Long noncoding RNAs HAND2-AS1 ultrasound microbubbles suppress hepatocellular carcinoma progression by regulating the miR-873-5p/tissue inhibitor of matrix metalloproteinase-2 axis

BACKGROUND Increasing data indicated that long noncoding RNAs(lncRNAs)were directly or indirectly involved in the occurrence and development of tumors,including hepatocellular carcinoma(HCC).Recent studies had found that the expression of lncRNA HAND2-AS1 was downregulated in HCC tissues,but its role in HCC progression is unclear.Ultrasound targeted microbubble destruction mediated gene transfection is a new method to overexpress genes.AIM To study the role of ultrasound microbubbles(UTMBs)mediated HAND2-AS1 in the progression of HCC,in order to provide a new reference for the treatment of HCC.METHODS In vitro,we transfected HAND2-AS1 siRNA into HepG2 cells by UTMBs,and detected cell proliferation,apoptosis,invasion and epithelial-mesenchymal transition(EMT)by cell counting kit-8 assay,flow cytometry,Transwell invasion assay and Western blotting,respectively.In addition,we transfected miR-837-5p mimic into UTMBs treated cells and observed the changes of cell behavior.Next,the UTMBs treated HepG2 cells were transfected together with miR-837-5p mimic and tissue inhibitor of matrix metalloproteinase-2(TIMP2)overexpression vector,and we detected cell proliferation,apoptosis,invasion and EMT.In vivo,we established a mouse model of subcutaneous transplantation of HepG2 cells and observed the effect of HAND2-AS1 silencing on tumor formation ability.RESULTS We found that UTMBs carrying HAND2-AS1 restricted cell proliferation,invasion,and EMT,encouraged apoptosis,and HAND2-AS1 silencing eliminated the effect of UTMBs.Additionally,miR-873-5p targets the gene HAND2-AS1,which also targets the 3’UTR of TIMP2.And miR-873-5p mimic counteracted the impact of HAND2-AS1.Further,miR-873-5p mimic solely or in combination with pcDNA-TIMP2 had been transformed into HepG2 cells exposed to UTMBs.We discovered that TIMP2 reversed the effect of miR-873-5p mimic caused by the blocked signalling cascade for matrix metalloproteinase(MMP)2/MMP9.In vivo results showed that HAND2-AS1 silencing significantly inhibited tumor formation in mice.CONCLUSION LncRNA HAND2-AS1 promotes TIMP2 expression by targeting miR-873-5p to inhibit HepG2 cell growth and delay HCC progression.

Expression of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-1 in hepatic stellate cells during rat hepatic fibrosis and its intervention by IL-10

AIM: To investigate the expression of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-1 in hepatic fibrosis and the antifibrogenic role of exogenous interleukin-10 (IL-10).METHODS: Hepatic fibrosis was induced by CCl4administration and 60 male Sprague-Dawley rats were randomly divided into normal control group (group N, 8rats), CCl4-induced group (group C, 28 rats) and IL-10-treated group (group I, 24 rats). At the beginning of the 7th and 11th wk, rats in each group were routinely perfused with pronase E and type Ⅳ collagenase through portal vein catheter and the suspension was centrifuged by 11%Nycodenz density gradient to isolate hepatic stellate cells (HSCs). RT-PCR was used to analyze mRNA of MMP-2 and TIMP-1 from freshly isolated cells. Densitometric data were standardized with β-actin signals. Immunocytochemistry was performed to detect MMP-2 and TIMP-1 expression in HSC cultured for 72 h.RESULTS: Compared to group N in the 7th wk, MMP-2and TIMP-1 mRNA increased in group C (P= 0.001/0.001)and group I (P = 0.001/0.009). The level of MMP-2 and TIMP-1 mRNA in group I was significantly lower than that in group C (P= 0.001/0.001). In the 11th wk, MMP-2 mRNAin group I was still lower than that in group C (P = 0.005),but both dropped compared with that in the 7th week (P = 0.001/0.004). TIMP-1 mRNA in group I was still lower than that in group C (P= 0.001), and increased in group C (P = 0.001) while decreased in group I (P = 0.042)compared with that in the 7th wk. Same results were found by immunocytochemistry.CONCLUSION: Expression of MMP-2 and TIMP-1 is increased in hepatic fibrosis. IL-10 exhibits an antifibrogenic effect by suppressing MMP-2 and TIMP-1 expression.

