Immunofluorescence co-localization of Usp7 and Mdm2. Scale bar: 10 μm. i Immunoprecipitation and Western blot of Mdm2 binding to Usp7 after Zmpste24 knockdown. j , k Mdm2 degradation rate after cycloheximide treatment. l Mdm2 ubiquitination levels in primary chondrocytes. m , n Mdm2 immunohistochemical staining and quantification. Scale bar: 100 μm. n = 6/group. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.000 1

Article Snippet: Lopinavir, ritonavir, and doxorubicin were purchased from MCE (New Jersey, United States) and dissolved in DMSO.

Techniques: Activation Assay, Inhibition, Gene Expression, Quantitative RT-PCR, Immunofluorescence, Immunoprecipitation, Western Blot, Binding Assay, Knockdown, Ubiquitin Proteomics, Immunohistochemical staining, Staining

Journal: Bone Research

Article Title: AIDS patients suffer higher risk of advanced knee osteoarthritis progression due to lopinavir-induced Zmpste24 inhibition

doi: 10.1038/s41413-025-00431-2

Figure Lengend Snippet: Schematic diagram of this study. Patients in the PI group had lower KOOS scores and a higher incidence of radiological knee OA compared to the non-PI group. Mechanistically, lopinavir-induced Zmpste24 inhibition compromised nuclear membrane stability, reducing the interaction between the nuclear membrane-binding protein Usp7 and Mdm2. This disruption activated the Usp7/Mdm2/p53 pathway, accelerating cellular senescence

Article Snippet: Lopinavir, ritonavir, and doxorubicin were purchased from MCE (New Jersey, United States) and dissolved in DMSO.

Techniques: Inhibition, Membrane, Binding Assay, Disruption

Journal: Advanced Science

Article Title: The Compromised Fanconi Anemia Pathway in Prelamin A‐Expressing Cells Contributes to Replication Stress‐Induced Genomic Instability

doi: 10.1002/advs.202307751

Figure Lengend Snippet: Replication stress response defects and genomic instability in prelamin A‐expressing cells. a) SA‐β‐gal staining of IMR90‐hTERT fibroblasts treated with DMSO or LPV (20 µ m ) for 2 weeks. Scale bars: 100 µm. The percentage of SA‐β‐gal‐positive cells was calculated. n = 3. b) The level of γH2AX in prelamin A‐expressing ( ZMPSTE24 ‐knockdown or LPV‐treated) cells was measured by western blotting. n = 3. c) Immunofluorescence staining of 53BP1 in control and prelamin A‐expressing (LPV‐treated) cells with or without APH (0.4 µ m , 24 h) treatment. Scale bars: 10 µm. The number of 53BP1 bodies per nucleus was calculated. d) Immunofluorescence staining of lamin A/C in shCTRL and shZMPSTE24 cells. Scale bars: 10 µm. The percentage of micronuclei was calculated. n = 3. e) The proliferation of shCTRL and shZMPSTE24 cells was measured (left). The viability of shCTRL and shZMPSTE24 cells treated with the indicated concentrations of HU was measured (right). n = 3. f) Chromosomal aberrations, including breaks, gaps, rings, and dicentric chromosomes were analyzed for each metaphase in control and prelamin A‐expressing (LPV‐treated) cells with or without APH (0.3 µ m , 16 h) treatment. g) Chromosomal aberrations were analyzed for each metaphase in control and prelamin A‐expressing (LPV‐treated) cells with or without HU (4 mм, 4 h) and mirin (50 µ m , 4 h) treatment. Quantitative analysis results are shown as the mean ± SD. P values were determined by unpaired Student's t‐ test (a, b, d, and e) and the Mann‒Whitney test (c, f, and g). * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: LPV (MCE, HY‐14588), FTI‐277 (MCE, HY‐15872A), HU (Sigma–Aldrich, H8627), APH (Glpbio, GC10867), colcemid (Glpbio, GC40664), Nutlin‐3A (MCE, HY‐10029), and mirin (MCE, HY‐117693) were used.

Techniques: Expressing, Staining, Knockdown, Western Blot, Immunofluorescence, Control

Journal: Advanced Science

Article Title: The Compromised Fanconi Anemia Pathway in Prelamin A‐Expressing Cells Contributes to Replication Stress‐Induced Genomic Instability

doi: 10.1002/advs.202307751

Figure Lengend Snippet: Downregulation of the FA/BRCA gene network in prelamin A‐expressing cells. a) The expression of FANCD2 and RAD51 in ZMPSTE24 ‐knockdown cells was measured by western blotting. n = 3. b) mRNA levels of FA/BRCA genes in ZMPSTE24 ‐knockdown cells were measured by qPCR. n = 3. c) Western blotting was used to measure the expression of FANCD2 and RAD51 in cells treated with FTI (3 µ m , 48 h), LPV (20 µ m , 6 days) or LPV + FTI. The farnesylated prelamin A generated by LPV treatment (lane 3) migrated more rapidly than the nonfarnesylated prelamin A generated by FTI treatment (lane 2 and lane 4). n = 3. d) The expression of RAD51 in asynchronous, S‐phase‐synchronized and M‐phase‐synchronized control and ZMPSTE24 ‐knockdown cells was measured by western blotting. Asyn, asynchronous. n = 3. e) The expression of FANCD2 in asynchronous, G1‐phase‐synchronized, S‐phase‐synchronized and M‐phase‐synchronized control and ZMPSTE24 ‐knockdown cells was measured by western blotting. Asyn, asynchronous. n = 3. f) SA‐β‐gal staining of IMR90 fibroblasts at early (P6) and late (P20) passages. Scale bar: 100 µm. g) The protein levels of prelamin A, FANCD2, FANCI and RAD51 in IMR90 fibroblasts at early (P6) and late (P20) passages were detected by western blotting. Quantitative analysis results are shown as the mean ± SD. P values were determined by unpaired Student's t‐ test (a, b, d, and e) and one‐way ANOVA (c). ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: LPV (MCE, HY‐14588), FTI‐277 (MCE, HY‐15872A), HU (Sigma–Aldrich, H8627), APH (Glpbio, GC10867), colcemid (Glpbio, GC40664), Nutlin‐3A (MCE, HY‐10029), and mirin (MCE, HY‐117693) were used.

Techniques: Expressing, Knockdown, Western Blot, Generated, Control, Staining