العنوان البديل: Hypoxic postconditioning protects myocardium by regulating autophagy in aging cardiomyocytes through piRNA-005854. (English)

المؤلفون: 迟宏扬, 杨慧霞, 郝银菊, 杨安宁, 白志刚, 焦 运, 熊建团, 马胜超, 姜怡邓

المصدر: Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu; 5/8/2024, Vol. 28 Issue 13, p2054-2060, 7p

مصطلحات موضوعية: LACTATE dehydrogenase, ISCHEMIC postconditioning, REPERFUSION injury, MYOCARDIAL injury, WESTERN immunoblotting, GALACTOSE, CREATINE kinase

الملخص (بالإنجليزية): BACKGROUND: Ischemic postconditioning is one of the effective ways to reduce ischemia-reperfusion injury and has been more and more widely used in clinical practice in recent years, but its specific molecular mechanism has yet to be studied. OBJECTIVE: To investigate the role and mechanism of piRNA-005854 in the aging cardiomyocytes caused by hypoxic postconditioning. METHODS: In vitro, cardiomyocytes were administered 8 mg/mL D-galactose for 9 days to induce their aging. β-Galactosidase staining was used to observe the aging of cardiomyocytes. Senescent cells were treated with hypoxia/reoxygenation and hypoxic postconditioning. ELISA was utilized to detect changes in myocardial injury markers creatine kinase isoenzyme MB and lactate dehydrogenase levels. Western blot assay was applied to detect the expression changes of autophagy-related proteins LC3II, p62, ULK1 and phosphorylated ULK1 in aging cardiomyocytes. qRT-PCR was employed to determine the expression level of piRNA-005854. piRNA-005854 inhibitor and piRNA-005854 mimics were transferred into aging cardiomyocytes and followed with hypoxic postconditioning. Western blot assay was used to examine the expression of LC3II, p62, ULK1 and phosphorylated ULK1. RESULTS AND CONCLUSION: (1) D-galactose induced obvious senescence of cardiomyocytes 9 days later. (2) Compared with the normoxia group, creatine kinase isoenzyme MB and lactate dehydrogenase levels increased in the hypoxia/reoxygenation group (P < 0.01); LC3 II/I expression was increased; p62 expression was decreased; ULK1 phosphorylation level was increased, and piRNA-005854 expression was increased (P < 0.01). (3) Compared with the hypoxia/ reoxygenation group, creatine kinase isoenzyme MB and lactate dehydrogenase levels significantly reduced in the hypoxic postconditioning group (P < 0.01); LC3 II/I expression significantly decreased (P < 0.05); p62 expression increased (P < 0.01); ULK1 phosphorylation level decreased (P < 0.05), and piRNA-005854 expression decreased (P < 0.01). (4) After transfection of piRNA-005854 inhibitor, LC3II/I expression was decreased (P < 0.01); the expression of p62 was increased significantly (P < 0.05); the phosphorylation level of ULK1 was decreased significantly (P < 0.01). After transfection of piRNA-005854 mimics, LC3II/ I expression was increased significantly; the expression of p62 was decreased, and the phosphorylation level of ULK1 was increased significantly (P < 0.01). (5) The results show that piRNA-005854-mediated reduction of ULK1-dependent autophagy level is a possible mechanism that hypoxic postconditioning exerts its protective effect on aging cardiomyocytes. [ABSTRACT FROM AUTHOR]

Abstract (Chinese): 背景：缺血后处理是减轻缺血再灌注损伤的有效方式之一, 近年来被越来越广泛地应用于临床实践, 但其具体分子机制还有待研究。 目的：探讨piRNA-005854在衰老心肌细胞缺氧后处理中的作用及机制。 方法：体外给予心肌细胞8 mg/mL D-半乳糖9 d诱导其衰老, β-半乳糖苷酶染色观察心肌细胞的衰老情况；衰老后细胞给予缺氧/复氧处 理和缺氧后处理, ELISA检测心肌损伤标志物肌酸激酶同工酶MB以及乳酸脱氢酶水平；Western blot检测衰老心肌细胞中自噬相关蛋白 LC3Ⅱ、p62和ULK1及其磷酸化ULK1的表达；qRT-PCR检测piRNA-005854的表达水平；进一步用piRNA-005854 inhibitor及piRNA-005854 mimics 转染衰老心肌细胞并进行缺氧后处理, Western blot检测LC3Ⅱ、p62和ULK1及其磷酸化ULK1的表达。 结果与结论：①D-半乳糖诱导9 d后心肌细胞出现明显衰老；②与正常氧组比较, 缺氧/复氧组肌酸激酶同工酶MB以及乳酸脱氢酶水平增 加(P < 0.01)；LC3Ⅱ/Ⅰ表达升高、p62表达降低、ULK1磷酸化水平升高、piRNA-005854表达升高(P < 0.01)；③与缺氧/复氧组比较, 缺氧后 处理组肌酸激酶同工酶MB以及乳酸脱氢酶水平明显减少(P < 0.01)；LC3Ⅱ/Ⅰ表达明显降低(P < 0.05)、p62表达升高(P < 0.01)、ULK1磷酸化 水平降低(P < 0.05)、piRNA-005854表达降低(P < 0.01)；④转染piRNA-005854 inhibitor后, LC3Ⅱ/Ⅰ表达降低(P < 0.01), p62表达明显升高(P < 0.05), ULK1磷酸化水平明显降低(P < 0.01)；转染piRNA-005854 mimics后, LC3Ⅱ/Ⅰ表达显著升高, p62表达降低, ULK1磷酸化水平明显增 加(P < 0.01)；⑤结果表明, piRNA-005854介导的ULK1依赖性自噬水平降低是衰老心肌细胞缺氧后处理发挥保护作用的可能机制。 [ABSTRACT FROM AUTHOR]