Correlation of matrix metalloproteinase－2, －9, tissue inhibitor－1 of matrix metalloproteinase and CD44 variant 6 in head and neck cancer metastasis

This study aimed to explore the molecular mechanism in tumor invasion and metastasis. The ex-pression of matrix metalloproteinase-2,-9 (MMP-2,MMP-9), tissue inhibitor-1 of matrix metalloprote inase(TIMP-1) , cell adhesion molecule 44 variant 6 (CD44v6) , HER2/neu and p53 was investigated in 154 pa-tients with head and neck squamous cell carcinoma (SCC) by ABC and ImmunoMax immunohistochemical method. Their clinical relevance and correlation were analysed. The expression of MMP-2, MMP-9, TIMP-1,CD44v6, HER2/neu and p53 was found in cancer cells in 87.01%, 85.71%, 68. 18%, 98.05%,55.19% and 50.65% cases respectively. Linear regression and correlation analysis revealed that there wasclose positive relationship ( P < 0.05) between the expression of MMP-2 and MMP-9, TIMP-1 and CD44v6,HER2/neu and MMP-9, MMP-2 and p53. Up-regulation of MMP-2 was accompanied by advanced T stage( P < 0.01 ) . There was also a trend of MMP-2 expression being related with tumor metastasis. Increased ex-pression of HER2/neu was found in patients with tumor recurrence( P < 0.05 ) . The expression of TIMP-1 washigher in laryngeal cancer than that in pharyngeal cancer, and higher in keratinizing and non-keratlnizing SCC than that in basaloid SCC ( P < 0.05 ) . These findings suggested that MMP-2 and MMP-9, HER2/neu andMMP-9, MMP-2 and p53 had a coordinate function in aggression of tumor; that MMP-2 had a more important function than MMP-9 in tumor invasion and metastasis; and that HER2/neu might serve as a biomarker forpoor prognosis in HNSCC.

Preparation and in vitro studies of microencapsulated cells releasing human tissue inhibitor of metalloproteinase-2

Objective: To prepare microencapsulated cells releasing human tissue inhibitor ofmetalloproteinase-2 (TIMP-2), and investigate their biological characteristics in vitro. Methods: Chinese hamster ovary (CHO) cells were stably transfected with a human TIMP-2 expression vector, encapsulated in barium alginate microcapsules and cultured in vitro. Morphological appearance of the microcapsules was observed under a light microscope. Cell viability was assessed using MTT (3-(4,5-dimethylthiazol2-yl)-2,5-diphenyltetrazolium bromide) assay. Enzyme linked immunosorbent assay (ELISA) and reverse zymography were used to confirm the release of biologically active TIMP-2 from the microcapsules. Cryopreservation study of the microencapsulated cells was carried out using dimethyl sulfoxide (DMSO) as preservative agent. Results: The microcapsules appeared like a sphere kept proliferating over the 6 weeks observed. No significant difference in TIMP-2 secretion was found between encapsulated and unencapsulated cells. Reverse zymography confirmed the bioactivity of MMP (matrix metalloproteinase) inhibition of TIMP-2.The cryopreservation process did not damage the microcapsule morphology nor the viability of the cells inside. Conclusion:Microencapsulated engineered CHO cells survive at least 6 weeks after preparation in vitro, and secrete bioactive TIMP-2 freely from the microcapsules.

重度烧伤患者入院时尿[TIMP-2]×[IGFBP7]水平与早期肾损伤的关系

目的 探讨重度烧伤患者入院时尿组织金属蛋白酶抑制剂-2[TIMP-2]×胰岛素样生长因子结合蛋白7[IGFBP7]水平与早期肾损伤的关系。方法 选取2018年11月—2022年6月本院收治的77例重度烧伤患者设为重度组、77例轻中度烧伤患者设为轻中度组,另选取同期77例体检正常者为正常组。比较3组尿[TIMP-2]×[IGFBP7]水平及血清胱抑素C(CysC)水平。根据重度烧伤患者在住院期间是否发生肾早期损伤分为非肾损伤组(45例)和肾损伤组(32例),比较非肾损伤组、肾损伤组重度烧伤患者尿[TIMP-2]×[IGFBP7]水平及血清CysC水平。分析入院时尿[TIMP-2]×[IGFBP7]、血清CysC对重度烧伤患者并发肾损伤的预测价值。结果 重度组烧伤患者尿[TIMP-2]×[IGFBP7]水平及血清CysC水平高于正常组、轻中度组(P<0.05),轻中度组烧伤患者尿[TIMP-2]×[IGFBP7]水平及血清CysC水平高于正常组(P<0.05)。肾损伤组重度烧伤患者尿[TIMP-2]×[IGFBP7]、血清CysC水平高于非肾损伤组(P<0.05)。入院时尿[TIMP-2]×[IGFBP7]、血清CysC预测重度烧伤患者并发肾损伤的AUC分别为0.873、0.699,截断值分别为1.06 [(ng/mL)^(2)/1000]、5.52 mmol/L,灵敏度分别为84.4%、59.4%,特异度分别为89.9%,73.3%。结论 发生早期肾损伤的重度烧伤患者入院时尿[TIMP-2]×[IGFBP7]水平及血清CysC水平较高,[TIMP-2]×[IGFBP7]水平或可成为预测重度烧伤患者并发肾损伤的潜在指标,为临床评估重度烧伤并发肾损伤提供参考。

