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Role and mechanism of phosphate myosin light chain in chronic allograft nephropathy of rats
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作者 王玉新 《外科研究与新技术》 2011年第4期281-281,共1页
Objective To investigate role and mechanism of phosphate myosin light chain ( pMLC) in rat kidney of chronic allograft nephropathy ( CAN) model. Methods Left donor kidneys from Fisher ( F344) rats were ortho-topically... Objective To investigate role and mechanism of phosphate myosin light chain ( pMLC) in rat kidney of chronic allograft nephropathy ( CAN) model. Methods Left donor kidneys from Fisher ( F344) rats were ortho-topically transplanted into Lewis recipients,Meanwhile, F344 rats and LEW rats with resection of right 展开更多
关键词 Role and mechanism of phosphate myosin light chain in chronic allograft nephropathy of rats ILK
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Preliminary research on myosin light chain kinase in rabbit liver 被引量:6
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作者 Bin Ren~1 Hua-Qing Zhu~2 Zhao-Feng Luo~1 Qing Zhou~2 Yuan Wang~2 Yu-Zhen Wang~1 1 Department of Biochemistry and Molecular Biology,University of Science and Technology of China,Hefei 230027,Anhui Province,China2 Laboratory of Molecular Biology,Anhui Medical University,Hefei 230032,Anhui Province,China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第6期868-871,共4页
AIM: To study preliminarily the properties of myosin light chain kinase (MLCK) in rabbit liver. METHODS: The expression of MLCK was detected by reverse transcription-polymerase chain reaction(RT-PCR); the MLCK was obt... AIM: To study preliminarily the properties of myosin light chain kinase (MLCK) in rabbit liver. METHODS: The expression of MLCK was detected by reverse transcription-polymerase chain reaction(RT-PCR); the MLCK was obtained from rabbit liver, and its activity was analyzed by gamma-(32)P incorporation technique to detect the phosphorylation of myosin light chain. RESULTS: MLCK was expressed in rabbit liver, and the activity of the enzyme was similar to rabbit smooth muscle MLCK, and calmodulin-dependent. When the concentration was 0.65 mg x L(-1), the activity was at the highest level. CONCLUSION: MLCK expressed in rabbit liver may catalyze the phosphorylation of myosin light chain, which may play important roles in the regulation of hepatic cell functions. 展开更多
关键词 ANIMALS HEPATOCYTES Liver myosin light chains myosin-light-chain Kinase PHOSPHORYLATION RABBITS Research Support Non-U.S. Gov't
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Epidermal Growth Factor Enhances Orthovanadate-Induced Contraction via Src and Myosin Phosphatase Target Subunit 1 in Rat Vascular Smooth Muscle
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作者 Tomoya Sasahara Natsumi Ohkura +2 位作者 Mariko Shin Akira Onodera Katsutoshi Yayama 《Pharmacology & Pharmacy》 2015年第7期329-340,共12页
Inhibition of protein tyrosine phosphatase by orthovanadate induces vasoconstriction, which is mediated by the Rho kinase-dependent inactivation of myosin light chain phosphatase (MLCP) via signaling downstream of Src... Inhibition of protein tyrosine phosphatase by orthovanadate induces vasoconstriction, which is mediated by the Rho kinase-dependent inactivation of myosin light chain phosphatase (MLCP) via signaling downstream of Src-induced activation of the epidermal growth factor (EGF) receptor. The present study investigated the potential role of EGF in orthovanadate (OVA)-dependent vaso-constriction. OVA-induced aortic contraction significantly increased in the presence of EGF, and was abolished by inhibitors of Rho kinase (Y27632), extracellular signal-regulated kinase 1 and 2 (Erk1/2) (FR180204), Erk1/2 kinase (PD98059), EGF receptor (AG1478), and Src (PP2). Treatment of the rat endothelium-denuded thoracic aorta with either EGF or OVA augmented the phosphorylation of myosin