: Copyright of Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu is the property of Chinese Journal of Tissue Engineering Research and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

العنوان البديل: Effect of cyclic RNA hsa-circ-0001360 on homocysteine-induced apoptosis of human umbilical vein endothelial cells. (English)

المؤلفون: 况园军, 于素美, 钟颖怡, 章旭红, 马胜超, 杨安宁, 郝银菊, 熊建团, 焦 运, 姜怡邓

المصدر: Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu; 9/8/2024, Vol. 28 Issue 25, p4060-4064, 5p

الملخص (بالإنجليزية): BACKGROUND: Increased homocysteine level induces apoptosis of human umbilical vein endothelial cells, but the mechanism remains unclear. OBJECTIVE: To investigate the role of hsa-circ-0001360 in human umbilical vein endothelial cell apoptosis induced by homocysteine. METHODS: In vitro cultured human umbilical vein endothelial cells were divided into control group, homocysteine group, interference control group, interference control + homocysteine group, hsa-circ-0001360 interference group, hsa-circ-0001360 + homocysteine interference group, overexpression control group, overexpression control + homocysteine group, hsa-circ-0001360 overexpression group and hsa-circ-0001360 + homocysteine overexpression group. All groups were treated with 100 μmol/L homocysteine. After 72 hours of intervention, the expressions of apoptosis-related proteins Bax, Bcl-2, and Caspase-3 were detected by western blot assay. The apoptotic rate was detected by flow cytometry. Quantitative real-time PCR was used to detect the expression of hsacirc-0001360. RESULTS AND CONCLUSION: (1) Compared with the control group, the expression of Caspase-3 and Bax was significantly increased (P < 0.01), and the expression of Bcl-2 was significantly decreased (P < 0.01), and the apoptotic rate was significantly increased (P < 0.01) in the homocysteine group. (2) Compared with control group, the expression of hsa-circ-0001360 was significantly increased in the homocysteine group (P < 0.01). (3) The expression of hsa-circ-0001360 was significantly higher in the cytoplasm than that in the nucleus (P < 0.01). (4) Compared with the interference control C group and interference control + homocysteine group, the expressions of Caspase-3 and Bax were significantly decreased (P < 0.01), while the expression of Bcl-2 was significantly increased (P < 0.01); the apoptotic rate was significantly decreased (P < 0.01) in sh-hsa-circ-0001360 interference group and sh-hsa-circ-0001360 + homocysteine interference group. (5) Compared with overexpression control group and overexpression control + homocysteine group, the expressions of Caspase-3 and Bax were significantly increased (P < 0.01), while the expression of Bcl-2 was significantly decreased (P < 0.01); the apoptotic rate was significantly increased (P < 0.01) in the hsa-circ-0001360 overexpression group and the hsa-circ-0001360 + homocysteine overexpression group. (6) In conclusion, hsa-circ-0001360 can promote the apoptosis of human umbilical vein endothelial cells induced by homocysteine. [ABSTRACT FROM AUTHOR]

Abstract (Chinese): 背景：同型半胱氨酸水平上升会诱导人脐静脉内皮细胞发生凋亡，但机制尚不清楚。 目的：探讨hsa-circ-0001360在同型半胱氨酸诱导人脐静脉内皮细胞发生凋亡中的作用。 方法：体外培养人脐静脉内皮细胞，将其分为对照组、同型半胱氨酸组、干扰对照组、干扰对照+同型半胱氨酸组、干扰hsa-circ-0001360 组、干扰hsa-circ-0001360+同型半胱氨酸组、过表达对照组、过表达对照+同型半胱氨酸组、过表达hsa-circ-0001360组和过表达 hsa-circ-0001360+同型半胱氨酸组，同型半胱氨酸的干预浓度均为100 μmol/L。干预细胞72 h后，应用Western blot检测凋亡相关蛋白Bax、 Bcl-2和Caspase-3的表达；流式细胞仪检测细胞的凋亡率；实时荧光定量PCR(qRT-PCR)检测hsa-circ-0001360的表达水平。 结果与结论：①与对照组相比，同型半胱氨酸组人脐静脉内皮细胞中Caspase-3和Bax的表达明显升高(P < 0.01)，Bcl-2的表达明显降低( P < 0.01)，细胞凋亡率明显增高(P < 0.01)；②与对照组相比，同型半胱氨酸组人脐静脉内皮细胞中hsa-circ-0001360的表达明显升高(P < 0.01)； ③hsa-circ-0001360在人脐静脉内皮细胞的细胞质中表达显著高于细胞核(P < 0.01)；④与干扰对照组或干扰对照+同型半胱氨酸组相比，干 扰hsa-circ-0001360组和干扰hsa-circ-0001360+同型半胱氨酸组人脐静脉内皮细胞中Caspase-3、Bax的表达明显降低(P < 0.01)，Bcl-2的表达明 显升高(P < 0.01)，细胞凋亡率明显降低(P < 0.01)；⑤与过表达对照组或过表达对照+同型半胱氨酸组相比，过表达hsa-circ-0001360组和过 表达hsa-circ-0001360+同型半胱氨酸组人脐静脉内皮细胞中Caspase-3、Bax的表达明显升高(P < 0.01)，Bcl-2的表达明显降低(P < 0.01)，细胞 凋亡率明显升高(P < 0.01)；⑥以上结果表明，hsa-circ-0001360可以促进同型半胱氨酸诱导人脐静脉内皮细胞发生凋亡。 [ABSTRACT FROM AUTHOR]

: Copyright of Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu is the property of Chinese Journal of Tissue Engineering Research and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

العنوان البديل: Involvement of miR-144-3p in Cbs^+/- mouse hepatocyte autophagy induced by high-methionine diet. (English)