前列腺突入膀胱程度及尿TIMP-2水平与前列腺增生患者膀胱出口梗阻严重程度的相关性分析

目的研究前列腺突入膀胱程度(Intravesical prostatic protrusion,IPP)及尿金属蛋白酶组织抑制剂-2(Tissue inhibitor of metalloproteinase-2,TIMP-2)与前列腺增生患者膀胱出口梗阻(Bladder outlet obstruction,BOO)严重程度的相关性。方法收集99例良性前列腺增生(Benign prostatic hyperplasia,BPH)患者纳入本研究,收集患者临床资料,检测患者IPP及尿TIMP-2,对患者进行尿动力学检测。根据国际前列腺症状评分(International prostate symptom score,IPSS)将患者分为3组,0~7评分为轻度组,共42例,8~19分为中度组,共25例,20~35分为重度组,共32例。采用Logistic回归分析3组患者的临床资料、IPP、尿TIMP-2、膀胱出口梗阻指数(Bladder outlet obstruction,BOOI)的相关性。采用受试者工作特征曲线(Receiver operating characteristic,ROC)分析IPP、TIMP-2检测预测BOO的敏感性。结果使用单因素方差分析法分析3组患者的临床指标,随着患病程度加重,年龄、TPV、IPSS、IPP、尿TIMP-2水平均有增加趋势,尿动力学指标中Qmax下降,Pdet.Qmax、BOOI、PVR均升高,且差异具有统计学意义(P均<0.05)。相关性分析结果显示,3组患者BOOI与年龄、BMI、TPV、PVR无显著相关性(P均>0.05)。轻度组、中度组和重度组患者BOOI与IPP、尿TIMP-2以及IPSS均呈正相关(P均<0.05)。ROC曲线分析显示IPP与尿TIMP-2单独预测BOO均具有较强敏感性,IPP联合尿TIMP-2检测敏感性更高(P均<0.05)。结论IPP、尿TIMP-2与前列腺增生患者BOO严重程度具有相关性,且IPP联合尿TIMP-2预测BOO具有较高敏感性。

Targeting epicardial adipose tissue:A potential therapeutic strategy for heart failure with preserved ejection fraction with type 2 diabetes mellitus

Heart failure with preserved ejection fraction(HFpEF)is a heterogeneous syndrome with various comorbidities,multiple cardiac and extracardiac pathophysiologic abnormalities,and diverse phenotypic presentations.Since HFpEF is a heterogeneous disease with different phenotypes,individualized treatment is required.HFpEF with type 2 diabetes mellitus(T2DM)represents a specific phenotype of HFpEF,with about 45%-50% of HFpEF patients suffering from T2DM.Systemic inflammation associated with dysregulated glucose metabolism is a critical pathological mechanism of HFpEF with T2DM,which is intimately related to the expansion and dysfunction(inflammation and hypermetabolic activity)of epicardial adipose tissue(EAT).EAT is well established as a very active endocrine organ that can regulate the pathophysiological processes of HFpEF with T2DM through the paracrine and endocrine mechanisms.Therefore,suppressing abnormal EAT expansion may be a promising therapeutic strategy for HFpEF with T2DM.Although there is no treatment specifically for EAT,lifestyle management,bariatric surgery,and some pharmaceutical interventions(anti-cytokine drugs,statins,proprotein convertase subtilisin/kexin type 9 inhibitors,metformin,glucagon-like peptide-1 receptor agonists,and especially sodium-glucose cotransporter-2 inhibitors)have been shown to attenuate the inflammatory response or expansion of EAT.Importantly,these treatments may be beneficial in improving the clinical symptoms or prognosis of patients with HFpEF.Accordingly,well-designed randomized controlled trials are needed to validate the efficacy of current therapies.In addition,more novel and effective therapies targeting EAT are needed in the future.