phosphatase target subunit 1 (MYPT1) at Thr-853 and of the EGF receptor at Tyr-1173. The phosphorylation of MYPT1 was further increased by co-stimulation with EGF and OVA. EGF receptor phosphorylation at Tyr-845 was also increased by EGF or OVA;this effect was augmented by co-stimulation with EGF and OVA, and was abolished by Src inhibition. In addition, Erk1/2 was phosphorylated by EGF or by co-treatment with EGF and OVA;this was abolished by an EGF receptor inhibitor, but not by Src inhibition. These results suggested that OVA-induced EGF-related contraction was mediated by the Rho kinase-dependent inactivation of MLCP via two different signaling cascades: Src-dependent phosphorylation of the EGF receptor at Tyr-845 and EGF-dependent phosphorylation of Erk1/2. 展开更多
关键词 EPIDERMAL Growth Factor myosin light chain PHOSPHATASE MITOGEN-ACTIVATED Kinase ORTHOVANADATE
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Adaptation of Skeletal Muscle to Prolonged Activity: Role of Myosin
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作者 Teet Seene Karin Alev Priit Kaasik 《Health》 2019年第2期195-200,共6页
The aim of this short review is to describe the role of myosin isoforms during the adaptation of skeletal muscle to prolonged physical activity (for example endurance exercise) and to show the coordination between cha... The aim of this short review is to describe the role of myosin isoforms during the adaptation of skeletal muscle to prolonged physical activity (for example endurance exercise) and to show the coordination between changes in muscle oxidative capacity and myofibrillar apparatus in slow-twitch and fast-twitch muscles. Adaptational changes in myosin isoforms during long lasting muscle activity (decrease of MyHC IIb isoforms relative content and increase of that MyHC IIa and decrease of MyLC 1 fast isoforms in fast-twitch muscles) are in good coordination with changes of muscle oxidative capacity. These changes show that during regular endurance exercise fast-twitch muscle fibers (type IIA) are also recruited and create the potential source of increase in endurance capacity during the process of adaptation to the prolonged physical activity. 展开更多
关键词 PROLONGED Muscle ACTIVITY ENDURANCE Exercise myosin light chain myosin Heavy chain ENDURANCE Capacity
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柴胡皂苷A调节MLCK/MLC2信号通路对SAP大鼠肠损伤的影响
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作者 孙兵 陶韬 《国际检验医学杂志》 CAS 2024年第4期462-466,共5页
目的探讨柴胡皂苷A调节肌球蛋白轻链激酶(MLCK)/肌球蛋白轻链2(MLC2)信号通路对重症急性胰腺炎(SAP)大鼠肠损伤的影响。方法随机选择10只大鼠作为假手术组,其余大鼠注射牛磺胆酸钠溶液构建SAP大鼠模型。将造模成功的SAP大鼠模型随机平分... 目的探讨柴胡皂苷A调节肌球蛋白轻链激酶(MLCK)/肌球蛋白轻链2(MLC2)信号通路对重症急性胰腺炎(SAP)大鼠肠损伤的影响。方法随机选择10只大鼠作为假手术组,其余大鼠注射牛磺胆酸钠溶液构建SAP大鼠模型。将造模成功的SAP大鼠模型随机平分为SAP组、柴胡皂苷A组(腹腔注射10.0 mg/kg的柴胡皂苷A)、iE-DAP组(腹腔注射3.5 mg/kg MLCK/MLC2通路激活剂iE-DAP)、柴胡皂苷A+iE-DAP组(腹腔注射10.0 mg/kg柴胡皂苷A+3.5 mg/kg iE-DAP),每组均10只大鼠,每天1次,连续注射1周,假手术组和SAP组注射等量生理盐水。酶联免疫吸附试验(ELISA)检测各组大鼠血清淀粉酶(AMY)、脂肪酶(LIP)、二胺氧化酶(DAO)、白细胞介素(IL)-1β、IL-6和肿瘤坏死因子-α(TNF-α)水平;HE染色检测各组大鼠回肠组织病理形态变化。ELISA检测各组大鼠回肠组织中氧化应激指标水平。蛋白质免疫印迹检测肠道屏障相关蛋白及MLCK/MLC2通路相关蛋白表达。结果与SAP组相比,柴胡皂苷A组AMY、LIP、DAO、IL-1β、IL-6和TNF-α水平显著降低,而iE-DAP组AMY、LIP、DAO、IL-1β、IL-6和TNF-α水平显著升高,差异有统计学意义(P<0.05)。与SAP组相比,柴胡皂苷A组大鼠回肠组织结构得到改善,回肠组织病理评分显著降低(P<0.05)。与SAP组相比,柴胡皂苷A组谷胱甘肽、超氧化物歧化酶水平显著升高,丙二醛水平显著降低,差异有统计学意义(P<0.05)。与SAP组相比,柴胡皂苷A组MLCK、p-MLC2/MLC2蛋白水平显著降低,差异有统计学意义(P<0.05)。结论柴胡皂苷A可能通过下调MLCK/MLC2信号通路对SAP大鼠肠损伤起到改善作用。 展开更多
关键词 柴胡皂苷A 肌球蛋白轻链激酶/肌球蛋白轻链2信号通路 重症急性胰腺炎 肠损伤
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生血通便颗粒对血虚肠燥型慢传输型便秘大鼠结肠肌电及Ca^(2+)/CaM/MLCK信号通路的影响 被引量:1
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作者 罗雯鹏 王真权 +2 位作者 周佳敏 肖俐敏 王军文 《中国中医药信息杂志》 CAS CSCD 2024年第2期97-103,共7页
