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ABSTRACT Purpose: The regulatory effect of microRNA on diseases has been confirmed. This study aimed to evaluate the expression of microRNA-210-3p in age-related cataracts and assess the effect of abnormal miR-210-3p expressions on H2O2-induced SAR01/04 cells. Methods: Reverse-transcription quantitative polymerase chain reaction method was performed to assess the levels of miR-210-3p in aqueous humor samples. Receiver operating characteristic analysis was employed to assess the discrimination ability of miR-210-3p between patients with age-related cataracts and healthy people, and Pearson correlation analysis was used to identify the correlation between miR-210-3p and oxidative stress indices such as superoxide dismutase, glutathione peroxidase, malonaldehyde. Cell counting kit-8 assay and Transwell assay were used to estimate the biological function of H2O2-induced age-related cataract cell model. The levels of oxidative stress indices such as superoxide dismutase, glutathione peroxidase, and malonaldehyde were measured to evaluate the degree of oxidative stress damage in the age-related cataract cell model. The relationship between miR-210-3p and its target gene was verified by luciferase reporter gene analysis. Results: The miR-210-3p expression was elevated in the aqueous humor of patients with age-related cataracts. A high miR-210-3p expression showed a high diagnostic value for age-related cataracts and was significantly associated with the level of oxidative stress markers in patients with age-related cataracts. The inhibition of miR-210-3p can reverse oxidative stress stimulation and adverse effects on H2O2-induced cell function. Conclusions: The results suggested that miR-210-3p could promote cell viability, cell migration, and oxidative stress by targeting autophagy-related gene 7 in in vitro age-related cataract cell model.
RESUMO Objetivo: O efeito regulador do microRNA em doenças tem sido confirmado, e este artigo tentou avaliar a expressão do microRNA-210-3p na catarata relacionada à idade e avaliar o efeito da expressão anormal do miR-210-3p em células SAR01/04 induzidas por H2O2. Métodos: O método de transcrição reversa seguida de reação em cadeia da polimerase (RT-PCR) quantitativa foi realizado para avaliar os níveis de miR-210-3p em amostras de humor aquoso. Análise de características operacionais do receptor foi feita para avaliar a capacidade de discriminação do miR-210-3p entre pacientes com catarata relacionada à idade e pessoas saudáveis. A análise de correlação de Pearson identificou a correlação do miR-210-3p e índices de estresse oxidativo, como superóxido dismutase, glutationa peroxidase, malonaldeído. O ensaio de contagem de células kit-8 (cck-8) e o ensaio no sistema Transwell foram utilizados para estimar a função biológica do formato de células de catarata relacionada com a idade induzida por H2O2. Os níveis de índices de estresse oxidativo como superóxido dismutase, glutationa peroxidase e malonaldeído foram detectados para avaliar o grau de dano do estresse oxidativo em formato de células de catarata relacionada à idade. A relação entre miR-210-3p e seu gene alvo foi verificada por análise do gene repórter luciferase. Resultados: A expressão miR-210-3p foi elevada no humor aquoso de pacientes com catarata relacionada à idade. A expressão miR-210-3p altamente expressiva mostrou alto valor diagnóstico para catarata relacionada à idade e foi significativamente associado ao nível de marcadores de estresse oxidativo em pacientes com catarata relacionada à idade. A inibição de miR-210-3p pode reverter a estimulação do estresse oxidativo e os efeitos adversos da função celular induzida por H2O2. Conclusões: Esses dados sugeriram que a expressão miR-210-3p poderia promover a viabilidade celular, migração celular e estresse oxidativo ao direcionar genes ATG 7 relacionados à autofagia em modelo in vitro de células de catarata relacionadas à idade.