血清金属蛋白酶组织抑制剂3和性别决定区Y框蛋白2在2型糖尿病肾损伤早期诊断中的临床应用

目的探究血清金属蛋白酶组织抑制剂3(tissue inhibitors of metalloproteinases 3,TIMP3)和性别决定区Y框蛋白2(transcription factor determining region Y box protein 2,SOX2)对2型糖尿病(type 2 diabetes mellitus,T2DM)肾损伤的早期诊断。方法选取2022年4月至2023年4月在华北石油管理局总医院就诊的102例T2DM患者为研究对象,根据24 h尿蛋白排泄率(uri-nary albumin excretion rate,UAER)将T2DM患者分为肾损伤组(n=40)和无肾损伤组(n=62),另选取同期在华北石油管理局总医院行体检的健康者50名为对照组。酶联免疫吸附法测定所有受试者血清TIMP3、SOX2水平;比较3组受试者一般资料、糖化血红蛋白(hemoglobin A1c,HbA1c)、UAER、血肌酐(serum creatinine,Scr)、估算肾小球滤过率(estimated glomerular filtration rate,eGFR)、血清TIMP3、SOX2水平;Pearson相关性分析血清TIMP3与SOX2及两者与HbA1c、UAER、Scr、eGFR之间的关系;受试者工作特征曲线(receiver operating characteristic curve,ROC)分析血清TIMP3、SOX2对T2DM患者肾损伤的诊断价值。结果T2DM患者血清TIMP3[(0.68±0.17)μg/L比(1.35±0.35)μg/L]及eGFR[(105.99±20.56)mL·min^(-1)·(1.73 m^(2))比(133.15±26.18)mL·min^(-1)·(1.73 m^(2))^(-1)]显著低于对照组(P<0.05),且肾损伤患者血清TIMP3[(0.47±0.11)μg/L比(0.82±0.21)^(-1)μg/L]及eGFR[(74.69±10.22)mL·min^(-1)·(1.73 m^(2))比(126.18±27.23)mL·min^(-1)·(1.73 m^(2))^(-1)]显著低于无肾损伤患者(P<0.05);血清SOX2[(8.91±1.82)kU/L比(5.15±1.31)kU/L]及HbA1c[(8.80±1.55)%比(5.52±0.83)%]、UAER[(70.13±18.06)mg/24 h比(13.22±3.61)mg/24 h]、Scr[(82.14±15.23)µmol/L比(53.19±5.62)µmol/L]显著高于对照组(P<0.05),且肾损伤患者血清SOX2[(10.81±2.13)kU/L比(5.15±1.31)kU/L]及UAER[(156.83±40.29)mg/24 h比(13.22±3.61)mg/24 h]、Scr[(113.77±13.58)µmol/L比(53.19±5.62)µmol/L]显著高于无肾损伤患者(P<0.05)。Pearson相关性分析结果显示,TIMP3与UAER、Scr、HbA1c呈显著负相关(P<0.05),与eGFR呈显著正相关(P<0.05);SOX2与UAER、Scr、HbA1c呈显著正相关(P<0.05),与eGFR呈显著负相关(P<0.05);血清TIMP3与SOX2呈显著负相关(r=-0.590,P<0.05)。ROC结果显示,血清TIMP3联合SOX2诊断T2DM患者肾损伤的敏感度和特异度分别为95.0%、85.3%,显著高于TIMP3、SOX2单独测定。结论T2DM肾损伤患者血清TIMP3水平显著降低,SOX2水平显著升高,且两者与T2DM患者肾损伤密切相关,可用于T2DM患者肾损伤早期诊断。

