常染色体显性多囊肾病（ADPKD）是导致遗传性肾衰竭的主要病因之一，目前仍缺乏有效的手段来逆转其疾病进展。干细胞疗法因其在组织修复和免疫调节方面的潜力，成为研究的热点领域。本研究系统阐述了造血干细胞（HSCs）通过旁分泌作用、代谢重塑及免疫调控干预ADPKD病理进程的机制，并总结了其在急性肾损伤和慢性肾病中的临床转化证据。此外，文章比较分析了间充质干细胞（MSCs）和诱导多能干细胞（iPSCs）在ADPKD治疗中的独特优势与面临的挑战，为多模态干细胞疗法的开发提供理论支持。

    Autosomal dominant polycystic kidney disease（ADPKD）stands as a leading cause of inherited renal failure，with current therapeutic strategies lacking effective means to reverse disease progression．Stem cell therapy，owing to its potential in tissue repair and immunomodulation，has emerged as a focal point of research．This review systematically elucidates the mechanisms by which hematopoietic stem cells（HSCs）intervene in ADPKD pathology through paracrine effects，metabolic reprogramming，and immune regulation．Furthermore，it summarizes clinical translational evidence of HSCs in both acute kidney injury and chronic kidney disease．This article also comparatively analyzes the unique advantages and challenges of mesenchymal stem cells（MSCs）and induced pluripotent stem cells（iPSCs）in the context of ADPKD treatment，thereby providing theoretical support for the development of multimodal stem cell therapies．

       目的 探索α-突触核蛋白（α-Syn）干预对人单核细胞白血病细胞系（THP-1）巨噬细胞源性泡沫细胞的影响。方法 通过佛波酯（PMA）和氧化型低密度脂蛋白（ox-LDL）构建THP-1巨噬细胞源性泡沫细胞模型,使用不同浓度（33、66、100、133 nmol/L）α-Syn处理泡沫细胞,随后检测细胞胆固醇含量和炎症因子白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）及白细胞介素-8（IL-8）的mRNA表达以及核因子κB（NF-κB）和凝集素样氧化低密度脂蛋白受体-1（LOX-1）的蛋白表达变化。结果 高剂量（100和133 nmol/L）α-Syn处理可以减少THP-1巨噬细胞源性泡沫细胞内胆固醇的含量（P<0.05）,并且减少IL-1β、IL-6和IL-8的mRNA表达（P<0.05）。进一步发现（100 nmol/L和133 nmol/L）α-Syn可以降低THP-1巨噬细胞源性泡沫细胞p-NF-κB和LOX-1的蛋白表达（P<0.05）。结论 α-Syn可以降低THP-1源性巨噬细胞泡沫细胞胆固醇蓄积和炎症反应,可能是通过下调p-NF-κB和LOX-1蛋白表达。

      Objective To explore the effects of α-synuclein（α-Syn）intervention on human monocytic leukemia cell（THP-1）macrophage-derived foam cells．Methods The THP-1 macrophage-derived foam cell model was constructed by phorbol 12-myristate 13-acetate（PMA）and oxidized low-density lipoprotein（ox-LDL）．Foam cells were treated with different concentrations（33, 66, 100, and 133 nmol/L）of α-Syn, and the cellular cholesterol contents, as well as the mRNA expression of IL-1β、IL-6 and IL-8 were detected．Subsequently,alternation in protein expression of NF-κB and LOX-1 was measured．Results High-dose（100 and 133nmol/L）α-Syn treatment significantly reduced the levels of intracellular cholesterol in THP-1-derived macrophage foam cells（P<0.05）and decreased the mRNA expression of IL-1β、IL-6 and IL-8（P<0.05）．It was further found that（100 nmol/L and 133 nmol/L）α-Syn decreased the protein expression of p-NF-κB and LOX-1 in THP-1 macrophage-derived foam cells（P<0.05）．Conclusions The results of the present study suggest that α-Syn reduces cholesterol accumulation and inflammatory response in THP-1-derived macrophage foam cells, possibly by down-regulating p-NF-κB and LOX-1 protein expression．