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Objective To investigate the clinical significance of autophagy-related protein 7 (ATG7) in the diagnosis of HBV-related hepatocellular carcinoma (HBV-HCC) by measuring the expression level of serum ATG7 in patients with HBV-HCC. Methods A total of 50 patients with chronic hepatitis B (CHB) and 89 patients with HCC who were hospitalized in Mengchao Hepatobiliary Hospital of Fujian Medical University from June 2018 to December 2020 were enrolled, among whom 67 patients had HBV-HCC (HBV-HCC group) and 22 patients had no HBV-HCC (non-HBV-HCC group), and 20 healthy volunteers who underwent physical examination were enrolled as healthy control (HC) group. Demographic data and laboratory data including alpha-fetoprotein (AFP) were collected from each group, and ELISA was used to measure the serum level of ATG7. The receiver operating curve (ROC) was plotted for ATG7 and AFP used alone or in combination, and the area under the ROC curve (AUC) was compared. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups, and the Mann-Whitney U test was used for comparison between two groups; the chi-square test was used for comparison of categorical data between groups; a Spearman correlation analysis was used to investigate correlation. Results The serum level of ATG7 was 22.88(19.79-23.04) ng/mL in the HBV-HCC group, 17.06(14.45-19.40) ng/mL in the non-HBV-HCC group, 19.21(16.65-20.82) ng/mL in the CHB group, and 13.82(8.70-17.82) ng/mL in the HC group, with a significant difference between groups ( χ 2 =65.144, P < 0.001). ATG7 had an AUC of 0.818 (95% confidence interval [ CI ]: 0.743-0.879) and AFP had an AUC of 0.777 (95% CI : 0.698-0.843) in the diagnosis of HBV-HCC, suggesting that ATG7 had a slightly higher AUC than AFP ( Z =0.852, P =0.394). ATG7 combined with AFP had an AUC of 0.859 (95% CI : 0.790-0.913) in the diagnosis of HBV-HCC, which was significantly higher than the AUC of ATG7 alone ( Z =2.192, P =0.028) and AFP alone ( Z =2.076, P =0.038). Conclusion ATG7 is a good marker for the diagnosis of HBV-HCC, and combined measurement of ATG7 and AFP can significantly improve the diagnostic rate for HBV-HCC.
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Objective To investigate the clinical significance of autophagy-related protein 7 (ATG7) in the diagnosis of HBV-related hepatocellular carcinoma (HBV-HCC) by measuring the expression level of serum ATG7 in patients with HBV-HCC. Methods A total of 50 patients with chronic hepatitis B (CHB) and 89 patients with HCC who were hospitalized in Mengchao Hepatobiliary Hospital of Fujian Medical University from June 2018 to December 2020 were enrolled, among whom 67 patients had HBV-HCC (HBV-HCC group) and 22 patients had no HBV-HCC (non-HBV-HCC group), and 20 healthy volunteers who underwent physical examination were enrolled as healthy control (HC) group. Demographic data and laboratory data including alpha-fetoprotein (AFP) were collected from each group, and ELISA was used to measure the serum level of ATG7. The receiver operating curve (ROC) was plotted for ATG7 and AFP used alone or in combination, and the area under the ROC curve (AUC) was compared. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups, and the Mann-Whitney U test was used for comparison between two groups; the chi-square test was used for comparison of categorical data between groups; a Spearman correlation analysis was used to investigate correlation. Results The serum level of ATG7 was 22.88(19.79-23.04) ng/mL in the HBV-HCC group, 17.06(14.45-19.40) ng/mL in the non-HBV-HCC group, 19.21(16.65-20.82) ng/mL in the CHB group, and 13.82(8.70-17.82) ng/mL in the HC group, with a significant difference between groups ( χ 2 =65.144, P < 0.001). ATG7 had an AUC of 0.818 (95% confidence interval [ CI ]: 0.743-0.879) and AFP had an AUC of 0.777 (95% CI : 0.698-0.843) in the diagnosis of HBV-HCC, suggesting that ATG7 had a slightly higher AUC than AFP ( Z =0.852, P =0.394). ATG7 combined with AFP had an AUC of 0.859 (95% CI : 0.790-0.913) in the diagnosis of HBV-HCC, which was significantly higher than the AUC of ATG7 alone ( Z =2.192, P =0.028) and AFP alone ( Z =2.076, P =0.038). Conclusion ATG7 is a good marker for the diagnosis of HBV-HCC, and combined measurement of ATG7 and AFP can significantly improve the diagnostic rate for HBV-HCC.
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Aberrant activation of NLRP3 inflammasome in colonic macrophages strongly associates with the occurrence and progression of ulcerative colitis. Although targeting NLRP3 inflammasome has been considered to be a potential therapy, the underlying mechanism through which pathway the intestinal inflammation is modulated remains controversial. By focusing on the flavonoid lonicerin, one of the most abundant constituents existed in a long historical anti-inflammatory and anti-infectious herb
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Neural stem cells (NSCs) have the ability to self-renew and differentiate into neurons, oligodendrocytes, and astrocytes. Highly dynamic nature of NSC differentiation requires the intimate involvement of catabolic processes such as autophagy. Autophagy is a major intracellular degradation pathway necessary for cellular homeostasis and remodeling. Autophagy is important for mammalian development and its role in neurogenesis has recently drawn much attention. However, little is known about how autophagy is associated with differentiation of NSCs into other neural lineages. Here, we report that autophagy plays a critical role in differentiation of adult rat hippocampal neural stem (HCN) cells into astrocytes. During differentiation, autophagy flux peaked at early time points, and remained high. Pharmacological or genetic suppression of autophagy by stable knockdown of Atg7, LC3 or CRISPR-Cas9-mediated knockout (KO) of p62 impaired astrogenesis, while reintroduction of p62 recovered astrogenesis in p62 KO HCN cells. Taken together, our findings suggest that autophagy plays a key role in astrogenesis in adult NSCs.