目的观察生血通便颗粒对血虚肠燥型慢传输型便秘(STC)大鼠结肠肌电及Ca^(2+)/CaM/MLCK信号通路的影响,探讨其治疗STC的作用机制。方法采用洛哌丁胺灌胃结合尾部放血法建立血虚肠燥型STC大鼠模型。将大鼠分为对照组、模型组、枸橼酸莫沙... 目的观察生血通便颗粒对血虚肠燥型慢传输型便秘(STC)大鼠结肠肌电及Ca^(2+)/CaM/MLCK信号通路的影响,探讨其治疗STC的作用机制。方法采用洛哌丁胺灌胃结合尾部放血法建立血虚肠燥型STC大鼠模型。将大鼠分为对照组、模型组、枸橼酸莫沙必利组和生血通便颗粒组,每组8只,给药组分别予相应药物灌胃,连续14 d。观察大鼠治疗前后一般状况,检测大鼠粪便含水量,生物机能实验系统检测结肠肌电慢波频率、振幅及变异系数,测定肠道推进率,HE染色观察结肠组织病理变化,比色法检测结肠平滑肌细胞Ca^(2+)浓度,Western blot检测结肠平滑肌组织缝隙连接蛋白43(Cx43)、钙调蛋白(CaM)、肌球蛋白轻链激酶(MLCK)、磷酸化肌球蛋白轻链20(p-MLC20)蛋白表达。结果与对照组比较,模型组大鼠体质量、粪便含水量和肠道推进率显著降低(P<0.01),结肠肌电慢波频率减慢、频率变异系数增加(P<0.01),慢波振幅和振幅变异系数增加(P<0.01);结肠黏膜结构受损,可见炎性改变,糜烂明显,结肠平滑肌细胞内Ca^(2+)浓度及Cx43、CaM、MLCK、p-MLC20蛋白表达显著降低(P<0.01)。与模型组比较,枸橼酸莫沙必利组和生血通便颗粒组大鼠体质量、粪便含水量和肠道推进率显著升高(P<0.05,P<0.01),结肠肌电慢波频率加快、频率变异系数减小(P<0.01),慢波振幅和振幅变异系数减小(P<0.05,P<0.01);结肠黏膜结构较完整,糜烂情况改善,结肠平滑肌细胞内Ca^(2+)浓度及Cx43、CaM、MLCK、p-MLC20蛋白表达显著升高(P<0.01)。结论生血通便颗粒可改善血虚肠燥型STC大鼠症状,恢复结肠动力,其机制可能与调节结肠肌电节律性、激活Ca^(2+)/CaM/MLCK信号通路相关。 展开更多
关键词 生血通便颗粒 慢传输型便秘 钙离子 钙调蛋白 肌球蛋白轻链激酶
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补中益气方通过MLCK/p-MLC_(20)通路调节慢性腹泻大鼠结肠动力的机制研究
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作者 贾梦迪 张声生 +2 位作者 赵鲁卿 卢小芳 朱泠霏 《辽宁中医杂志》 CAS 北大核心 2024年第10期180-186,共7页
目的探讨补中益气方对慢性腹泻大鼠MLCK/p-MLC_(20)通路介导的结肠动力的影响。方法大鼠随机分为正常组,模型组和补中益气低、中、高剂量组。模型组及补中益气方组应用大黄灌胃的方法建立慢性腹泻大鼠模型。慢性腹泻模型建立成功后,补... 目的探讨补中益气方对慢性腹泻大鼠MLCK/p-MLC_(20)通路介导的结肠动力的影响。方法大鼠随机分为正常组,模型组和补中益气低、中、高剂量组。模型组及补中益气方组应用大黄灌胃的方法建立慢性腹泻大鼠模型。慢性腹泻模型建立成功后,补中益气汤颗粒低剂量:0.4725 g/(kg·d),中剂量:0.945 g/(kg·d),高剂量:1.89 g/(kg·d),正常组和模型组则每天给予等体积的生理盐水灌胃。在治疗结束时,检测各组大鼠一般情况如体质量、饮食量、悬空拉尾抵抗实验、免疫指标(胸腺指数、脾脏指数)变化和24小时粪便含水量。进一步探讨机制,采用离体张力测定灌流系统,测定各组大鼠结肠纵行平滑肌肌条收缩力(colonic longitudinal smooth muscle strips,CLSMs)和结肠环型平滑肌肌条(colonic circular smooth muscle strips,CCSMs)的变化,并蛋白印迹法测定磷酸化肌球蛋白轻链(phosphorylation level of myosin light chain,p-MLC_(20))水平及实时荧光定量PCR法检测大鼠结肠组织肌球蛋白轻链激酶(myosin light chain kinase,MLCK)的表达。结果各组大鼠一般情况比较有统计学意义(P<0.05),与正常组比较,模型组一般情况表现为(体质量下降:P<0.01;饮食量减少:P<0.01;悬空拉尾抵抗实验时间缩短:P<0.01;胸腺指数降低:P<0.01;脾脏指数降低:P<0.01),与模型组比较,补中益气低、中、高剂量组一般情况明显改善(体质量增加:低剂量:P<0.05,中、高剂量:P<0.01;饮食量增多:低剂量:P<0.05,中、高剂量:P<0.01;悬空拉尾抵抗实验时间延长:低剂量:P<0.05,中、高剂量:P<0.01;胸腺指数升高:P<0.01;脾脏指数升高:P<0.01)。各组大鼠粪便含水量差异有统计学意义(P<0.01),与正常组比较,模型组粪便含水量明显增加(P<0.01),与模型组比较,补中益气中、高剂量组粪便含水量明显下降(P<0.01)。同时,正常组、模型组和补中益气中剂量组大鼠结肠CLSMs和CCSMs收缩差异有统计学意义(P<0.05),与正常组比较,模型组大鼠CLSMs和CCSMs收缩张力、振幅、速率明显增加(P<0.01),与模型组比较,补中益气中剂量组大鼠CLSMs和CCSMs收缩张力、振幅、速率有所下降(P<0.05);三组大鼠结肠p-MLC_(20)含量差异有统计学意义(P<0.05),与正常组比较,模型组大鼠p-MLC_(20)含量明显增加(P<0.01),与模型组比较,补中益气中剂量组大鼠p-MLC_(20)含量明显下降(P<0.01);三组大鼠结肠MLCK的表达差异有统计学意义(P<0.01),与正常组比较,模型组大鼠MLCK的表达升高(P<0.01),与模型组比较,补中益气中剂量组MLCK的表达降低(P<0.01)。结论补中益气方对慢性腹泻大鼠治疗作用可能是通过调节MLCK/p-MLC20通路抑制结肠动力来实现的。 展开更多
关键词 慢性腹泻 补中益气方 动力 肌球蛋白轻链 肌球蛋白轻链激酶
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肌球蛋白轻链激酶在恶性肿瘤中的研究进展
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作者 杨哲宇 肖瑞 +1 位作者 王瑞 任建军 《中国医药》 2024年第11期1756-1760,共5页
近年来,肌球蛋白轻链激酶(MLCK)作为细胞骨架和细胞运动的重要调节因子,在多种肿瘤的发生发展中发挥着关键作用。MLCK通过细胞信号转导,参与调控肿瘤细胞的增殖、迁移、侵袭和凋亡等生物过程。鉴于MLCK在肿瘤发展中的关键作用,其作为肿... 近年来,肌球蛋白轻链激酶(MLCK)作为细胞骨架和细胞运动的重要调节因子,在多种肿瘤的发生发展中发挥着关键作用。MLCK通过细胞信号转导,参与调控肿瘤细胞的增殖、迁移、侵袭和凋亡等生物过程。鉴于MLCK在肿瘤发展中的关键作用,其作为肿瘤治疗的潜在靶点被广泛研究,旨在开发针对MLCK的治疗策略,以提高治疗效率和患者生存质量。本文综述了MLCK在不同类型肿瘤中的表达模式、功能机制及其潜在的临床应用价值,同时探讨了MLCK作为潜在治疗靶点的研究现状及未来发展方向,为临床治疗提供新的思路和策略。 展开更多
关键词 肿瘤 肌球蛋白轻链激酶 细胞迁移 细胞骨架 治疗靶点
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circRNA MYLK基因干扰对胰腺癌PANC-1细胞线粒体膜电位、氧化损伤和微管形成的影响
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作者 李青聪 黄鑫 +2 位作者 李自康 李燕 罗威 《联勤军事医学》 CAS 2024年第1期11-16,共6页
目的探讨circRNA肌球蛋白轻链激酶(myosin light chain kinase,MYLK)基因干扰对胰腺癌PANC-1细胞线粒体膜电位、氧化损伤和微管形成的影响。方法将对数生长期PANC-1细胞分为空白对照组、shRNA-NC组和circMYLK-shRNA组。转染后,采用细胞... 目的探讨circRNA肌球蛋白轻链激酶(myosin light chain kinase,MYLK)基因干扰对胰腺癌PANC-1细胞线粒体膜电位、氧化损伤和微管形成的影响。方法将对数生长期PANC-1细胞分为空白对照组、shRNA-NC组和circMYLK-shRNA组。转染后,采用细胞克隆形成实验检测各组PANC-1细胞的生长。试剂盒检测各组细胞上清液中超氧化物歧化酶(superoxide dismutase,SOD)和丙二醛(malondialdehyde,MDA)水平。流式细胞仪分析细胞线粒体膜电位。显微镜下观察各组细胞微管结节数。Western blot检测各组细胞Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)/B细胞淋巴瘤2基因(B-cell lymphoma 2,Bcl-2)、原癌基因c-Myc、血管内皮生长因子(vascular endothelial growth factor,VEGF)和波形蛋白(Vimentin)表达。结果与shRNA-NC组相比,circMYLK-shRNA1组PANC-1细胞的克隆形成率、JC-1红色荧光所占百分比、微管结节数明显降低(P均<0.05);细胞上清中SOD水平明显降低,MDA水平明显升高(P均<0.05);细胞内Bax/Bcl-2蛋白表达明显升高(P<0.05),c-Myc、VEGF和Vimentin蛋白表达明显降低(P均<0.05)。结论circRNA MYLK基因沉默可以抑制PANC-1细胞增殖能力,降低PANC-1细胞的线粒体膜电位,诱导细胞氧化损伤,抑制微管的形成。 展开更多