Expression of Matrix Metalloproteinase and Its Tissue Inhibitor in Haemangioma

The action mechanism of matrix metalloproteinases-2 （MMP-2） and tissue inhibitor of metalloproteinases-2 （TIMP-2） in the genesis, development and degeneration of haemangioma was investigated by detecting their expression in the tissue of haemangioma in different phases by using the immunohistochemistry. Fifty paraffin-embedded specimens of skin capillary haemangioma were collected, which were documented in the Department of Pathology, Renmin Hospital of Wuhan University from 2000 to 2006. All samples were stained by regular HE method, and proliferative cell nuclear antigen （PCNA） was tested by immunohistochemical S-P method. The samples were classified according to the Mulliken criteria and the expression pattern of PCNA. Immunohistochemical S-P method was ap- plied to detect the expression of MMP-2 and TIMP-2 in proliferative and degenerative phases of cutaneous capillary haemangioma, and in normal skin tissues. In combination with the detection of the expression of factor Ⅷ-related antigen, it was verified that in haemangioma tissues, the cells expressing MMP-2 and TIMP-2 were vascular endothelial cells. The MMP-2 and TIMP-2 expression was quantitatively analyzed by image analysis system （HPIAS-1000）, and one-way ANOVA（107） and SNK（q） test were done to analyze average absorbance （A） and positive area rate of immunohistochemically positive particles by using SPSS11.5. The results showed： （1） Among 50 samples of haemangioma, there were 26 proliferative haemangiomas, and 24 degenerative haemangiomas, respectively; （2） The expression of MMP-2 was weak in normal vascular endothelial cells, cytoplasm of connective tissues and extracellular matrix around blood vessels. The expression of MMP-2 in proliferative group was significantly higher than in degenerative group and control group （normal skin） （P〈0.05）, but there was no statistically significant difference between the latter two groups; （3） TIMP-2 was highly expressed in normal tissues, degenerative vascular endothelial cells, cytoplasm of connective tissues and extracellular matrix around blood vessels. The expression level of TIMP-2 in proliferative phase was significantly lower than in degenerative phase （P〈0.05）, and the expression of TIMP-2 in proliferative phase was significantly different from that in degenerative phase and normal tissues （P〈0.05）. It was concluded that in proliferative phase of haemangioma, MMP-2 may promote over-proliferation of endothelial cells of haemangioma, and in degenerative phase, TIMP-2 can inhibit the proliferation of endothelial cells of haemangioma. The two substances play important roles in the genesis, development and degeneration of haemangiomas.

2型糖尿病合并冠心病患者TIMP-4、FFA水平和EAT厚度与颈部血管病变相关性分析

【目的】探讨2型糖尿病(T2DM)合并冠心病(CHD)患者组织型金属蛋白酶抑制剂4(TIMP-4)、游离脂肪酸(FFA)水平及心外膜脂肪组织(EAT)厚度与颈部血管病变相关性。【方法】选取2019年5月至2022年5月在本院行冠脉造影检查诊断为T2DM合并CHD的158例患者,根据是否存在颈部血管病变分为非颈部血管病变组(n=90)、颈部血管病变组(n=68)。收集并比较两组患者临床资料;采用酶联免疫吸附法(ELSA)检测两组患者血清TIMP-4、FFA水平;使用超声心动图测量EAT厚度;采用多因素Logistic回归分析T2DM合并CHD患者发生颈部血管病变的危险因素;采用受试者工作特征(ROC)曲线评估血清TIMP-4、FFA水平及EAT厚度对T2DM合并CHD患者发生颈部血管病变的诊断价值。【结果】与非颈部血管病变组比较,颈部血管病变组患者血清TIMP-4水平降低(P<0.05),FFA水平及EAT厚度升高(P<0.05);多因素Logistic回归分析显示:TIMP-4水平降低、FFA水平及EAT厚度升高是T2DM合并CHD颈部血管病变的危险因素(OR=3.968、2.587、4.125,均P<0.05);血清TIMP-4、FFA水平及EAT厚度联合诊断T2DM合并CHD患者发生颈部血管病变的AUC分别为0.753、0.506、0.856、0.915,灵敏度分别为83.10%、70.69%、83.1%、85.77%,特异度分别为85.60%、62.87%、89.6%、82.96%。【结论】血清TIMP-4、FFA水平及EAT厚度联合检测对T2DM合并CHD患者发生颈部血管病变有一定的诊断价值,可为临床早期诊治T2DM合并CHD颈部血管病变提供一定参考。

Imbalance of matrix metalloproteinase-9 and matrix metalloproteinase tissue inhibitor-1 may contribute to hemorrhage in cerebellar arteriovenous malformations

In this study, we determined the expression levels of matrix metalloproteinase-2 and -9 and matrix metalloproteinase tissue inhibitor-1 and -2 in brain tissues and blood plasma of patients undergoing surgery for cerebellar arteriovenous malformations or primary epilepsy （control group）. Immunohistochemistry and enzyme-linked immunosorbent assay revealed that the expression of matrix metalloproteinase-9 and matrix metalloproteinase tissue inhibitor-1 was significantly higher in patients with cerebellar arteriovenous malformations than in patients with primary epilepsy. The ratio of matrix metalloproteinase-9 to matrix metalloproteinase tissue inhibitor-1 was significantly higher in patients with hemorrhagic cerebellar arteriovenous malformations compared with those with non-hemorrhagic malformations. Matrix metalloproteinase-2 and matrix metalloproteinase tissue inhibitor-2 levels were not significantly changed. These findings indicate that an imbalance of matrix metalloproteinase-9 and matrix metalloproteinase tissue inhibitor-I, resulting in a relative overabundance of matrix metalloproteinase-9, might be the underlying mechanism of hemorrhage of cerebellar arteriovenous malformations.