المؤلفون: 盛思琪, 谢 琳, 赵翔宇, 姜怡邓, 吴 凯, 熊建团, 杨安宁, 郝银菊, 焦 运

المصدر: Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu; 3/18/2024, Vol. 28 Issue 8, p1289-1294, 6p

مصطلحات موضوعية: FATTY liver, AUTOIMMUNE hepatitis, PEARSON correlation (Statistics), CELL survival, GENE knockout, ASPARTATE aminotransferase

الملخص (بالإنجليزية): BACKGROUND: High-methionine diet can cause liver injury in Cbs^+/- mice, and hyperhomocystinemia is related to the occurrence and progression of various liver-related diseases, such as hepatic steatosis, autoimmune hepatitis, and alcoholic fatty liver disease. MicroRNAs (miRNAs) are involved in various cellular processes including cell survival, differentiation and autophagy, which are of great significance. OBJECTIVE: To investigate the critical role of miR-144-3p on Cbs^+/- mouse hepatocyte autophagy induced by high methionine die. METHODS: (1) Ten male cystathione-β-synthase normal (Cbs^+/+) mice and another 10 male mice with single gene knockout (Cbs^+/-) of similar body mass, 4 weeks of age, were fed a high-methionine diet and executed after 12 weeks to take liver tissue. (2) Human hepatocytes (HL-7702) were cultured in vitro and divided into control [0 μmol/L homocysteine (Hcy)], Hcy (100 μmol/L Hcy), mimic-NC (transfected with mimic-NC), mimic-NC + Hcy (mimic-NC transfecton+100 μmol/L Hcy), miR-144-3p mimic (transfected with miR-144-3p mimic), and miR-144-3p mimic + Hcy (miR-144-3p mimic transfection+100 μ mol/L Hcy), inhibitor-NC (transfected with inhibitor-NC), inhibitor-NC + Hcy (inhibitor-NC transfection + 100 μmol/L Hcy), miR-144-3p inhibitor (transfected with miR-144-3p inhibitor), and miR-144-3p inhibitor + Hcy (miR-144-3p inhibitor transfection + 100 μmol/L Hcy). Quantitative real-time PCR was used to detect the expression of miR144-3p in liver tissue and hepatocytes. After transfection of miR-144-3p mimic or inhibitor, quantitative real-time PCR and western blot were used to detect the transfection efficiency of miR-144-3p and its effect on the expression of autophagy-related proteins LC3B and p62. The levels of alanine transferase and aspartate aminotransferase in hepatocyte supernatants were determined by enzyme linked immunosorbent assay. The correlation between the expression of miR-144-3 in hepatocyte and the levels of alanine transferase and aspartate aminotransferase in hepatocyte supernatants were analyzed by Pearson correlation analysis. RESULTS AND CONCLUSION: Compared with the Cbs^+/+ group and control group, the expression of miR-144-3p in the liver tissue of the Cbs^+/- group and in hepatocytes of the Hcy group was decreased (P < 0.01). The expression of LC3B-II/I was decreased in hepatocyte after transfection of miR-144-3p mimic, while the protein expression of p62 was increased (P < 0.01). The opposite results were obtained after transfection of miR-144-3p inhibitor (P < 0.01). Compared with the mimic-NC group, the levels of alanine transferase and aspartate aminotransferase were decreased in the miR-144-3p mimic group (P < 0.01), while the opposite results were obtained in the inhibitor-NC group (P < 0.01). The expression of miR-144-3p in hepatocytes was negatively correlated with the levels of alanine transferase (P < 0.01, r=-0.887 6) and aspartate aminotransferase (P < 0.01, r=-0.829 9) in the supernatant of hepatocytes. To conclude, Hcy promotes hepatocyte autophagy by inhibiting the expression of miR-144-3p, which subsequently aggravates liver injury. [ABSTRACT FROM AUTHOR]

Abstract (Chinese): 背景：高蛋氨酸饮食可导致Cbs^+/- 小鼠发生肝损伤, 高同型半胱氨酸血症与肝脂肪变性、自身免疫性肝炎、酒精性脂肪肝等多种肝脏相关 疾病的发生和进展有关。微小RNA (miRNAs)参与细胞存活、分化和细胞自噬等各种细胞过程, 具有重要意义。 目的：探讨miR-144-3p在高蛋氨酸饮食诱导Cbs^+/- 小鼠肝细胞自噬中的关键作用。 方法：①选取4周龄体质量相近的雄性胱硫醚β-合成酶基因正常(Cbs^+/+)小鼠和单基因敲除(Cbs^+/- )小鼠各10只, 均饲以高蛋氨酸饮食, 12 周后处死, 留取肝脏组织。②体外培养人源肝细胞(HL-7702), 分为对照组(0 μmol/L同型半胱氨酸)、同型半胱氨酸组(100 μmol/L同型半 胱氨酸)、mimic-NC组(转染mimic-NC)、mimic-NC+同型半胱氨酸组(转染mimic-NC+100 μmol/L同型半胱氨酸)、miR-144-3p mimic组(转染 miR-144-3p mimic)、miR-144-3p mimic+同型半胱氨酸组(转染miR-144-3p mimic+100 μmol/L同型半胱氨酸)、inhibitor-NC组(转染inhibitor-NC)、 inhibitor-NC+同型半胱氨酸组(转染inhibitor-NC+100 μmol/L同型半胱氨酸)、miR-144-3p inhibitor组(转染miR-144-3p inhibitor)、miR-144-3p inhibitor+同型半胱氨酸组(转染miR-144-3p inhibitor+100 μmol/L同型半胱氨酸)。采用荧光定量PCR检测肝组织和肝细胞中miR-144-3p的表达 水平；转染miR-144-3p模拟物或抑制剂后, 采用荧光定量PCR和Western blot分别检测miR-144-3p的转染效率及其对LC3B和p62蛋白表达的影 响；酶联免疫法检测肝细胞上清液中丙氨酸氨基转移酶和门冬氨酸氨基转移酶的表达情况；Pearson相关性分析肝细胞miR-144-3p表达与肝 细胞上清液中丙氨酸氨基转移酶和门冬氨酸氨基转移酶含量的相关性。 结果与结论：①与Cbs^+/+组比较, Cbs^+/- 组小鼠肝组织和同型半胱氨酸组肝细胞中miR-144-3p的表达水平降低(P < 0.01)；②转染miR-144-3p 模拟物后, 与mimic-NC比较, miR-144-3p mimic组中LC3B-Ⅱ/Ⅰ蛋白的表达水平降低, p62蛋白的表达水平升高(P < 0.01)；转染miR-144-3p inhibitor后, 得到相反的结果(P < 0.01)；③与mimic-NC组相比, miR-144-3p mimic组肝细胞上清液中丙氨酸氨基转移酶和门冬氨酸氨基转移 酶的含量降低(P < 0.01)；与inhibitor-NC组比较, 得到相反的结果(P < 0.01)；④肝细胞中miR-144-3p的表达与肝细胞上清液中丙氨酸氨基转 移酶(P < 0.01, r=-0.887 6)和门冬氨酸氨基转移酶(P < 0.01, r=-0.829 9)的含量呈负相关；⑤结果表明, 同型半胱氨酸通过抑制miR-144-3p 的表达促进肝细胞的自噬, 进而加重肝损伤。 [ABSTRACT FROM AUTHOR]