关键词 circRNA肌球蛋白轻链激酶 胰腺癌 线粒体膜电位 氧化损伤 微管
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非肌型肌球蛋白轻链激酶的结构、功能及其生理与病理学意义
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作者 林亦婷 林燕琴 +2 位作者 韦锦填 陈健 黄丽 《中国组织化学与细胞化学杂志》 CAS CSCD 2024年第1期76-80,共5页
肌球蛋白轻链激酶(myosin light-chain kinase,MYLK)在哺乳动物中由MYLK基因编码,编码的产物主要包括长链非肌型肌球蛋白轻链激酶(non-muscle myosin light-chain kinase,nm MYLK,220 kDa),短链平滑肌型肌球蛋白轻链激酶(smooth muscle ... 肌球蛋白轻链激酶(myosin light-chain kinase,MYLK)在哺乳动物中由MYLK基因编码,编码的产物主要包括长链非肌型肌球蛋白轻链激酶(non-muscle myosin light-chain kinase,nm MYLK,220 kDa),短链平滑肌型肌球蛋白轻链激酶(smooth muscle myosin light-chain kinase,sm MYLK,130 kDa)及端蛋白(telokin)(17 kDa)。其中nm MYLK的特殊N端结构域赋予其参与更多病理生理学过程的特性,包括炎症、细胞黏附、肿瘤细胞浸润、动脉粥样斑块形成等。本文综述nmMYLK的结构、功能研究进展,梳理nmMYLK与炎症、癌症及其他疾病间的关系。 展开更多
关键词 非肌型肌球蛋白轻链激酶 肿瘤 炎症
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MYLIP和miR-542-3p与宫颈癌患者早期诊断及临床预后的关系
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作者 田文 尹晓梅 +4 位作者 付淼 刘蓬 王莎 刘昊 王东海 《中国妇幼健康研究》 2024年第10期55-62,共8页
目的探讨血清肌球蛋白调节轻链相互作用蛋白(MYLIP)和微小RNA-542-3p(miR-542-3p)表达水平对宫颈癌(CC)早期诊断价值及与其临床预后的关系。方法选取本院2018年5月至2020年5月期间收治的128例CC患者作为CC组,另选取同期于本院就诊的86... 目的探讨血清肌球蛋白调节轻链相互作用蛋白(MYLIP)和微小RNA-542-3p(miR-542-3p)表达水平对宫颈癌(CC)早期诊断价值及与其临床预后的关系。方法选取本院2018年5月至2020年5月期间收治的128例CC患者作为CC组,另选取同期于本院就诊的86例宫颈上皮瘤变(CIN)患者作为CIN组,行体检的74例体检健康者作为对照组。采用实时荧光定量逆转录聚合酶链反应(qRT-PCR)法检测各组血清MYLIP mRNA和miR-542-3p表达水平,收集CC患者临床病理特征资料,分析二者表达水平与CC患者临床病理特征的关系;使用ENCORI网站预测MYLIP和miR-542-3p之间是否存在靶向关系;使用Pearson法分析MYLIP mRNA和miR-542-3p表达的相关性;绘制受试者工作特征(ROC)曲线评估MYLIP mRNA和miR-542-3p表达水平对CC的早期诊断价值;使用Kaplan-Meier法分析血清MYLIP mRNA和miR-542-3p表达水平与CC患者预后的关系;使用Cox回归分析影响CC患者预后的风险因素。结果CC组血清MYLIP mRNA表达水平显著高于CIN组和对照组(F=118.850,P<0.001),CC组血清miR-542-3p表达水平显著低于CIN组和对照组(F=85.413,P<0.001);CC患者血清MYLIP mRNA和miR-542-3p表达水平与是否绝经(χ^(2)=5.026、6.133)、宫颈浸润深度(χ^(2)=6.767、7.957)、FIGO分期(χ^(2)=7.788、11.480)、TNM分期(χ^(2)=12.426、20.150)、淋巴结是否转移(χ^(2)=8.821、12.687)有关(P<0.05);MYLIP和miR-542-3p存在靶向结合位点,且CC患者血清MYLIP mRNA与miR-542-3p表达为负相关关系(r=-0.415,P<0.001);血清MYLIP mRNA和miR-542-3p表达水平单独及联合诊断CC的曲线下面积(AUC)分别为0.807、0.815、0.894,联合诊断效果优于单独诊断(Z二者联合-MYLIP=2.165、P=0.030;Z二者联合-miR-542-3p=2.360,P=0.018);血清MYLIP mRNA高表达患者3年生存率显著低于低表达患者(52.24%vs.78.69%,χ^(2)=10.869,P=0.001),血清miR-542-3p高表达患者3年生存率显著高于低表达患者(79.03%vs.51.52%,χ^(2)=11.771,P=0.001);MYLIP mRNA高表达(HR=1.972,95%CI:1.565~2.485)是CC患者死亡的危险因素,miR-542-3p高表达(HR=0.517,95%CI:0.399~0.670)是CC患者死亡的保护因素(P<0.05)。结论血清MYLIP mRNA和miR-542-3p表达水平对早期诊断宫颈癌具有一定的价值,且二者表达水平与宫颈癌患者临床预后密切相关。 展开更多
关键词 肌球蛋白调节轻链相互作用蛋白 微小RNA-542-3p 宫颈癌 早期诊断 临床预后
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地衣芽孢杆菌制剂对非酒精性脂肪性肝病大鼠模型肝脂肪变性及肠黏膜屏障功能的影响
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作者 赵春燕 李逸群 李莉 《临床肝胆病杂志》 CAS 北大核心 2024年第10期2000-2007,共8页
目的通过建立非酒精性脂肪性肝病(NAFLD)大鼠模型,探讨地衣芽孢杆菌制剂对大鼠肝组织病理学及肠黏膜屏障功能的影响。方法将30只雄性SD大鼠随机分为正常对照组(Control组,n=5)、模型组(Mod组,n=15)、地衣芽孢杆菌低剂量组(BLL组,n=5)和... 目的通过建立非酒精性脂肪性肝病(NAFLD)大鼠模型,探讨地衣芽孢杆菌制剂对大鼠肝组织病理学及肠黏膜屏障功能的影响。方法将30只雄性SD大鼠随机分为正常对照组(Control组,n=5)、模型组(Mod组,n=15)、地衣芽孢杆菌低剂量组(BLL组,n=5)和地衣芽孢杆菌高剂量组(BLH组,n=5)。Control组给予普通饲料喂养,Mod组、BLL组、BLH组给予高脂饲料喂养16周,其中BLL组、BLH组在高脂饲料喂养8周后,分别予地衣芽孢杆菌制剂灌胃2.5×10^(7)CFU/kg、5.0×10^(7)CFU/kg,每天1次,持续8周。8周干预结束后,检测大鼠血清ALT、AST、TC、TG、空腹血糖(FPG)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙二醛(MDA)、IL-6、IL-1β和TNF-α水平。HE染色观察肝组织病理学改变并根据NAFLD活动度积分(NAS)评分。透射电镜下观察肠黏膜紧密连接结构。免疫组化染色观察肠黏膜肌球蛋白轻链激酶(MLCK)的表达。实时定量PCR检测肠黏膜MLCK的mRNA水平。高通量16S rRNA测序法分析肠道菌群组成。采用单因素方差分析进行多组间比较,多重比较采用Bonferroni法。结果与Mod组相比,BLH组和BLL组大鼠血清ALT、AST、TC、TG、FPG、IL-1β及TNF-α水平降低,SOD水平升高,肝细胞脂肪变减轻,NAS评分降低,肠黏膜紧密连接结构恢复,MLCK蛋白及mRNA表达水平降低,差异均有统计学意义(P值均<0.05)。此外,BLH组较Mod组CAT水平升高,MDA及IL-6水平下降,差异均有统计学意义(P值均<0.05)。与BLL组相比,BLH组大鼠血清MDA、IL-6、TNF-α水平降低,CAT水平升高,肠黏膜MLCK蛋白及mRNA表达水平降低,差异均有统计学意义(P值均<0.05)。肠道菌群示BLH组和BLL组厚壁菌门和拟杆菌门的相对丰度均较Mod组有所恢复,且BLH组厚壁菌门和拟杆菌门的相对丰度更接近Control组。结论地衣芽孢杆菌制剂可有效减轻NAFLD大鼠的肝脂肪变性,其作用机制可能与下调肠黏膜MLCK蛋白的表达水平、改善肠黏膜紧密连接结构有关。 展开更多
关键词 非酒精性脂肪性肝病 地衣芽孢杆菌 肠黏膜 肌球蛋白轻链激酶