目的 探讨微RNA-29b（miR-29b）对子宫内膜癌细胞增殖、迁移和侵袭的影响。方法 子宫内膜癌HEC-1-B细胞分为miR-29b模拟物组（MM组）、miR-29b阻遏物组（MR组）和阴性对照物组（MNC组）,分别转染miR-29b拟似物、miR-29b阻遏物和miR-29b阴性对照物,每组设置6个复孔。以实时定量逆转录PCR检测miR-29b表达,以水溶性四氮唑（WST-1）检测miR-29b对HEC-1-B子宫内膜癌细胞增殖的影响,以Transwell小室检测HEC-1-B子宫内膜癌细胞迁移和侵袭的影响,以Western blot法检测磷酸酶张力蛋白同源物（PTEN）-蛋白激酶 B（AKT）通路蛋白表达水平。结果 MNC组、MM组、MR组miR-29b相对表达量分别为（2 032.1±873.4）、（19 272.8±2 087.9）、（472.7±105.6）,组间比较差异有统计学意义（P<0.05）。MM组0、3、5、7 d时OD值分别为（0.32±0.06）、（0.53±0.08）、（1.13±0.12）和（1.92±0.14）,MNC组0、3、5、7 d时OD值分别为（0.34±0.09）、（0.71±0.08）、（1.67±0.21）和（3.49±0.24）,MR组0、3、5、7 d时OD值分别为（0.38±0.09）、（0.84±0.18）、（2.43±0.24）和（5.67±0.15）,3组0 d时OD值比较差异无统计学意义（P=0.216）,三组3 d、5 d、7 d时OD值比较差异存在统计学意义（P<0.001）。MNC组、MM组和MR组迁移细胞数分别为（403.9±23.8）（102.6±15.7）和（685.7±46.8）个,上述3组侵袭细胞数分别为（82.1±12.7）（38.2±10.6）和（124.6±21.6）个,MM组和MNC组上述指标比较差异均有统计学意义（P<0.05）,MR组和MM组上述指标比较差异均有统计学意义（P<0.05）。MNC组、MM组、MR组PTEN蛋白相对表达量分别为（0.25±0.08）、（0.69±0.11）、（0.11±0.05）,上述3组p-AKT蛋白相对表达量分别为（0.58±0.10）、（0.13±0.06）和（0.79±0.08）,上述3组AKT蛋白相对表达量分别为（0.38±0.09）、（0.37±0.11）和（0.37±0.08）,MM组与MNC组PTEN、p-AKT水平比较差异有统计学意义（P<0.05）,AKT水平比较差异无统计学意义（P>0.05）;MR组与MNC组、MM组PTEN、p-AKT水平比较差异有统计学意义（P<0.05）,AKT水平比较差异无统计学意义（P>0.05）。结论 过表达miR-29b对子宫内膜癌细胞增殖、迁移和侵袭具有抑制作用,靶向PTEN-AKT可能是其重要作用途径。