: Copyright of Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu is the property of Chinese Journal of Tissue Engineering Research and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

العنوان البديل: Shikonin inhibits fibrosis of human hypertrophic scar fibroblasts via MicroRNA-382-5p. (English)

المؤلفون: 唐玉婷, 贺 茜, 万 瑀, 王建军, 杨安宁, 吴 凯, 焦 运, 白志刚, 姜怡邓, 沈江涌

المصدر: Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu; 12/18/2023, Vol. 27 Issue 35, p5642-5648, 7p

مصطلحات موضوعية: HYPERTROPHIC scars, SCARS, DIMETHYL sulfone, GENE expression, CELL migration, CELL culture, COLLAGEN

الملخص (بالإنجليزية): BACKGROUND: Previous studies have shown that shikonin has the potential to treat hypertrophic scar and that microRNAs are involved in the regulation of the pathological mechanism of hypertrophic scar. It is speculated that shikonin may regulate the occurrence and development of hypertrophic scar through microRNAs regulation. OBJECTIVE: To investigate the mechanism of shikonin on the fibrosis of human hypertrophic scar fibroblasts via MicroRNA-382-5p. METHODS: Hypertrophic scar tissue and normal skin tissue adjacent to the scar (within 3 cm around the scar) were provided by the Department of Burn and Plastic Surgery, General Hospital of Ningxia Medical University to extract human hypertrophic scar fibroblasts and human normal skin fibroblasts, respectively. Hematoxylin-eosin staining was used to identify normal skin and hypertrophic scar and immunofluorescence was used to identify fibroblasts. The relative expression level of MicroRNA-382-5p was detected by quantitative real-time PCR at the tissue level. Hypertrophic scar fibroblasts were randomly divided into shikonin group (shikonin was dissolved in dimethyl sulfone to make drug solution at a concentration of 13.46 μmol/L, which was used for cell culture for 24 hours), dimethyl sulfone group, MicroRNA-382-5p negative control group, MicroRNA-382-5p inhibitor group, shikonin+MicroRNA-382-5p negative control group and shikonin+MicroRNA-382-5p overexpression group. Real-time fluorescence quantitative PCR and western blot were used to detect the expression of Collagen Type I α1, Collagen Type III α1 and α-smooth muscle actin at mRNA and protein levels, respectively. Cell counting kit-8 was used to detect cell viability. Cell scratch test was used to detect the migration ability of cells. RESULTS AND CONCLUSION: Compared with normal skin fibroblasts, hypertrophic scar fibroblasts had stronger proliferative activity (P < 0.01). Compared with the dimethyl sulfone group, shikonin down-regulated the expression levels of Collagen Type I α1, Collagen Type III α1 and α-smooth muscle actin in human hypertrophic scar fibroblasts (mRNA: P < 0.01, protein: P < 0.01), and inhibited the migration of hypertrophic scar fibroblasts (P < 0.05). Compared with normal skin, MicroRNA-382-5p was highly expressed in hypertrophic scar (P < 0.01), and shikonin could down-regulate the expression of MicroRNA-382-5p (P < 0.01). Inhibition of MicroRNA-382-5p down-regulated the expression level of Collagen Type I α1, Collagen Type III α1 and α-smooth muscle actin in hypertrophic scar fibroblasts (mRNA: P < 0.01, protein: P < 0.01), and inhibited the migration of hypertrophic scar fibroblasts (P < 0.05). Under the influence of shikonin, overexpression of MicroRNA-382-5p upregulated the expression levels of Collagen Type I α1, Collagen Type III α1 and α-smooth muscle actin in hypertrophic scar fibroblasts (mRNA: P < 0.01, protein: P < 0.01), and promoted the migration of hypertrophic scar fibroblasts (P < 0.01). To conclude, shikonin can inhibit the fibrosis and migration of hypertrophic scar fibroblasts by down-regulating the expression of MicroRNA-382-5p. [ABSTRACT FROM AUTHOR]