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LncRNA MYLK-AS1调节miR-141-3p/STMN1轴对胃癌细胞增殖、凋亡和侵袭的影响
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作者 刘洁 谢兴明 +1 位作者 钮洪霞 杨先智 《重庆医科大学学报》 CAS CSCD 北大核心 2024年第3期276-282,共7页
目的:探讨长链非编码RNA(long non-coding RNA,LncRNA)肌球蛋白轻链激酶反义RNA1(myosin light chain kinase antisense RNA1,MYLK-AS1)调节miR-141-3p/微管不稳定蛋白1(stathmin 1,STMN1)轴对胃癌细胞增殖、凋亡和侵袭的影响。方法:将H... 目的:探讨长链非编码RNA(long non-coding RNA,LncRNA)肌球蛋白轻链激酶反义RNA1(myosin light chain kinase antisense RNA1,MYLK-AS1)调节miR-141-3p/微管不稳定蛋白1(stathmin 1,STMN1)轴对胃癌细胞增殖、凋亡和侵袭的影响。方法:将HGC27细胞分为NC组、si-NC组、si-MYLK-AS1组、si-MYLK-AS1+inhibitor NC组、si-MYLK-AS1+miR-141-3p inhibitor组。双萤光素酶报告基因实验检测LncRNA MYLK-AS1、miR-141-3p、STMN1的关系;qRT-PCR检测HGC27细胞中LncRNA MYLK-AS1、miR-141-3p表达;CCK-8法检测HGC27细胞增殖情况;流式细胞术检测HGC27细胞凋亡;使用Transwell实验评估了HGC27细胞的侵袭和迁移能力,并统计了穿透基质膜的细胞数量;同时采用Western blot技术检测了HGC27细胞中STMN1、E-cadherin、Vimentin及N-cadherin这几种蛋白表达量的变化。结果:HGC27细胞中LncRNA MYLK-AS1、STMN1水平高于GES-1细胞(P<0.05),miR-141-3p水平低于GES-1细胞(P<0.05)。si-MYLK-AS1组HGC27细胞A_(450 nm)值、迁移、侵袭细胞数量、LncRNA MYLK-AS1表达量、STMN1、N-cadherin、Vimentin蛋白水平低于NC组、si-NC组(P<0.05),HGC27细胞凋亡率、miR-141-3p表达量、E-cadherin蛋白水平高于NC组、si-NC组(P<0.05);而miR-141-3p低表达减弱了沉默LncRNA MYLK-AS1抑制HGC27细胞发展的作用;LncRNA MYLK-AS1靶向调节miR-141-3p/STMN1轴。结论:LncRNA MYLK-AS1可能通过上调microRNA-141-3p的表达水平,间接导致STMN1基因表达受到抑制,这一过程可能对胃癌细胞的增殖、凋亡及侵袭特性产生明显影响。 展开更多
关键词 长链非编码RNA肌球蛋白轻链激酶反义RNA1 微小RNA-141-3p/微管不稳定蛋白1轴 胃癌 增殖 凋亡 侵袭
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CHANGES OF MYOSIN LIGHT CHAIN PATTERNS IN MUSCLE FIBERS AFTER CROSS-REINNERVATION 被引量:2
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作者 任惠民 章生艮 《Chinese Science Bulletin》 SCIE EI CAS 1990年第18期1570-1572,共3页
In the course of analysis of single fibers randomly excided from the fast and slow muscles of the rat for myosin light chain patterns, we have found that the light chains of all fibers taken from extensor digitorum lo... In the course of analysis of single fibers randomly excided from the fast and slow muscles of the rat for myosin light chain patterns, we have found that the light chains of all fibers taken from extensor digitorum longus (EDL) show the same as but different from those of fibers taken from soleus (Sol) in pattern. This characteristic differentiation of 展开更多
关键词 cross-reinnervation myosin light chain.
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Recombinant angiopoietin-like protein 4 attenuates intestinal barrier structure and function injury after ischemia/reperfusion 被引量:4
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作者 Zi-Yi Wang Jian-Yu Lin +8 位作者 Yang-Rong Feng De-Shun Liu Xu-Zi Zhao Tong Li Si-Yuan Li Jing-Chao Sun Shu-Feng Li Wen-Yan Jia Hui-Rong Jing 《World Journal of Gastroenterology》 SCIE CAS 2021年第32期5404-5423,共20页
BACKGROUND Intestinal barrier breakdown,a frequent complication of intestinal ischemiareperfusion(I/R)including dysfunction and the structure changes of the intestine,is characterized by a loss of tight junction and e... BACKGROUND Intestinal barrier breakdown,a frequent complication of intestinal ischemiareperfusion(I/R)including dysfunction and the structure changes of the intestine,is characterized by a loss of tight junction and enhanced permeability of the intestinal barrier and increased mortality.To develop effective and novel therapeutics is important for the improvement of outcome of patients with intestinal barrier deterioration.Recombinant human angiopoietin-like protein 4(rhANGPTL4)is reported to protect the blood-brain barrier when administered exogenously,and endogenous ANGPTL4 deficiency deteriorates radiationinduced intestinal injury.AIM To identify whether rhANGPTL4 may protect intestinal barrier breakdown induced by I/R.METHODS Intestinal I/R injury was elicited through clamping the superior mesenteric artery for 60 min followed by 240 min reperfusion.Intestinal epithelial(Caco-2)cells and human umbilical vein endothelial cells were challenged by hypoxia/reoxygenation to