Occurrence of Two Distinct types of Tissue Inhibitors of Metallo-proteinases-2 in Fugu rubripes

In this study, genes of two distinct tissue inhibitors of metalloproteinases-2 (TIMP-2) from Japanese puffer fishFugu rubripes, Fugu TIMP-2a and TIMP-2b, were cloned. The open reading frames of Fugu TIMP-2a and TIMP-2b cDNAsare composed of 660 and 657 nucleotides and 220 and 219 amino acids, respectively. Both Fugu TIMP-2s contain 12 cysteineresidues, which might form six disulfide bonds as in other animals’ TIMP-2s. Reverse-transcribed polymerase chain reactionanalysis showed the mRNAs of Fugu TIMP-2a and TIMP-2b to be expressed in some tissues examined with different expres-sion patterns. These findings suggest that the two distinct Fugu TIMP-2s might perform different functions in Fugu tissues.

miR-9-5p靶向TIMP2诱导多发性骨髓瘤细胞自噬和凋亡的机制

目的探究miR-9-5p和组织金属蛋白酶抑制因子2(TIMP2)相互作用对多发性骨髓瘤(MM)细胞自噬和凋亡的影响机制。方法采用实时荧光定量PCR(qRT-PCR)检测初诊MM和复发MM各9例患者骨髓样本中miR-9-5p和TIMP2的表达水平,分析两者表达水平的相关性。U266细胞分为miR-对照组、miR-9-5p组、pcDNA3.1组、pcDNA3.1-TIMP2组、miR-9-5p+pcDNA3.1组、miR-9-5p+pcDNA3.1-TIMP2组。采用流式细胞术、免疫荧光染色、蛋白质印迹实验检测过表达miR-9-5p和TIMP2对U266细胞自噬和凋亡的影响;双萤光素酶报告实验验证miR-9-5p和TIMP2的靶向关系。结果与初诊MM患者相比,复发MM患者miR-9-5p表达水平升高,TIMP2表达降低;miR-9-5p和TIMP2表达水平呈负相关(P<0.05)。与miR-对照组相比,miR-9-5p组MAP1LC3B-Ⅱ的表达水平降低,MAP1LC3B-Ⅰ和SQSTM1的表达水平增加,细胞凋亡率降低(P<0.05)。与pcDNA3.1组相比,pcDNA3.1-TIMP2组MAP1LC3B-Ⅱ的表达水平升高,MAP1LC3B-Ⅰ和SQSTM1的表达水平降低,细胞凋亡率增加(P<0.05)。生物信息学和双萤光素酶报告实验证实TIMP2是miR-9-5p的靶基因。结论miR-9-5p靶向TIMP2抑制MM细胞的自噬和凋亡,从而促进MM的发生发展。

血清TIMP-2、PLGF与高龄子痫前期患者胎儿生长受限的关系

目的 分析血清基质金属蛋白酶抑制因子(TIMP)-2、胎盘生长因子(PLGF)与高龄子痫前期(PE)患者发生胎儿生长受限(FGR)的关系。方法 采用前瞻性队列研究法选取2020年3月至2022年2月于新疆医科大学第一附属医院产检的138例高龄PE患者作为研究对象,纳入PE组;另选取同期在同一医院接受产检的138例高龄孕妇作为对照组。检查所有研究对象的肝肾功能[血肌酐(Scr)、谷丙转氨酶(ALT)、谷草转氨酶(AST)]、孕激素及血清TIMP-2、PLGF等。统计患者FGR发生情况,并分为FGR组和N-FGR组,采用Logistic回归分析血清TIMP-2、PLGF与高龄PE患者发生FGR的关系。结果 PE组血清TIMP-2水平高于对照组,血清PLGF水平低于对照组,差异均具有统计学意义(P<0.05)。138例高龄PE患者发生流产8例,胎停3例,退出研究2例,最终125例患者获得随访结果,胎儿娩出时确诊为FGR 27例。FGR组收缩压(SBP)、舒张压(DBP)、Scr及血清ALT、AST、TIMP-2水平均高于N-FGR组,血清PLGF水平低于N-FGR组,差异均具有统计学意义(P<0.05)。Logistic回归分析结果显示,血清TIMP-2高表达、PLGF低表达与高龄PE孕妇发生FGR有关(P<0.05)。结论 FGR患者血清TIMP-2显著升高,而PLGF水平显著降低;血清TIMP-2高表达、PLGF低表达是导致高龄PE孕妇FGR风险增加的危险因素。