Objective To investigate the effects of microRNA-29b on proliferation,migration and invasion of endometrial cancer cells．Methods The endometrial cancer HEC-1-B cells were divided into micro29b mimetic group（MM group）,micro29b repressor group（MR group）and negative control group（MNC group）,and the micro29b mimetic,micro29b repressor and micro29b negative control were transfected into each group,six compound holes with each group．The real-time quantitative reverse transcription polymerase chain reaction（RT-PCR）was used to detect the expression of mi29b,WST-1 was used to detect the effect of mi29b on the proliferation of HEC-1-B endometrial cancer cells,Transwell chamber was used to detect the migration and invasion of HEC-1-B endometrial cancer cells,and Western blot was used to detect the expression level of PTEN-AKT pathway protein．Results The relative expression levels of microRNA-29b in MNC group,MM group and MR group were（2 032.1±873.4）,（19 272.8±2 087.9）and（472.7±105.6）,respectively,and there were significant differences between groups（P<0.05）．OD values of MM group at 0 d,3 d,5 d and 7 d were（0.32±0.06）,（0.53±0.08）,（1.13±0.12）and（1.92±0.14）respectively．The OD values of MNC group at 0,3,5 and 7 days were（0.34±0.09）,（0.71±0.08）,（1.67±0.21）and（3.49±0.24）respectively．The OD values of MR group at 0 d,3 d,5 d and 7 d were（0.38±0.09）,（0.84±0.18）,（2.43±0.24）and（5.67±0.15）respectively．There was no significant difference in OD value between the three groups on day 0 （P=0.216）．There were significant differences in OD value between the three groups on day 3,day 5 and day 7（P<0.001）．The number of migrating cells in MNC group,MM group and MR group were（403.9±23.8）cells,（102.6±15.7）cells and（685.7±46.8）cells,respectively．The number of invasive cells in the above three groups were（82.1±12.7）cells,（38.2±10.6）cells and（124.6±21.6）cells．There were significant differences in the above indexes between MM group and MNC group（P<0.05）,also between MR group and MM group（P<0.05）．The relative expression levels of PTEN protein in MNC group,MM group and MR group were（0.25±0.08）,（0.69±0.11）and（0.11±0.05）．The relative expression levels of p-AKT protein in the above three groups were（0.58±0.10）,（0.13±0.06）and（0.79±0.08）．The relative expression levels of AKT protein in the above three groups were（0.38±0.09）,（0.37±0.11）and（0.37±0.08）,respectively．Compared with MNC group,the levels of PTEN and p-AKT in MM group had statistical significance（P<0.05）,but there was no statistical difference in AKT level（P>0.05）．Compared with MNC group and MM group,the levels of PTEN and p-AKT in MR group had statistical significance（P<0.05）,and there was no statistical difference in AKT level（P>0.05）．Conclusions Overexpression of microRNA-29b can inhibit the proliferation,migration and invasion of endometrial cancer cells,and targeting PTEN-AKT may be an important pathway．

目的 研究核因子-κB激活剂1（Act1）对高糖诱导肾小管上皮细胞炎症反应及细胞外基质表达的调控作用。方法 培养肾小管上皮细胞MK2,设置对照组和高糖组,处理12 h、24 h、48 h、72 h后检测细胞中Act1的mRNA和蛋白表达水平;设置si-NC组（5.5 mmol/L葡萄糖培养基中转染NC siRNA）、si-NC+高糖组（30 mmol/L葡萄糖培养基中转染NC siRNA）、si-Act1+高糖组（30 mmol/L葡萄糖培养基中转染Act1 siRNA）,48 h后检测细胞中Act1、E-钙黏蛋白（E-cadherin）、N-钙黏蛋白（N-cadherin）、I型胶原（Col-I）、III型胶原（Col-III）的mRNA和蛋白表达水平以及培养基中肿瘤坏死因子-α（TNF-α）、白介素-1β（IL-1β）、干扰素-γ（IFN-γ）的含量。结果 高糖组处理12 h、24 h、48 h、72 h时细胞中Act1的mRNA和蛋白表达水平均高于对照组（P<0.05）;si-NC+高糖组处理48h时细胞中Act1、N-cadherin、Col-I、Col-III的mRNA和蛋白表达水平以及培养基中TNF-α、IL-1β、IFN-γ的含量均高于si-NC组,细胞中E-cadherin的mRNA和蛋白表达水平均低于si-NC组（P<0.05）;si-Act1+高糖组处理48h时细胞中Act1、N-cadherin、Col-I、Col-III的mRNA和蛋白表达水平以及培养基中TNF-α、IL-1β、IFN-γ的含量均低于si-NC+高糖组,细胞中E-cadherin的mRNA和蛋白表达水平均高于si-NC+高糖组（P<0.05）。结论 Act1表达增加对高糖诱导肾小管上皮细胞炎症反应激活和细胞外基质增多具有促进作用。