mimic I/R in vitro.RESULTS Indicators including fluorescein isothiocyanate-conjugated dextran(4 kilodaltons;FD-4)clearance,ratio of phosphorylated myosin light chain/total myosin light chain,myosin light chain kinase and loss of zonula occludens-1,claudin-2 and VE-cadherin were significantly increased after intestinal I/R or cell hypoxia/reoxygenation.rhANGPTL4 treatment significantly reversed these indicators,which were associated with inhibiting the inflammatory and oxidative cascade,excessive activation of cellular autophagy and apoptosis and improvement of survival rate.Similar results were observed in vitro when cells were challenged by hypoxia/reoxygenation,whereas rhANGPTL4 reversed the indicators close to normal level in Caco-2 cells and human umbilical vein endothelial cells significantly.CONCLUSION rhANGPTL4 can function as a protective agent against intestinal injury induced by intestinal I/R and improve survival via maintenance of intestinal barrier structure and functions. 展开更多
关键词 Angiopoietin-like protein 4 Intestinal ischemia/reperfusion COVID-19 myosin light chain kinase Intestinal barrier breakdown
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5DFRXXL region of long myosin light chain kinase causes F-actin bundle formation 被引量:2
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作者 YANG Chunxiang WEI Dongmei +2 位作者 CHEN Chen YU Weiping ZHU Minsheng 《Chinese Science Bulletin》 SCIE EI CAS 2005年第18期2044-2050,共7页
Long myosin light chain kinase (L-MLCK) contains five DFRXXL motifs with ability to bind F-actin. Binding stoichiometry data indicated that each DFRXXL motif might bind each G-actin, but its biological significance re... Long myosin light chain kinase (L-MLCK) contains five DFRXXL motifs with ability to bind F-actin. Binding stoichiometry data indicated that each DFRXXL motif might bind each G-actin, but its biological significance remained unknown. We hypothesized that L-MLCK might act as an F-actin bundle peptides by its multiple binding sites of 5DFRXXL motifs to actin. In order to characterize F-actin-bundle formation properties of 5DFRXXL region of long myosin light chain kinase, we expressed and purified 5DFRXXL peptides tagged with HA in vitro. The properties of 5DFRXXL peptides binding to myofilaments or F-actin were analyzed by binding stoichiometries assays. The results indicated that 5DFRXXL peptides bound to myofilaments or F-actin with high affinity. KD values of 5DFRXXL binding to myofilaments and F-actin were 0.45 and 0.41 μmol/L, re- spectively. Cross-linking assay demonstrated that 5DFRXXL peptides could bundle F-actin efficiently. Typical F-actin bundles were observed morphologically through determina- tion of confocal and electron microscopy after adding 5DFRXXL peptides. After transfection of pEGFP-5DFRXXL plasmid into eukaryocyte, spike structure was observed around cell membrane edge. We guess that such structure formation may be attributable to F-actin over-bundle forma- tion caused by 5DFRXXL peptides. Therefore, we suppose that L-MLCK may be a new bundling protein and somehow play a certain role in organization of cell skeleton besides mediating cell contraction by it kinase activity. 展开更多
关键词 肌浆球蛋白链 致活酶 肌丝 DFRXXL 生物学
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The sphingosine-1-phosphate/RhoA/Rho associated kinases/myosin light chain pathway in detrusor of female rats is down-regulated in response to ovariectomy 被引量:1
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作者 Wei Zhang Xiao-Dong Liu +3 位作者 Jia-Wen Wang Ling-Feng Meng Yao-Guang Zhang Jian-Ye Wang 《Chinese Medical Journal》 SCIE CAS CSCD 2020年第10期1203-1210,共8页