Sujets)
Adulte , Animaux , Humains , Rats , Cellules souches adultes , Astrocytes , Autophagie , Différenciation cellulaire , Homéostasie , Cellules souches neurales , Neurogenèse , Neurones , Oligodendroglie , Suppression génétiqueRésumé
Objective To detect the expression of the autophagy gene Atg5 and Beclin-1 in cervical cancer.Methods Streptavidin peroxidase (SP) immunohistochemical method was used to detect the expression of Atg5 and Beclin-1 in 50 paraffin specimens of cervical cancer,50 CIN (cervical intraepithelial neoplasia) and 50 normal cervical tissues.Results (1) The positive rates of Atg5 and Beclin-1 were significantly lower in cervical cancer than that in normal cervical tissue(34％ vs 72％,26％ vs 96％),with statistically significant differences (P ≤ 0.05).(2) The expressions of Atg5 were correlated with clinical stage,tumor type and degree of differentiation of cervical cancer,with statistically significant differences (x2 =14.90,4.36,14.78,P ≤0.05).The expressions of Beclin-1 were not correlated with age,clinical stage,size,the type,pathological type of the tumors,degree of differentiation,the lymph nodes metastasis of cervical cancer (P ＞ 0.05).(3) There was a correlation between Atg5 and Beclin-1 protein expression in cervical cancer tissues (x2 =17.92,P ≤ 0.05).Conclusions The downregulation of Atg5 and Beclin-1 protein in cervical cancer may be related to the progression of cervical lesions.Atg5 is correlated with clinical stage,tumor type and cell differentiation of cervical cancer patients,suggesting that Atg5 can be used as a biological indicator to predict the occurrence and development of cervical cancer and to evaluate the prognosis of patients.The expression of Atg5 and Beclin-1 in cervical cancer tissues is correlated,suggesting that the two proeins may be involved in occurrence and development of cervical cancer.
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Objective:To determine mechanisms for the role of high mobility group box-1 (HMGB1) to platinum based chemo-sensitivity in cervical cancer.Methods:Using Western blot,we examined the effect of cisplatin on expressions of LC3,Beclin1 and P62 in cervical cancer.After treated with autophagy inhibitor and/or cisplatin in HeLa and CaSki cells,cell counting kit-8 (CCK-8) assay was used to evaluate the chemo-sensitivity.The expression of LC3,Beclin1 and P62 were detected by Western blot.We established HeLashHMGB1,CaSki-shHMGB1 and HeLa-CTR,CaSki-CTR stable cell lines.CCK-8 assay was used to determine the chemosensitivity in these cell lines.Using Western blot,we examined the correlation among the expressions of HMGB1,LC3,Beclin1,and P62.Results:With the increase of cisplatin,the expression of LC3 and Beclin1 was increased,while the expression of P62 was decreased in HeLa and CaSki cell lines.Combination of cisplatin with autophagy inhibitor could increase the cellular sensitivity to cisplatin (P＜0.05).The expression of HMGB1 was related to chemo-sensitivity and autophagy related protein in cervical cancer cells.HMGB1 was positively correlated with LC3 and Beclin1 while negatively correlated with P62 (P＜0.05).Conclusion:HMGB 1 might play an important role in chemo-sensitivity via regulating autophagic activity.The combination of cisplatin with autophagy inhibitor might become a possible strategy for cervical cancer.HMGB1 might serve as a potential biomarker for predicting chemo-therapeutic responses.