Objective To study the regulation effect of nuclear factors-κB activator 1（NF-κB activator 1,Act1）on high glucose induced inflammatory response and extracellular matrix expression in renal tubular epithelial cells．Methods Renal tubular epithelial cells MK2 were cultured and control group and high glucose group were set．The mRNA and protein expression of Act1 were detected after treatment for 12,24,48 and 72 hours．MK2 were divided into si-NC group（transfected with NC siRNA in 5.5 mmol/L glucose medium）,si-NC+high glucose group（transfected with NC siRNA in 30 mmol/L glucose medium）and si-Act1+high glucose group（transfected with Act1 siRNA in 30 mmol/L glucose medium）．The mRNA and protein expression of Act1,E-cadherin,N-cadherin,type I collagen（Col-I）,and type III collagen（Col-III）and the contents of tumor necrosis factor-α（TNF-α）,interleukin-1β（IL-1β）,interferon-γ（IFN-γ）in culture medium were detected．Results The mRNA and protein expression levels of Act1 in cells of high glucose group were higher than those of control group at 12 h,24 h,48 h and 72 h（P<0.05）．The mRNA and protein expression levels of Act1,N-cadherin,CoL-I,Col-III in cells and the contents of TNF-α,IL-1β,IFN-γ in culture medium of si-NC+high glucose group were higher than those of si-NC group,the mRNA and protein expression levels of E-cadherin in cells were lower than those of si-NC group at 48 h（P<0.05）．The mRNA and protein expression levels of Act1,N-cadherin,CoL-I,Col-III in cells and the contents of TNF-α,IL-1β,IFN-γ in culture medium of si-Act1+high glucose group were lower than those of si-NC+high glucose group,the mRNA and protein expression levels of E-cadherin in cells were higher than those of si-NC+high glucose group at 48 h（P<0.05）．Conclusions The increased expression of Act1 promotes the activation of inflammatory response and the increase of extracellular matrix in renal tubular epithelial cells induced by high glucose．

目的 采用体外试验的方法,研究弱酸性培养对人正常食管内皮细胞（HEEC）活力的影响及潜在的调控机制。方法 细胞培养液的pH值分别设为（6.0~6.5）和（7.0~7.4）。以中性培养组为对照。利用CCK8实验,检测弱酸培养条件下,不同时间点食管内皮细胞活力的变化。采用蛋白免疫印迹法检测p38和磷酸化p38（p-p38）的表达。利用酶联免疫吸附剂测定法检测培养基上清液中IL-1β和IL-8的表达水平,并检测加入p38激酶活性抑制剂SBS 203580后两者浓度的变化。结果 弱酸环境下,细胞活力下降。培养1 h时,弱酸组细胞活力为（96.4±8.0）%,培养3 h和6 h时分别为（88.7±6.2）%和（87.7±7.4）%。细胞中p38的水平与培养基的pH值无关。弱酸培养可以促使细胞内p-p38的含量增加。基线时,弱酸组p-p38的灰度值比值为（0.37±0.02）,在培养2 h和6 h时分别为（0.64±0.09）、（0.84±0.11）,差异显著（P<0.01）。弱酸刺激诱导食管内皮细胞表达更多的IL-8和IL-1β。基线时弱酸组上清液中IL-8和IL-1β的浓度分别为（8.64±1.31）pg/mL,（3.35±0.49）pg/mL。培养6 h后,二者的浓度分别上升至（36.85±2.02）pg/mL和（19.19±1.60）pg/mL,差异显著（P<0.01）。加入SBS 203580后,IL-8和IL-1β的浓度明显下降（P<0.05）。结论 弱酸刺激可以降低食管内皮细胞的活力。p38 MAPK可能通过调控IL-8和IL-1β的表达参与该调节过程。