红景天苷调控miR-20a-5p/TIMP2轴对类风湿关节炎成纤维样滑膜细胞增殖和迁移的影响

目的探讨红景天苷调控miR-20a-5p/金属蛋白酶组织抑制剂-2(TIMP2)轴对人类风湿关节炎成纤维样滑膜细胞(HFLS-RA)功能和活化的影响。方法以HFLS-RA细胞为研究对象,将HFLS-RA细胞分为肿瘤坏死因子-α(TNF-α)组、对照组、红景天苷组、miR-20a-5p抑制物阴性对照(inhibitor NC)组、miR-20a-5p抑制物组、红景天苷+miR-20a-5p模拟物阴性对照(mimic NC)组、红景天苷+miR-20a-5p模拟物组。实时荧光定量聚合酶链反应(qRT-PCR)检测HFLS-RA细胞中miR-20a-5p表达;酶联免疫吸附试验(ELISA)检测HFLS-RA细胞上清液中白细胞介素(IL)-1β、IL-6水平;细胞计数试剂盒8(CCK-8)法、5-乙炔基-2’-脱氧尿苷(EdU)染色检测HFLS-RA细胞增殖;划痕实验检测HFLS-RA细胞迁移;免疫印迹实验(Western blot)检测HFLS-RA细胞中TIMP2、细胞周期素D1(CyclinD1)、基质金属蛋白酶(MMP)-9蛋白表达;双荧光素酶验证miR-20a-5p与TIMP2的关系。结果与对照组比较,TNF-α组miR-20a-5p表达、IL-1β、IL-6水平、吸光度(OD_(450))值、EdU阳性细胞率、划痕愈合率及CyclinD1、MMP-9蛋白上调,TIMP2蛋白下调(P<0.05);与TNF-α组相比,红景天苷组miR-20a-5p表达、IL-1β、IL-6水平、OD_(450)值、EdU阳性细胞率、划痕愈合率及CyclinD1、MMP-9蛋白下调,TIMP2蛋白上调(P<0.05);与inhibitor NC组、TNF-α组相比比较,miR-20a-5p抑制物组miR-20a-5p表达、IL-1β、IL-6水平、OD_(450)值、EdU阳性细胞率、划痕愈合率及CyclinD1、MMP-9蛋白下调,TIMP2蛋白上调(P<0.05);与红景天苷+mimic NC组、红景天苷组相比,红景天苷+miR-20a-5p模拟物组miR-20a-5p表达、IL-1β、IL-6水平、OD_(450)值、EdU阳性细胞率、划痕愈合率及CyclinD1、MMP-9蛋白表达升高,TIMP2蛋白表达降低(P<0.05)。miR-20a-5p与TIMP2存在靶向调控关系。结论红景天苷可能通过调控miR-20a-5p/TIMP2抑制TNF-α诱导的HFLS-RA细胞增殖、迁移及炎症反应。

微小RNA-483-5p调控TIMP2表达对膀胱癌细胞增殖和侵袭的影响

目的探讨微小RNA-483-5p(miR-483-5p)对膀胱癌(BC)细胞增殖、迁移和侵袭的影响,并初步分析其可能的分子机制。方法通过TCGA数据库分析miR-483-5p在BC组织中的表达及与预后的关系,荧光实时定量PCR检测BC细胞T24中miR-483-5p和金属蛋白酶组织抑制因子(TIMP)-2的表达水平。将miR-483-5p模拟物(mimic)和阻碍物(inhibitor)转染T24细胞,应用CCK-8和Transwell法评估miR-483-5p对T24细胞增殖和迁移侵袭的影响。双荧光素酶报告基因实验分析miR-483-5p和TIMP-2间的靶向关系,挽救实验验证miR-483-5p的生物学功能是否通过TIMP-2发挥作用。结果与癌旁组织比较,BC组织中高表达miR-483-5p(P<0.05),高表达miR-483-5p者的总生存率低于低表达者(P<0.05)。miR-483-5p mimic可促进T24细胞的增殖、迁移和侵袭能力(P<0.05),miR-483-5p inhibitor则抑制T24细胞的增殖、迁移和侵袭能力(P<0.05)。TIMP2为miR-483-5p的下游靶点,干扰TIMP2可逆转miR-483-5p下调对T24细胞增殖、迁移和侵袭的抑制作用(P<0.05)。结论miR-483-5p在BC中高表达,与BC患者不良预后相关。miR-483-5p可下调TIMP2的表达来促进BC细胞的增殖、迁移和侵袭。