Abstract (Chinese): 背景：目前已有研究表明紫草素具有治疗增生性瘢痕的潜力,且miRNA参与增生性瘢痕的病理机制调控,猜测紫草素有可能通过影响 miRNA调控增生性瘢痕的发生发展. 目的：探讨紫草素通过影响MicroRNA-382-5p对人增生性瘢痕成纤维细胞纤维化的作用机制. 方法：收集宁夏医科大学总医院烧伤整形外科提供的增生性瘢痕组织及瘢痕旁正常皮肤组织(瘢痕旁3 cm以内),并分别分离出人增生性瘢 痕成纤维细胞和正常成纤维细胞用于后续实验.苏木精-伊红染色鉴定正常皮肤和增生性瘢痕；免疫荧光鉴定成纤维细胞；采用实时荧光 定量PCR从组织水平检测MicroRNA-382-5p相对表达水平.将增生性瘢痕成纤维细胞随机分为紫草素组(紫草素溶于二甲基亚砜配成浓度为 13.46 μmol/L的药物干预细胞24 h),二甲基亚砜组,MicroRNA-382-5p阴性对照组,MicroRNA-382-5p抑制组,紫草素+MicroRNA-382-5p阴性 对照组及紫草素+MicroRNA-382-5p过表达组.实时荧光定量PCR检测Ⅰ型胶原蛋白α1,Ⅲ型胶原蛋白α1和α-平滑肌肌动蛋白mRNA表达； Western blot检测Ⅰ型胶原蛋白α1,Ⅲ型胶原蛋白α1和α-平滑肌肌动蛋白的蛋白表达；CCK-8法检测细胞活性；划痕实验检测细胞迁移能力. 结果与结论：①与正常皮肤成纤维细胞相比,增生性瘢痕成纤维细胞增殖活性更强(P < 0.01)；②与二甲基亚砜组相比,紫草素组可以下调增 生性瘢痕成纤维细胞中Ⅰ型胶原蛋白α1,Ⅲ型胶原蛋白α1和α-平滑肌肌动蛋白的表达水平(mRNA：P < 0.01,蛋白：P < 0.01),并抑制增生性 瘢痕成纤维细胞迁移(P < 0.05)；③与正常皮肤相比,MicroRNA-382-5p在增生性瘢痕中呈高表达(P < 0.01),且紫草素能下调增生性瘢痕成纤维 细胞中MicroRNA-382-5p的表达(P < 0.01)；④敲低MicroRNA-382-5p能下调增生性瘢痕成纤维细胞中Ⅰ型胶原蛋白α1,Ⅲ型胶原蛋白α1和α-平滑 肌肌动蛋白的表达水平(mRNA：P < 0.01,蛋白：P < 0.01),并抑制增生性瘢痕成纤维细胞迁移(P < 0.05)；⑤过表达MicroRNA-382-5p能上调在紫 草素影响下增生性瘢痕成纤维细胞中Ⅰ型胶原蛋白α1,Ⅲ型胶原蛋白α1和α-平滑肌肌动蛋白的表达水平(mRNA：P < 0.01,蛋白：P < 0.01),并促 进增生性瘢痕成纤维细胞迁移(P < 0.01)；⑥提示紫草素可通过下调MicroRNA-382-5p的表达抑制增生性瘢痕成纤维细胞的纤维化和迁移. [ABSTRACT FROM AUTHOR]

: Copyright of Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu is the property of Chinese Journal of Tissue Engineering Research and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

العنوان البديل: Erastin inhibits proliferation of hypertrophic scar fibroblasts. (English)

المؤلفون: 贺 茜, 万 瑀, 唐玉婷, 杨安宁, 吴 凯, 焦 运, 白志刚, 姜怡邓, 沈江涌

المصدر: Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu; 11/18/2023, Vol. 27 Issue 32, p5120-5125, 6p

مصطلحات موضوعية: PROLIFERATING cell nuclear antigen, HYPERTROPHIC scars, CYCLIN-dependent kinase inhibitors, FERRITIN, GENE expression, IRON ions

الملخص (بالإنجليزية): BACKGROUND: Hypertrophic scar is a kind of pathological scar that appears in the healing process after skin trauma caused by various reasons and there is no effective treatment. OBJECTIVE: To investigate the effect of ferroptosis inducer (Erastin) on the proliferation of human hypertrophic scar fibroblasts. METHODS: Hypertrophic scar samples provided by the Burn Plastic Surgery Department of the General Hospital of Ningxia Medical University and normal skin samples of the same individual were collected. Human hypertrophic scar fibroblasts were then extracted for subsequent experiments. (1) The cells were divided into control group (without treatment) and ferroptosis inducer group (treated with 20 μmol/L Erastin for 24 hours). The expression of Ferritin was detected by western blot. Iron ion detection kit was used to measure cellular iron ion concentration. Malondialdehyde detection kit was used to detect cellular malondialdehyde content. (2) The cells were divided into control group (without treatment), ferroptosis inducer group (treated with 20 μmol/L Erastin for 24 hours) and ferroptosis inducer+ferroptosis inhibitor group (co-treated with 20 μmol/L Erastin and 20 μmol/L Ferrostatin-1 for 24 hours). The mRNA and protein expressions of proliferating cell nuclear antigen (PCNA) and cyclin-dependent kinase inhibitor (p27) were detected using qRT-PCR and western blot. Cell counting kit-8 and EdU were used to detect cell proliferation viability and levels. RESULTS AND CONCLUSION: Compared with the control group, Erastin decreased the ferritin expression (P < 0.01), increased the content of iron ions (P < 0.05), and elevated the malondialdehyde content in the cells (P < 0.01). Compared with the control group, Erastin decreased the expression of PCNA (P < 0.01), increased the expression of p27 (P < 0.05), weakened the cell proliferation ability (P < 0.01), and reduced the number of EdU-positive cells (P < 0.01). Compared with the ferroptosis inducer group, the ferroptosis inducer + ferroptosis inhibitor group had increased expression of PCNA (P < 0.05), decreased p27 expression (P < 0.05), enhanced cell proliferation (P < 0.01), and increased number of EdU-positive cells (P < 0.05). To conclude, the ferroptosis inducer (Erastin) induces ferroptosis in hypertrophic scar fibroblasts and then inhibits their proliferation. [ABSTRACT FROM AUTHOR]