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Aim To observe the effect of capsaicin on the experimental autoimmune neuritis (EAN) in rats and explore the mechanism.Methods To induce EAN,male Lewis rats were immunized with peripheralnerve myelin sheath antigen (P257481) peptide and complete Freund's adjuvant (CFA) mixed liquor.Rapamycin (RAPA,2.5 mg · kg-1) was administered by intraperitoneal injection 0.5 h after immunization and capsaicin (1 mg · kg-1 · d-1) was administered by intragastric administration 1.0 h after immunization for 15 days.The incidence and clinical characteristics of EAN were observed.The clinical scores of neurological signs were completed and body weight was measured.Pathological morphology of sciatic nerve was observed by HE staining and Lauck fast blue staining.Ultrastructure of sciatic nerve was observed by transmission electron microscope.Levels of serum tumor necrosis factor alpha (TNF-α),interferon gamma (IFN-γ),interleukin 1β (IL-1β) and intedeukin-6 (IL-6) were tested by enzyme linked immunosorbent assay (ELISA).Expressions of autophagy related protein were measured by Western blot.Results Compared with EAN group,the clinical scores of neurological signs significantly decreased from day 7 to day 16 of post-immunization (P ＜ 0.05),body weight significantly increased from day 3 to day 16 of post-immunization (P ＜ 0.05),demyelination obviously decreased,inflammatory cell infiltration number obviously decreased (P ＜ 0.05),the levels of TNF-α,IFN-γ,IL-1β and IL-6 significantly decreased (P ＜ 0.05),the number of autophagosome in axon of sciatic nerve significantly decreased (P ＜ 0.05),and expressions of Beclin-1 and LC3-Ⅱ and the ratio of LC3-Ⅱ and LC3-Ⅰ were significantly down-regulated,and the expression of p62 was significantly up-regulated (P ＜ 0.05) in EAN + capsaicin group.Rapamycin partially reversed the action of capsaicin.Conclusions Capsaicin inhibits EAN in rats,and the mechanism may be related with the inhibition of autophagy activity.
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Objective To explore expression of Beclin1, autophagy-related protein 1 (Atg1), mammalian target of ra-pamycin (mTOR) in squamous cell carcinoma (SCC) and adenocarcinoma of lung and to analyse relationship among these three factors. Methods Immunohistochemical stainning was applied to detect expression of Beclin1, Atg1 and mTOR in 82 samples of SCC and adenocarcinoma of lung (lung cancer group) and 17 samples of paraneoplastic normal lung tissues (con-trol group). Correlations of expression of Beclin1, Atg1 and mTOR were analyzed in SCC and adenocarcinoma of lung. Re-sults The expression of Beclin1 was significantly lower in lung cancer group than that in control group(52.44%vs 94.12%, P＜0.01). There were significant differences of the differentiation, TNM stages and lymph node metastasis in lung cancer group compared with those in control group. And Beclin1 expression showed a decreasing trend with malignant and invasive of tumors. mTOR expression in lung cancer tissue with lymph nod metastasis is higher than that in the tissue without lymph node metastasis(P＜0.01). Atg1 expression in lung cancer tissue with larger mass is less than that in lung cancer tissue with small lumps(P＜0.01). Beclin1 Expression was positively correlated with the Atg1 expression in SCC(r=0.609,P＜0.01). Beclin1 and Atg1 expressions were both negatively correlated with mTOR expression in SCC(r=-0.617,-0.444,P＜0.01). Beclin1 expression was positively correlated with Atg1 expression in adenocarcinoma(r=0.458,P＜0.01). Beclin1 and Atg1 expression were both negatively correlated with mTOR expression in adenocarcinoma(r=-0.497,-0.541,P＜0.01). Conclu-sion Beclin1, Atg1 and mTOR expression showed a strong correlation in lung cancer, and clinicopathological change of lung SCC and adenocarcinoma may be related to autophagy.
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Objective To investigate the autophagy capacity in clear cell renal cell carcinoma tissue compared with adjacent normal tissue.Methods Sixty-nine pairs of samples from human clear cell renal cell carcinoma tissues and relatively healthy renal tissues adjacent to the tumor were collected during surgical resection.The expressions of proteins that were participating in the regulation and execution of autophagy were detected by immunohistochemisty.Electron-microscopy was also carried out for the morphometrical analysis.Results The protein expression of p-mTOR (P =0.004),P62 (P =0.000) and ULK1 (P =0.000) were up-regulated in the carcinoma tissue,while the expression of Beclin1 (P=0.000),LC3 (P =0.000) and ATG7 (P =0.000) were down-regulated,and the expression of ATG5 (P =0.349) had no signif-icant difference compared with adjacent normal tissue.Morphometrical analysis showed that the basal autophagic activity (measured by the presence of autophagy vacuole compartment) was remarkably down-regulated in carcinoma tissue,compared with adjacent normal tissue.The expression level of mTOR was correlated with P62,LC3 and ATG7,but results showed no correlation between mTOR and Beclin 1.Conclusion Our studies show a relatively reduced autophagy capacity in clear cell renal cell carcinoma,which is regulated by multiple autophagy-related proteins such as mTOR,Beclin 1 and LC3.