乙型肝炎肝硬化并发急性肾损伤患者血清和尿液NGAL、IGFBP7和TIMP-2水平变化及其临床意义探讨

目的 探讨乙型肝炎肝硬化并发急性肾损伤(AKI)患者血清和尿液中性粒细胞明胶酶原相关蛋白(NGAL)、胰岛素样生长因子结合蛋白7(IGFBP7)和金属蛋白酶组织抑制剂-2(TIMP-2)变化及其临床意义。方法 2020年3月~2022年12月我院收治的119例乙型肝炎肝硬化患者(并发AKI者38例,其中1期15例、2期14例和3期9例)和54例健康体检者,采用ELISA法检测血清NGAL水平及尿液NGAL、IGFBP7和TIMP-2水平。应用Cox单因素和多因素Logistic回归分析影响乙型肝炎肝硬化并发AKI的危险因素。结果 AKI组血清NGAL水平及尿液NGAL、尿液TIMP-2和IGFBP7水平分别为(371.7±60.2)μg/L、(59.7±7.3)μg/L、(3.3±0.6)ng/mL和(98.3±19.5)ng/mL,显著高于肝硬化组【分别为(82.3±15.7)μg/L、(10.7±2.5)μg/L、(2.4±0.5)ng/mL和(85.0±18.2)ng/mL,P<0.05】或健康人【分别为(46.5±10.9)μg/L、(7.9±1.2)μg/L、(0.7±0.1)ng/mL和(16.1±3.7)ng/mL,P<0.05】;AKI 3期患者血清NGAL水平及尿液NGAL、尿液TIMP-2和IGFBP7水平分别为(552.7±63.5)μg/L、(81.9±13.7)μg/L、(4.0±0.7)ng/mL和(110.4±15.1)ng/mL,显著高于1期患者【分别为(249.5±50.7)μg/L、(45.7±11.9)μg/L、(2.8±0.6)ng/mL和(90.3±10.9)ng/mL,P<0.05】或2期患者【分别为(386.3±59.8)μg/L、(60.4±9.5)μg/L、(3.4±0.7)ng/mL和(99.1±12.7)ng/mL,P<0.05】;本组并发AKI患者90 d生存率为57.9%;死亡组Child-Pugh C级、AKI 3期、肝性脑病、感染和上消化道出血患者占比显著高于生存组,血清NGAL水平及尿液NGAL、尿液TIMP-2和IGFBP7水平也显著高于生存组(P<0.05);多因素分析发现,Child-Pugh C级、AKI 3期、肝性脑病和上消化道出血及尿液NGAL和IGFBP7水平升高为乙型肝炎肝硬化并发AKI患者死亡的危险因素(P<0.05)。结论 乙型肝炎肝硬化并发AKI患者血清NGAL水平及尿液NGAL、TIMP-2和IGFBP7水平变化与病情严重程度有关,且尿液NGAL和IGFBP7水平显著升高为短期死亡的危险因素。

RECK、MMP-2、bcl-2蛋白在牙龈瘤患儿牙龈组织中的表达分析

目的 探讨RECK、基质金属蛋白酶-2(MMP-2)、B淋巴细胞瘤-2基因(bcl-2)蛋白在牙龈瘤患儿牙龈组织中的表达情况。方法 选取80例牙龈瘤患儿作为研究对象,所有患儿均行手术治疗,采集肿瘤组织及肿瘤旁健康牙龈组织,采用免疫组化SP法检测样本内RECK、MMP-2、bcl-2蛋白表达情况,比较肿瘤组织及健康牙龈组织中上述表达情况;并随访1年,分析RECK、MMP-2、bcl-2蛋白表达与其复发的关系。结果 肿瘤组织中RECK阳性率为36.25%,低于健康牙龈组织的77.50%,MMP-2阳性率、bcl-2蛋白阳性率分别为73.75%、67.50%,高于对照组的18.75%、22.50%,差异有统计学意义(P<0.05);80例患儿术后随访1年,共出现33例复发,复发率为41.25%(33/80);复发组RECK阳性率为15.15%,低于未复发组的51.06%,MMP-2阳性率、bcl-2蛋白阳性率为90.91%、93.94%,高于未复发组的61.70%、48.94%,差异有统计学意义(P<0.05)。结论 RECK、MMP-2、bcl-2蛋白在牙龈瘤患儿牙龈组织内存在不同表达,RECK阳性表达率低,MMP-2、bcl-2蛋白表达偏高,其表达与复发存在密切关系,可为完善早期治疗工作提供一定指导。