Abstract (Chinese): 背景：增生性瘢痕是各种原因导致的皮肤创伤后愈合过程中出现的一种病理性瘢痕，目前缺乏特效治疗方法。 目的：探讨铁死亡诱导剂Erastin对人增生性瘢痕成纤维细胞增殖的影响。 方法：收集宁夏医科大学总医院烧伤整形外科提供的增生性瘢痕组织和同一个体正常皮肤，提取人增生性瘢痕成纤维细胞进行后续实验。 将细胞分为对照组(不做处理)和铁死亡诱导剂组(用20 μmol/L的Erastin干预细胞24 h)，Western blot检测铁蛋白(Ferritin)的表达，铁离子检 测试剂盒测定细胞铁离子浓度，丙二醛检测试剂盒检测细胞丙二醛水平；将细胞分为对照组(不做处理)、铁死亡诱导剂组(用20 μmol/L的 Erastin干预细胞24 h)和铁死亡诱导剂+铁死亡抑制剂组(用20 μmol/L的Erastin和20 μmol/L的Ferrostatin-1同时干预细胞24 h)，qRT-PCR检测 PCNA及p27的mRNA表达，Western blot检测PCNA及p27的蛋白表达，CCK-8、EdU检测细胞增殖活力和增殖水平。 结果与结论：①与对照组相比，铁死亡诱导剂组铁蛋白减少(P < 0.01)，铁离子增多(P < 0.05)，丙二醛增多(P < 0.01)；②与对照组相比，铁 死亡诱导剂组PCNA的表达降低(P < 0.01)，p27的表达增加(P < 0.05)，细胞增殖能力减弱(P < 0.01)，EdU阳性细胞数减少(P < 0.01)；③与铁死 亡诱导剂组相比，铁死亡诱导剂+铁死亡抑制剂组PCNA的表达增加(P < 0.05)，p27的表达降低(P < 0.05)，细胞增殖能力增强(P < 0.01)，EdU 阳性细胞数增多(P < 0.05)；④结果表明，铁死亡诱导剂Erastin通过诱导增生性瘢痕成纤维细胞发生铁死亡进而抑制其增殖。 [ABSTRACT FROM AUTHOR]

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العنوان البديل: Role of LncRNA MALAT1 in myocardial autophagy reduction in aging rats after ischemic postconditioning. (English)

المؤلفون: 杨慧霞, 揭育祯, 白志刚, 焦 运, 杨 勇, 马天龙, 马胜超, 姜怡邓

المصدر: Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu; 7/18/2023, Vol. 27 Issue 20, p3173-3179, 7p

مصطلحات موضوعية: LINCRNA, ISCHEMIC postconditioning, SPRAGUE Dawley rats, REPERFUSION injury, PROTEIN expression

الملخص (بالإنجليزية): BACKGROUND: Ischemic postconditioning can alleviate myocardial ischemia-reperfusion injury, but the specific mechanism is not clear. OBJECTIVE: To investigate the mechanism of long non-coding RNA (lncRNA) MALAT1 in the reduction of autophagy levels in aging myocardium induced by ischemic postconditioning. METHODS: Twenty-seven Sprague-Dawley rats aged 22-24 months were randomly divided into three groups, with nine rats in each group: sham operation, ischemia-reperfusion, and ischemic postconditioning groups. Morphological changes of myocardial tissue were observed by hematoxylin-eosin staining and Masson staining. Rat myocardial cells (H9C2) were induced in vitro with 8 mg/mL D-galactose for 9 days and then divided into normoxia, hypoxia-reoxygenation, and hypoxia postconditioning groups. Western blot was used to detect the protein expression levels of LC3II/I and p62. Fluorescence quantitative PCR was used to detect the expression of lncRNA MALAT1 in aging myocardium and aging cardiomyocytes. Autophagy double-labeled adenovirus (RFP-GFP-LC3) was used to observe the changes of autophagic flux in aging cardiomyocytes. lncRNA MALAT1 interference fragment and overexpression plasmid were transfected into aging cardiomyocytes and the protein expression levels of LC3II/I and p62 were detected by western blot. RESULTS AND CONCLUSION: Compared with the ischemia-reperfusion group, the myocardial tissue structure of the ischemic postconditioning group was basically clear, the nucleus was intact, and the deposition of blue collagen fibers in the myocardial tissue was reduced. Compared with the ischemia-reperfusion group, the expression of LC3II/I was decreased and the expression of p62 was increased in the ischemic postconditioning group (P < 0.05). Compared with the hypoxia-reperfusion group, the expression of LC3II/I was decreased (P < 0.01) and the expression of p62 was increased (P < 0.05) in the hypoxia postconditioning group, and the number of intracellular autophagosomes and autophagolysosomes was decreased (P < 0.05). Compared with the ischemiareperfusion group, the expression of MALAT1 in the aging myocardial tissue was decreased the ischemic postconditioning group (P < 0.01); compared with the hypoxia-reperfusion group, the expression of MALAT1 in aging cardiomyocytes was decreased in the hypoxic postconditioning group (P < 0.01). Compared with the hypoxia postconditioning+si-NC group, the expression of LC3II/I was decreased and the expression of p62 was increased in the hypoxia postconditioning +si-lncRNA MALAT1 (P < 0.01); compared with the hypoxia postconditioning+ad-NC group, the expression of LC3II/I was increased and the expression of p62 was decreased in the hypoxia postconditioning+ad-lncRNA MALAT1 group (P < 0.01). To conclude, the lncRNA MALAT1 mediated reduction of autophagy levels is an important mechanism underlying the protective effect of ischemic postconditioning in aging myocardium. [ABSTRACT FROM AUTHOR]