Background:Dysuria is one of the main symptoms of genitourinary syndrome of menopause,which causes serious disruption to the normal life of peri-menopausal women.Studies have shown that it is related to decrease of de... Background:Dysuria is one of the main symptoms of genitourinary syndrome of menopause,which causes serious disruption to the normal life of peri-menopausal women.Studies have shown that it is related to decrease of detrusor contractile function,but the exact mechanism is still poorly understood.Previous results have suggested that the sphingosine-1-phosphate(S1P)pathway can regulate detrusor contraction,and this pathway is affected by estrogen in various tissues.However,how estrogen affects this pathway in the detrusor has not been investigated.In this study,we detected changes of the S1P/RhoA/Rho associated kinases(ROCK)/myosin light chain(MLC)pathway in the detrusor of ovariectomized rats in order to explore the underlying mechanism of dysuria during peri-menopause.Methods::Thirty-six female Sprague-Dawley rats were randomly divided into SHAM(sham operation),OVX(ovariectomy),and E groups(ovariectomy+estrogen),with 12 rats in each group.We obtained bladder detrusor tissues from each group and examined the mRNA and protein levels of the major components of the S1P/RhoA/ROCK/MLC pathway using quantitative real-time polymerase chain reaction and Western blotting,respectively.We also quantified the content of S1P in the detrusor using an enzyme linked immunosorbent assay.Finally,we compared results between the groups with one-way analysis of variance.Results::The components of the S1P pathway and the RhoA/ROCK/MLC pathway of the OVX group were significantly decreased,as compared with SHAM group.The percent decreases of the components in the S1P pathway were as follows:sphingosine kinase 1(mRNA:39%,protein:45%)(both P<0.05),S1P(21.73±1.09 nmol/g vs.18.86±0.69 nmol/g)(P<0.05),and S1P receptor 2/3(S1PR2/3)(mRNA:25%,27%,respectively)(P<0.05).However,the protein expression levels of S1PR2/3 and the protein and mRNA levels of SphK2 and S1PR1 did not show significant differences between groups(P>0.05).The percent decreases of the components in the RhoA/ROCK/MLC pathway were as follows:ROCK2(protein:41%,mRNA:36%)(both P<0.05),p-MYPT1(protein:54%)(P<0.05),and p-MLC20(protein:47%)(P<0.05),but there were no significant differences in the mRNA and protein levels of RhoA,ROCK1,MYPT1,and MLC20(all P>0.05).In addition,all of the above-mentioned decreases could be reversed after estrogen supplementation(E group vs.SHAM group)(all P>0.05).Conclusion::In this study,we confirmed that ovariectomy is closely associated with the down-regulation of the S1P/RhoA/ROCK/MLC pathway in the rat detrusor,which may be one mechanism of dysuria caused by decreased contractile function of the female detrusor during peri-menopause. 展开更多
关键词 myosin light chains OVARIECTOMY Rats Rho-associated kinases Sphingosine-1-phosphate Urinary bladder
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子宫肌瘤组织中MMP-2、MLCK、SULT1A3表达及与子宫肌瘤发生的相关性 被引量:5
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作者 王建华 龚钿 张继旺 《中国计划生育学杂志》 2023年第2期429-433,共5页
目的:分析基质金属蛋白酶-2(MMP-2)、肌球蛋白轻链激酶(MLCK)和硫酸基转移酶1A3(SULT1A3)在子宫肌瘤组织中的表达及与子宫肌瘤发生发展的可能关系。方法:收集2019年1月-2021年6月在本院行手术切除治疗的子宫肌瘤患者60例子宫肌瘤组织和... 目的:分析基质金属蛋白酶-2(MMP-2)、肌球蛋白轻链激酶(MLCK)和硫酸基转移酶1A3(SULT1A3)在子宫肌瘤组织中的表达及与子宫肌瘤发生发展的可能关系。方法:收集2019年1月-2021年6月在本院行手术切除治疗的子宫肌瘤患者60例子宫肌瘤组织和正常子宫组织,分别采用RT-PCR技术和免疫组化法测定MMP-2、MLCK、SULT1A3的mRNA及蛋白表达,分析各指标表达与子宫肌瘤发生的相关性。结果:子宫肌瘤组织MMP-2(1.61±0.23)、MLCK(11.87±1.60)的mRNA相对表达量均高于正常子宫组织(1.05±0.09、6.14±1.02),SULT1A3的mRNA相对表达量(0.69±0.11)低于正常子宫组织(1.65±0.44);子宫肌瘤组织中MMP-2蛋白阳性率(68.3%)、MLCK蛋白阳性率(76.7%)均高于正常子宫组织(25.0%、26.7%),子宫肌瘤组织SULT1A3蛋白阳性率(61.7%)低于正常子宫组织(100.0%)(均P<0.05)。mRNA表达,MMP-2与MLCK呈正相关,MMP-2、MLCK与SULT1A3均呈负相关性;蛋白表达,MMP-2与MLCK蛋白表达呈正相关,MMP-2、MLCK与SULT1A3蛋白表达均呈负相关性(均P<0.05)。结论:子宫肌瘤组织中MMP-2、MLCK呈高表达,SULT1A3呈低表达,且各指标具有相关性,这种异常表达可能与子宫肌瘤的发生有关。 展开更多
关键词 子宫肌瘤组织 基质金属蛋白酶-2 肌球蛋白轻链激酶 硫酸基转移酶1A3 相关性
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Study on the correlation of MLCK and FAP expression with uterine fibroid cell proliferation and invasion
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作者 Wei Lin Sha Ma Li Wang 《Journal of Hainan Medical University》 2017年第12期79-82,共4页