Objective To study the effect of weak acidic culture on the viability of normal human esophageal endothelial cells(HEEC)and the potential regulatory mechanisms. Methods The pH values of cell culture medium were set at(6. 0-6. 5)and(7. 0-7. 4),respectively. The group in neutral medium was set as control. CCK8 experiment was used to detect the change of cell viability at different time points. The expressions of p38 and phosphorylated p38(p-p38)were detected by Western Blot experiment. Enzyme linked immunosorbent assay was used to detect IL-1β and IL-8 concentration in the medium supernatant before and after adding p38 activity inhibitor(SBS 203580). Results HEEC viability was decreased under weak acidic conditions. After 1 hour of cultivation,the HEEC viability was(96. 4±8. 0)%,after 3 and 6 hours,it decreased to(88. 7±6. 2)% and(87. 7±7. 4)%,respectively. The level of p38 in cells was independent of culture medium pH values. Weak acidic stimulation could promote an increase of p-p38 in HEEC. At baseline,the gray value ratio of p-p38 in the weak acidic group was(0. 37±0. 02),and after 2 and 6 hours of culturing,it increased to(0. 64±0. 09)and(0. 84±0. 11),respectively,which differences were significant(P<0. 01). More IL-8 and IL-1β were expressed after weak acidic stimulation. At baseline,the concentrations of IL-8 and IL-1β in the medium supernatant of weak acidic group were(8. 64±1. 31)pg/mL and(3. 35±0. 49)pg/mL. After 6 hours of culturing,they increased significantly to(36. 85±2. 02)pg/mL and(19. 19±1. 60)pg/mL(P<0. 01),while the concentrations were decreased after adding SBS 203580(P<0. 05). Conclusions The HEEC viability was reduced by weak acidic stimulation,p38 MAPK may participate in the process by regulating the expression of IL-8 and IL-1β.

目的 构建尺寸可变纳米递送系统PAMAM/DOX-pep并进行表征,检测其理化性质并评价其体外抗肿瘤效果与靶向性。方法 将阿霉素（DOX）物理包埋在阳离子聚合物PAMAM的疏水空腔内,以4-（N-马来酰亚胺基甲基）环己烷羧酸N-羟基琥珀酰亚胺酯（SMCC）作为交联剂,采用金属基质蛋白酶（MMP-2）敏感的多肽pep（CPLGVRGC）串联小粒径纳米颗粒形成大尺寸纳米递送系统（PAMAM/DOX-pep）,对各纳米颗粒的粒径、电位、理化性质以及对小鼠乳腺癌细胞（4T1）的抑制作用、细胞摄取效果和核靶向作用进行检测。结果 PAMAM/DOX粒径约为10 nm,载药率为23%,多肽pep交联后形成的PAMAM/DOX-pep粒径约为200 nm,可在低pH下缓释DOX,7天内体外保持稳定且溶血率低、安全无毒,其与MMP-2共孵育后细胞摄取量与核靶向性显著增加。结论 尺寸可变纳米颗粒有助于克服尺寸所引发的递送障碍,将药物靶向递送至乳腺癌细胞核内并发挥作用,为纳米递送系统的设计提供了新策略。

Objective To construct and characterize the size-variable nano-delivery system PAMAM/DOX-pep,examine its physicochemical properties and evaluate its antitumor and targeting effects in vitro. Methods Small particle size PAMAM/DOX was obtained by physically encapsulating DOX within the hydrophobic cavity of the cationic polymer PAMAM. The large size nano-delivery system(PAMAM/DOX-pep)was formed by tandem linking small size nanoparticles by MMP-2 sensitive peptide pep(CPLGVRGC)using SMCC as a cross-linker. The particle size,potential,physical and chemical properties,inhibitory effect,cell uptake and nuclear targeting effect of each nanoparticle on mouse breast cancer cells(4T1)were detected. Results The particle size of PAMAM/DOX was about 10 nm,and the drug loading rate was 23%. PAMAMAM/DOX-pep,formed after cross-linking of peptide,had a particle size of about 200 nm,which could release DOX slowly at low pH,and remained stable,safe and non-toxic in vitro for 7 days with low hemolysis rate,and its cellular uptake amount and nuclear targeting rate increased significantly after co-incubation with MMP. Conclusions Size-variable nanoparticles overcome size-induced delivery barriers to target and deliver drugs to the 4T1 nucleus,providing a new strategy for the design of nano delivery systems.

