[Regulatory Role of the AKT1/AMPK Pathway in Oxidative Stress of Lens Epithelial Cells in Cataract Patients].

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:40964109", "kind": "science-record", "revision": 2, "content_hash": "sha256:0216b2951f7b2d73bd011e1857dc5f60a10df643b8e7c2f04d6b3e03f023dabc", "created_at": "2026-11-01T20:31:03Z", "updated_at": "2026-11-01T21:35:47Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "[Regulatory Role of the AKT1/AMPK Pathway in Oxidative Stress of Lens Epithelial Cells in Cataract Patients].", "doi": "10.12182/20250560203", "pmid": "40964109", "authors": ["Yan Huang", "Bingjing Lyu", "Kunyuan Tang", "Ke Hu"], "journal": "Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition", "year": 2025, "abstract": "Objective: To elucidate the regulatory role and the underlying molecular mechanisms of serine/threonine protein kinase 1 (AKT1) in the lens epithelium of patients with age-related cataract (ARC).\nMethods: 1) Differentially expressed genes in ARC were screened using bioinformatics analysis (Genecard and GEO database GSE213546), and key genes were identified through functional enrichment analysis (KEGG and GO). 2) An oxidative stress model of lens epithelial cells was established by treating HLE-B3 cells with 200 μmol/L H2O2 for 24 h. RT-qPCR and Western blot were performed to assess AKT1 gene and protein expression changes. 3) Model cells were randomly divided into a si-NC group transfected with si-NC plasmids and 3 parallel si-AKT1 groups transfected with 3 types of si-AKT1 plasmids. A control + si-NC group was also set up, in which HLE-B3 cells not treated with H2O2 were transfected with si-NC empty plasmids. The AKT1 gene and protein expression levels in the si-NC and si-AKT1 groups were determined by RT-qPCR and Western blot. Two si-AKT1 parallel groups demonstrating the most significant changes in expression levels were selected for further experiments. The protein expression levels of AMPK and phosphorylated AMPK (p-AMPK) in the si-NC group, the two selected si-AKT1 parallel groups, and the control + si-NC group were determined by Western blot. A si-AKT1 parallel group demonstrating significant changes in p-AMPK/AMPK values was selected and treated with Acadesine (AICAR), an AMPK agonist, to verify the role of the AMPK pathway. Western blot was performed to determine Bcl-2 and Bax protein levels in the si-NC group, the control + si-NC group, and the si-AKT1 group before and after the administration of AICAR. Flow cytometry was performed to measure apoptosis and reactive oxygen species (ROS) levels, while ELISA kits were used to assess the levels of superoxide dismutase (SOD), malondialdehyde (MDA), and reduced glutathione (GSH).\nResults: 1) Through bioinformatics analysis, 78 differentially expressed genes were identified, with AKT1 significantly upregulated in ARC samples (P < 0.05) and enriched in the AMPK pathway. 2) Compared with cells not treated with H2O2, AKT1 mRNA and protein expression increased in the oxidative stress model cells. 3) The p-AMPK/AMPK ratio was higher in the si-NC group than that in the control + si-NC group. In contrast, AKT1 knockdown suppressed AMPK pathway activity, with all the si-AKT1 groups showing a significantly decreased p-AMPK/AMPK ratio compared to that of the si-NC group (P < 0.05). Compared with the control + si-NC group, the si-NC group exhibited elevated ROS and MDA levels, increased apoptosis rate, reduced SOD and GSH levels, downregulated Bcl-2, and upregulated Bax (all P < 0.05). Compared to those in the si-NC group, these indicators were improved in the si-AKT1 group (all P < 0.05). However, compared with the findings before AICAR treatment, these effects were antagonized after AICAR treatment in the si-AKT1 group (all P < 0.05).\nConclusion: The oxidative stress-related gene AKT1 may be a key pathogenic factor in cataract, and AKT1 induces oxidative stress and apoptosis in lens epithelial cells by modulating AMPK pathway activity.", "claims": [{"id": "claim-method", "statement": "使用GEO2R标准化数据（limma包）的normalizeBetweenArrays函数依据表达中位数对样本进行标准化处理。