Abstract (Chinese): 背景：缺血后处理可缓解心肌缺血再灌注损伤，但是其具体机制尚不清楚。 目的：探讨lncRNA MALAT1在缺血后处理所引起衰老心肌自噬水平降低中的作用。 方法： 27只22-24月龄SD大鼠随机分为3组：假手术组、缺血再灌注组和缺血后处理组，每组9只，采用苏木精-伊红染色和Masson染色 观察心肌组织形态学变化； 体外使用8 mg/mL D-半乳糖诱导大鼠心肌(H9C2)细胞9 d后，分为正常氧组、缺氧复氧组和缺氧后处理组。 Western blot检测衰老心肌组织及衰老心肌细胞中LC3Ⅱ/Ⅰ和p62蛋白的表达；采用qRT-PCR检测衰老心肌组织和衰老心肌细胞中MALAT1相 对表达；转染自噬双标腺病毒(RFP-GFP-LC3)观察衰老心肌细胞自噬流的变化；衰老心肌细胞转染MALAT1干扰片段和过表达质粒，Western blot检测各组细胞中LC3Ⅱ/Ⅰ和p62蛋白的表达。 结果与结论： 与缺血再灌注组比较，缺血后处理组心肌组织结构基本清晰，细胞核完整，心肌组织间蓝色胶原纤维沉积减少； 与缺 血再灌注组比较，缺血后处理组LC3Ⅱ/Ⅰ表达降低且p62表达增高(P < 0.05)； 与缺氧复氧组比较，缺氧后处理组LC3Ⅱ/Ⅰ表达降低(P < 0.01)且p62表达增加(P < 0.05)，细胞内自噬体和自噬溶酶体数量均减少(P < 0.01)； 与缺血再灌注组比较，缺血后处理组衰老心肌组织的 MALAT1表达降低(P < 0.01)；与缺氧复氧组比较，缺氧后处理组衰老心肌细胞的MALAT1表达降低(P < 0.01)； 衰老心肌细胞转染MALAT1 干扰片段和过表达质粒后，与缺氧复氧+si-NC组比较，缺氧复氧+si-MALAT1组LC3Ⅱ/Ⅰ表达降低且p62表达增加(P < 0.01)；与缺氧后处理+ ad-NC组比较，缺氧后处理+ad-MALAT1组LC3Ⅱ/Ⅰ表达增加且p62表达降低(P < 0.01)； 结果表明：lncRNA MALAT1介导的自噬水平降低是 衰老大鼠心肌缺血后处理发挥保护作用的重要机制。 [ABSTRACT FROM AUTHOR]

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العنوان البديل: High efficient flame retardancy and smoke suppression effect of cobalt hydroxystannate on flexible polyvinyl chloride. (English)

المؤلفون: 胡伟东, 赵贺, 焦运红, 吴静

المصدر: Acta Materiae Compositae Sinica; Sep2019, Vol. 36 Issue 9, p2067-2075, 9p

مصطلحات موضوعية: HEAT release rates, POLYVINYL chloride, FIREPROOFING agents, TENSILE tests, FLAME, COMBUSTION, FLAMMABILITY

الملخص (بالإنجليزية): Submicrometer-sized cobalt hydroxystannate (CHS) flame retardant was prepared via coprecipitation method and applied to flexible polyvinyl chloride (PVC) to obtained CHS/PVC composites. The flame retardancy, thermal stability and mechanical properties of CHS/PVC composites were investigated by limiting oxygen index (LOI), cone calorimeter, TG analysis and universal tensile tests. The results show that the CHS effectively enhances the flame retardancy and maintains the mechanical property of CHS/PVC composites. Compared with blank PVC, when the mass ratio of CHS to PVC is 10%, the LOI of CHS/PVC composite is increased by 2.3%, the peaks of heat release rate and smoke production rate are reduced by 39.6% and 57.4%, respectively. The enhancement for CHS/PVC composites in flame retardation mainly attributes to the water dehydration from CHS when heating which dilutes heat. On the other hand, the CoCl2 generated during the combustion process can effectively catalyze the early decomposition of PVC and then a denser and continuous residual charlayer is formed, thereby the combustion of PVC is effectively suppress. [ABSTRACT FROM AUTHOR]

Abstract (Chinese): 通过共沉淀法制备了亚微米尺寸的羟基锡酸钴（CHS）阻燃剂，并将其应用于软质聚氯乙烯（PVC）中，制得CHS/PVC复合材料。采用极限氧指数仪（LOI）、锥形量热仪、TG和拉伸仪研究了CHS/PVC复合材料的阻燃性能、热稳定性和力学性能。结果表明，CHS可以有效提高CHS/PVC复合材料的阻燃性能，并对CHS/PVC复合材料的力学性能保护较好；与空白PVC相比，当CHS添加量（CHS与PVC质量比）为10%时，CHS/PVC复合材料的LOI增加了2.3%，热释放速率峰值和烟释放速率峰值分别下降了39.6%和57.4%。这主要是由于CHS受热后脱除的水具有冷却和稀释热量的作用；另一方面在燃烧过程中生成的CoCl2可以有效催化PVC早期分解，形成更加致密且连续的残炭，从而有效抑制PVC的燃烧。 [ABSTRACT FROM AUTHOR]

: Copyright of Acta Materiae Compositae Sinica is the property of Acta Materiea Compositae Sinica Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

العنوان البديل: Therapeutic Effects of Anti-CD90 Monoclonal Antibody in Melanoma.