目的 探讨不同浓度的青藤碱对肾癌细胞增殖、细胞周期及凋亡的影响。方法 以不同浓度的青藤碱处理肾癌细胞786-O,采用四氮唑蓝盐法检测细胞增殖能力,流式细胞术检测细胞周期分布,Annexin-v FITC/PI双染流式细胞分析仪检测细胞凋亡率;采用实时荧光定量PCR及蛋白免疫印迹(WB)检测c-myc、Bax、Caspase-3等细胞周期、凋亡相关基因表达情况。结果 青藤碱显著抑制786-O细胞的增殖能力,诱导细胞周期G1/S期阻滞及细胞凋亡;且随着青藤碱浓度的增加,其抑制率也逐渐增加;青藤碱显著下调c-myc蛋白表达,而诱导凋亡蛋白Bax、Caspase-3表达上调。结论 青藤碱可以显著抑制786-O细胞中c-myc表达,使其增殖能力减弱,诱导细胞周期阻滞及凋亡。青藤碱可能具有潜在的抑制肾癌生长作用。

Objective To investigate the effects of different concentrations of sinomenine on proliferation, cell cycle and apoptosis of renal carcinoma cells. Methods The renal carcinoma cells were treated with different concentrations of sinomenine. MTS was used to analyze the effects of sinomenine on proliferation in 786-O renal carcinoma cells, the cell cycle changes were determined using flow cytometry, while the changes of apoptosis were detected by Annexin V-FITC / PI double staining. The expression of apoptosis-related proteins such as c-myc, Bax and Caspase-3 were detected by Western blot. Results Sinomenine significantly inhibited the proliferation of 786-O cells and induced cell cycle arrest and apoptosis in G1/S phase. With the increase of sinomenine concentration, the inhibition rate increased gradually. Sinomenine significantly down-regulated the expression of c-myc protein, while the expressions of the apoptotic protein Bax, Caspase-3 were up-regulated. Conclusions Sinomenine can significantly inhibit the expression of c-myc in 786-O cells, reduce proliferation ability, and induce cell cycle arrest and apoptosis. Sinomenine may have a potential therapeutic effect on renal cancer.

目的 通过回顾分析广州地区Rh阴性血的采供情况,了解Rh阴性红细胞的供应特点,更好地实施Rh阴性血液库存管理,保障输血安全。方法 统计2011—2017年广州血液中心Rh阴性血液采集量与供应量,血型及医院供血量的分布情况。结果 2011—2017年广州血液中心Rh阴性全血采集量为11 283 U,占总采血量的0.39%(11 283/2 902 485),供应总量为11 693 U,占红细胞类成分血总供应量的0.43%(11 693/2 762 229),其中悬浮红细胞供应比例逐年增加,冰冻解冻去甘油红细胞供应比例逐年下降,并从2014年起Rh阴性悬浮红细胞供应增长率明显高于血液采集增长率;4种血型供应量总体比较,差异有统计学意义(F=22.217,P＜0.05),A、B型Rh阴性悬浮红细胞供应增长最快,增长率分别为137.3%和134.1%,医院分布以三级以上及综合医院为主。结论 应根据广州地区Rh阴性红细胞采供特点,来持续改进Rh阴性血源招募和采供策略。通过建立一支Rh阴性定期献血者队伍,加强Rh阴性血液统筹管理和临床科室-输血科-采供血机构的沟通,促进Rh阴性血液的供需平衡,保障Rh阴性患者输血安全。