通过比较白内障组织和对照组的基因表达值，采用Bonferroni法调整P值，并将调整后的P值（adj.p.Value）<0.05且差异倍数logFC> 2 的基因定义为差异表达基因。", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:40964109", "locator": "sections[6] \"数据获取、标准化及差异表达分析\" paragraph 2", "quote": "使用GEO2R标准化数据（limma包）的normalizeBetweenArrays函数依据表达中位数对样本进行标准化处理。通过比较白内障组织和对照组的基因表达值，采用Bonferroni法调整P值，并将调整后的P值（adj.p.Value）<0.05且差异倍数logFC> 2 的基因定义为差异表达基因。"}], "state": "complete"}, {"id": "claim-data-access", "statement": "通过Genecard（https://www.genecards.org/Search/Keyword?queryString=fibrosis）检索oxidative stress and fibrosis相关基因。", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:40964109", "locator": "sections[6] \"数据获取、标准化及差异表达分析\" paragraph 2", "quote": "通过Genecard（https://www.genecards.org/Search/Keyword?queryString=fibrosis）检索oxidative stress and fibrosis相关基因。"}], "state": "complete"}, {"id": "claim-target-result", "statement": "氧化应激相关基因AKT1可能是白内障的关键致病基因，且AKT1通过影响AMPK通路活性诱导晶状体上皮细胞氧化应激和凋亡。", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:40964109", "locator": "sections[0] \"Abstract\" paragraph 4", "quote": "氧化应激相关基因AKT1可能是白内障的关键致病基因，且AKT1通过影响AMPK通路活性诱导晶状体上皮细胞氧化应激和凋亡。"}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "氧化应激相关基因AKT1可能是白内障的关键致病基因，且AKT1通过影响AMPK通路活性诱导晶状体上皮细胞氧化应激和凋亡。", "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:40964109"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. 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RT-qPCR and Western blot were performed to assess AK", "si-NC and si-AKT1 groups were determined by RT-qPCR and Western blot. Two si-AKT1 parallel group", "E-B3 cells with 200 μmol/L H2O2 for 24 h. RT-qPCR and Western blot were performed to assess AK", "-NC and si-AKT1 groups were determined by RT-qPCR and Western blot. Two si-AKT1 parallel group"], "order_basis": "paper_text_order:605", "id": "assay:1"}, {"role": "data_generation", "definition_id": "western_blot", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["s with 200 μmol/L H2O2 for 24 h. RT-qPCR and Western blot were performed to assess AKT1 gene and prote", "i-AKT1 groups were determined by RT-qPCR and Western blot. Two si-AKT1 parallel groups demonstrating t", "the control + si-NC group were determined by Western blot. A si-AKT1 parallel group demonstrating sign", "ist, to verify the role of the AMPK pathway. Western blot was performed to determine Bcl-2 and Bax pro"], "order_basis": "paper_text_order:616", "id": "assay:2"}, {"role": "data_generation", "definition_id": "transfection", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["lls were randomly divided into a si-NC group transfected with si-NC plasmids and 3 parallel si-AKT1 g", "si-NC plasmids and 3 parallel si-AKT1 groups transfected with 3 types of si-AKT1 plasmids. A control", "hich HLE-B3 cells not treated with H2O2 were transfected with si-NC empty plasmids. The AKT1 gene and", "the control + si-NC group. In contrast, AKT1 knockdown suppressed AMPK pathway activity, with all t"], "order_basis": "paper_text_order:752", "id": "assay:3"}, {"role": "data_generation", "definition_id": "flow_cytometry", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": ["efore and after the administration of AICAR. Flow cytometry was performed to measure apoptosis and react"], "order_basis": "paper_text_order:1808", "id": "assay:4"}, {"role": "data_generation", "definition_id": "apoptosis_assay", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["CAR. Flow cytometry was performed to measure apoptosis and reactive oxygen species (ROS) levels, wh", "bited elevated ROS and MDA levels, increased apoptosis rate, reduced SOD and GSH levels, downregula", "aract, and AKT1 induces oxidative stress and apoptosis in lens epithelial cells by modulating AMPK"], "order_basis": "paper_text_order:1848", "id": "assay:5"}, {"role": "data_generation", "definition_id": "elisa", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": ["reactive oxygen species (ROS) levels, while ELISA kits were used to assess the levels of super"], "order_basis": "paper_text_order:1907", "id": "assay:6"}]}