المؤلفون: 焦运燊¹ jiaoyunshen@163.com, 沈光前¹, 初明¹ famous@bjmu.edu.cn, 张苗苗¹, 王颖¹, 丁羚昱¹, 李欢欢¹, 蔡如意¹, 王月丹¹ wangyuedan@bjmu.edu.cn

المصدر: Progress in Modern Biomedicine. 10/10/2017, Vol. 17 Issue 28, p5401-5405. 5p.

الملخص (بالإنجليزية): Objective: To investigate the feasibility and mechanism of anti-CD90 monoclonal antibody in the treatment of melanoma. Methods: hi this study, the possibility of anti-CD90 monoclonal antibody to induce the B16 cell line apoptosis in vitro was analyzed by flow cytometry firstly. Afterwards, the murine melanoma model was established by subcutaneously injecting B16 cells into C57BL/6J mice, then evaluated the anti-tumor effect of anti-CD90 monoclonal antibody by comparing the size and weight of the tumors. The formation and distribution of neovascularization in tumor tissues were detected by iinmunohistochemistry, for comparing the microvascular density in the control group and the therapy group. Results: Although anti-CD90 monoclonal antibody could not directly induce the apoptosis of B16 cells, it could inhibit the progress of melanoma in mice in situ (p=0.049), and the neovascularization in therapy group was significantly reduced. The t-test results showed that there was a significant difference in microvascular density between the treatment group and the control group (p=0.011). Conclusions: Anti-C'D90 monoclonal antibody can inhibit the development of melanoma by inhibiting the formation of neovascularization in tumor tissue, so as to acliieve the effect of melanoma treatment. [ABSTRACT FROM AUTHOR]

Abstract (Chinese): 目的:探讨应用抗CD90 单克隆抗体治疗黑色素细胞瘤的可行性及相关机制。方法：首先通过流式细胞检测技术，探究抗 CD90 单克隆抗体是否能在体外诱导B16 细胞凋亡。之后，通过向C57BL/6J 小鼠皮下注射B16 细胞建立小鼠黑色素瘤模型，并 给予抗CD90 单克隆抗体治疗，评价其抗肿瘤的治疗效果。同时，应用免疫组织化学的方法观察小鼠成瘤组织中新生血管的形成 和分布情况，统计肿瘤中微血管密度进行比较。结果：抗CD90单克隆抗体在体外不能直接诱导B16 细胞凋亡，但抗CD90 单克隆 抗体能够抑制小鼠黑色素瘤的原位生长（p=0.049）；使用免疫组织化学染色法对肿瘤组织切片进行染色，发现经过抗体治疗的小 鼠成瘤组织中的新生血管明显减少，治疗组和对照组肿瘤组织的微血管密度存在显著性差异（p = 0.011）。结论：抗CD90单克隆 抗体能够通过抑制肿瘤组织中新生血管的形成影响黑色素瘤的发生发展，从而达到治疗黑色素瘤的效果。 [ABSTRACT FROM AUTHOR]

العنوان البديل: The Expression of TFDP3 in Breast Cancer and Its Role of Breast Cancer Molecular Classification. (English)

المؤلفون: 陈雪, 初明, 杨珂, 阴凯麟, 何嘉勰, 徐兰, 焦运燊, 丁羚昱, 刘彦辰, 初正云, 王月丹

المصدر: Progress in Modern Biomedicine; 2017, Vol. 17 Issue 1, p1-6, 6p

الملخص (بالإنجليزية): Objective: The object of this study is to analysis the expression of cancer testis antigen TFDP3 in breast cancer, to explore the correlation between TFDP3 and development of breast cancer. Methods: The expression of TFDP3 was detected in pathological section of tumor tissues from patients with breast cancer using immunohistochemical method. And we analyzed the data with clinical information of patients to discover the correlation of TFDP3 and the molecular classification of breast cancer. We also detected the expression of TFDP3 protein in five breast cancer cell lines, including MDA-MB-231, MCF-7, SK-BR-3,T47D and MCF-10A, using Western Blot.Then we verified the location of TFDP3 protein in cells through immumofluorescence method. Results: TFDP3 is greatly and broadly expressed in many cell lines and breast cancer tissues, such as breast, lymph node,hypothyroid, appendix and so on, while TFDP3 is 49％ percent positive in breast cancer. The expression of TFDP3 is unrelated with clinical stage, pathology type and molecular classification of breast cancer, but related with special tumor marker HER2. Conclusion: TFDP3 highly expresses in breast cancer is related with the expression of HER2. It suggests that TFDP3 be a new potential target molecule of immunotherapy for breast cancer [ABSTRACT FROM AUTHOR]

Abstract (Chinese): 摘要： 目的:本研究旨在分析总结癌-睾丸抗原TFDP3在乳腺癌中的表达规律,探究TFDP3表达与乳腺癌分型及发病进程关系.方法:通过对不同分型及分期的乳腺癌组织切片进行免疫组化检测,分析TFDP3在其中的表达情况,并结合样本来源患者的临床信息,对TFDP3的表达与乳腺癌分子分型、分期及预后的相关性进行统计分析.同时,通过Western Blot检测了5种乳腺癌细胞系(MDA-MB-231、MCF-7、SK-BR-3、T47D及MCF-10A)中TFDP3的表达情况,并通过细胞免疫荧光实验,检测TFDP3在细胞中的定位.结果:TFDP3在已检测的乳腺癌样本中的表达率为49％.乳腺癌临床分子分型标记物HER2的表达与TFDP3的表达呈正相关,但乳腺癌的分期、临床病理分型以及临床分子分型标记物ER、PR的表达均与肿瘤组织中TFDP3的表达无相关性.结论:TFDP3在各在乳腺癌组织中高表达,其表达量与HER2的表达量呈正性相关.这为研究已TFDP3为靶点的乳腺癌免疫治疗,提供了新的依据. [ABSTRACT FROM AUTHOR]

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