Objective To understand the characteristics of Rh-negative red blood cells (RBC) supply, improve the management of storage and guarantee the safety of blood transfusion. The collection and supply of Rh-negative RBC in Guangzhou city was retrospectively analyzed. Methods The amount of Rh-negative RBC collection and supply, the distribution of ABO blood type and the consumption of blood by different hospitals in Guangzhou Blood Center from 2011 to 2017 were statistically analyzed. Results During 2011-2017, 11 283 U Rh-negative RBC was collected in Guangzhou Blood Center, accounting for 0.39% of total RBC collection (11 283/2 902 485); 1 693 U Rh-negative RBC was supplied, accounting for 0.43% of the total blood supply (11 693/2 762 229). The proportion of suspended RBC supply increased year by year, in contrast with the decease of proportion of frozen defrosteddeglyceal RBC supply. In addition, the growth rate of Rh-negative suspended RBC supply was higher than that of RBC collection since 2014 (F=217, P<0.05). A-and B-Rh negative suspended RBC supply increased, with growth rates of 137.3% and 134.1%, respectively. The Rh negative RBC was predominantly supplied to Grade III and general hospitals. Conclusion By establishing a team of Rh-negative regular blood donors, the strategies for recruitment, collection and supply of Rh-negative RBC should be continuously improved according to the characteristics of Rh-negative RBC collection and supply in Guangzhou. The coordinating management of Rh-negative, as well as the communication of clinical departments, departments of transfusion and blood banks should be improved. In addition, the balance between supply and demand of Rh-negative blood should be promoted, and the safety of blood transfusion for Rh-negative patients can be improved.

目的 探讨miR-148a对大鼠急性胰腺炎细胞模型中细胞自噬的影响。方法 选取培养AR42J细胞,细胞分为4组,即正常对照组、模型组、miR-148a mimics组及miR-148a阴性对照组。利用Lipofectamine 2000转染miR-148a mimics及阴性对照miR-148a至AR42J细胞,继续培养48 h后,利用浓度为200 μmol的牛磺胆酸钠盐(TLCs)刺激以上两组及模型组AR42J细胞20 min,正常对照组不做处理,然后提取各组细胞蛋白及RNA。利用RT-qPCR检测各组细胞中miR-148a的表达;利用CCK8实验检测各组细胞的活性;利用ELISA法检测各组细胞培养液中炎性因子IL-6,IL-1β及TNF-α的含量;利用Western blot检测自噬相关的基因Beclin1、LC3Ⅰ、 LC3Ⅱ的表达。结果 RT-qPCR结果显示,与正常对照组相比较,模型组心肌细胞中miR-148a mRNA的表达降低,而miR-148a mimics组细胞中miR-148a mRNA的表达显著升高;CCK-8实验结果显示,转染miR-148a mimics至细胞后,可提高模型细胞的活性;ELISA实验结果显示,与模型组相比较,转染miR-148a mimics至细胞后,细胞培养液中炎性因子IL-6,IL-1β及TNF-α的含量显著降低;Western blot结果显示,与模型组相比较,转染miR-148a mimics至细胞后,可降低细胞中Beclin1的表达,降低LC3Ⅱ/LC3Ⅰ的比率。结论 利用miR-148a mimics提高TLCs刺激的细胞模型中的miR-148a表达后,细胞中Beclin1的表达降低,LC3Ⅱ/LC3Ⅰ的比率降低,抑制了细胞自噬,降低了炎性因子IL-6、IL-1β、TNF-α的释放,从而提高了细胞的活性,miR-148a可通过调节模型细胞的自噬而发挥细胞保护作用。