Objective:To study the correlation of myosin light chain kinase (MLCK) and fibroblast activation protein (FAP) expression with uterine fibroid cell proliferation and invasion. Methods:Uterine fibroids samples and norm... Objective:To study the correlation of myosin light chain kinase (MLCK) and fibroblast activation protein (FAP) expression with uterine fibroid cell proliferation and invasion. Methods:Uterine fibroids samples and normal uterine muscle samples next to fibroids that were surgically removed in Wuhan Red Cross Hospital between May 2014 and January 2017 were chosen, fluorescence quantitative PCR kits were used to deterct MLCK and FAP mRNA expression, and enzyme-linked immunosorbent assay kits were used to determine proliferation and invasion gene protein expression.Results: MLCK and FAP mRNA expression in uterine fibroids samples were significantly higher than those in normal uterine muscle samples, and Survivin, Livin, Bcl-2, Snail, N-cadherin and MMP2 protein expression were significantly higher than those in normal uterine muscle samples;Survivin, Livin, Bcl-2, Snail, N-cadherin and MMP2 protein expression in uterine fibroids samples with high MLCK and FAP expression were significantly higher than those in uterine fibroids samples with low MLCK and FAP expression.Conclusion: Highly expressed MLCK and FAP in uterine fibroids can promote the proliferation and invasion of uterine fibroids. 展开更多
关键词 UTERINE FIBROIDS myosin light chain kinase Fibroblast activation protein Proliferation INVASION
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Alterations in myosin heavy chain isoform gene expression during the transition from compensatory hypertrophy to congestive heart failure in rats 被引量:2
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作者 黄焰 刘惠灵 李云霞 《Chinese Medical Journal》 SCIE CAS CSCD 2001年第2期71-73,109-110,共5页
Objective To explore the molecular mechanism underlying the decreased velocity of tension rise in rat myocardium during congestive heart failure (CHF) and left ventricular hypertrophy (LVH) induced by aortic stenosis... Objective To explore the molecular mechanism underlying the decreased velocity of tension rise in rat myocardium during congestive heart failure (CHF) and left ventricular hypertrophy (LVH) induced by aortic stenosis.Methods The maximum velocity of tension rise (+dT/dtmax) was measured in left ventricular papillary muscle and the mRNA level of myosin heavy chain (MHC) isoforms in the left ventricle were detected by Northern blot analysis.Results The value of +dT/dtmax in CHF and LVH group were 64.17% and 37.15% lower than sham-operated controls (Sham) (P<0.01); values in the CHF group were 42.99% lower than that of LVH (P<0.01). The level of α-MHC mRNA in LVH was not different from that of the Sham (P>0.05), but decreased significantly in CHF to 42.3% of Sham and 56.1% of LVH (P<0.01). The level of β-MHC mRNA was up-regulated by 88.3% (P<0.01) in LVH compared with Sham and the level of β-MHC in CHF was 1.5-fold and 3.7-fold higher than that in LVH and Sham respectively (P<0.01). The ratio of α-MHC/β-MHC mRNA in LVH and CHF decreased to 42.4% and 9.8% respectively of the value in Sham (P<0.01). Correlation between α-MHC/β-MHC mRNA level and +dT/dtmax was analyzed which showed that these values were positively correlated with a correlation coefficient of 0.875 (P<0.01).Conclusion The decreased ratio of α-MHC/β-MHC mRNA was the major molecular mechanism underlying the decreased +dT/dtmax in CHF and LVH myocardium. The decreased ratio of α-MHC/β-MHC mRNA in LVH was mainly due to the up regulation of β-MHC mRNA while in CHF both down regulation of α-MHC and up regulation of β-MHC were involved. 展开更多
关键词 congestive heart failure · myosin heavy chain · left ventricular hypertrophy · animal model · atpase
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