Objective To investigate the effect of miR-148a on autophagy in rat acute pancreatitis cell model. Methods AR42J cells were cultured and divided into 4 groups: normal control group, model group, miR-148a mimics group and miR-148a negative control group. miR-148a mimics and miR-148a negative control were transfected to AR42J cells with Lipofectamine 2 000, then cells were cultured for 48 h. The AR42J cells were stimulated with sodium taurocholate (TLCs) at a concentration of 200 μmol for 20 min, the normal control group was not treated, then the protein and RNA were extracted in each group. The expression of miR-148a was detected by RT-qPCR in each group. The activity of cells was detected by CCK8 assay in each group. The contents of IL-6, IL-1β and TNF-α in the cell culture medium were detected by ELISA. Western blot was used to detect the expression of autophagy related genes Beclin1, LC3Ⅰ and LC3Ⅱ. Results RT-qPCR results showed that the expression of miR-148a mRNA in model group was significantly lower than that in normal control group, while the expression of miR-148a mRNA in miR-148a mimics group was significantly higher than that in normal control group. The results of CCK-8 assay showed that miR-148a could significantly increase the activity of model cells stimulated by TLCs. The results of ELISA showed that the contents of IL-6, IL-1β and TNF-α in cell culture medium were significantly decreased after miR-148a mimics transfection, compared with the model group. Western blot showed that miR-148a mimics could significantly decrease the expression of Beclin1 and the ratio of LC3Ⅱ/LC3Ⅰ, compared with the model group. Conclusion miR-148a mimics was used to enhance the expression of miR-148a in cells model stimulated by TLCs, the expression of Beclin1 and the ratio of LC3Ⅱ/LC3Ⅰ were decreased, and the autophagy was inhibited. The release of IL-6, IL-1β and TNF-α was decreased and the activity of cells was increased. miR-148a plays a cellular protective role by regulating autophagy in model cells.

目的 探究超声-微泡介导的miR-128通过调节PTEN对乳腺癌细胞阿霉素耐药的影响。方法 qPCR检测miR-128在乳腺癌细胞系中的表达,并利用结合微泡的miR-128质粒(质粒+超声+SF6微泡)转染细胞,探究超声-微泡介导的miR-128对乳腺癌细胞阿霉素耐药的影响。CCK8实验检测乳腺癌细胞的活性;qPCR检测过表达miR-128后对PTEN的影响和对乳腺癌细胞阿霉素耐药的影响。结果 miR-128在阿霉素耐药乳腺癌细胞中低表达;过表达miR-128能够增加乳腺癌细胞对阿霉素的敏感性,超声-微泡介导的miR-128进一步增强了乳腺癌细胞对阿霉素的敏感性;miR-128通过调节PTEN从而促进乳腺癌细胞对阿霉素耐药。结论 miR-128过表达可以增强乳腺癌对阿霉素的敏感性,超声-微泡介导的miR-128进一步增强了乳腺癌细胞对阿霉素的敏感性,本研究为乳腺癌阿霉素耐药的治疗提供了新的分子靶标和治疗途径。

Objective To explore the effect of ultrasound-microbubble mediated miR-128 on doxorubicin resistance in breast cancer cells by regulating PTEN. Methods Quantitatine PCR (qPCR) was used to detect the expression of miR-128 in breast cancer cell lines, and the ultrasound-microbubble combined miR-128 plasmid(plasmid+ultrasound+SF6 microbubbles) was used to transfect the cells to explore the effects of ultrasound-microbubble mediated miR-128 on doxorubicin resistance in cancer cells. The CCK8 experiment was used to detect the activity of breast cancer cells; qPCR was used to detect the effect of overexpression of miR-128 on PTEN and the effect on doxorubicin resistance of breast cancer cells. Results miR-128 was under-expressed in doxorubicin-resistant breast cancer cells; overexpression of miR-128 increased the sensitivity of breast cancer cells to doxorubicin,ultrasound-microbubble mediated miR-128 further enhanced breast cancer cells sensitivity to doxorubicin; miR-128 promote resistance to doxorubicin in breast cancer cells by regulating PTEN. Conclusion Overexpression of miR-128 could enhance the sensitivity of breast cancer to doxorubicin. Ultrasound-microbubble mediated miR-128 further enhanced the sensitivity. This study provided a treatment for doxorubicin resistance in breast cancer with new molecular targets and therapeutic approaches.