MicroRNA-630 alleviates inflammatory reactions in rats with diabetic kidney disease by targeting toll-like receptor 4

BACKGROUND Diabetic kidney disease(DKD)is a major complication of diabetes mellitus.Renal tubular epithelial cell(TEC)damage,which is strongly associated with the inflammatory response and mesenchymal trans-differentiation,plays a significant role in DKD;However,the precise molecular mechanism is unknown.The recently identified microRNA-630(miR-630)has been hypothesized to be closely associated with cell migration,apoptosis,and autophagy.However,the association between miR-630 and DKD and the underlying mechanism remain unknown.AIM To investigate how miR-630 affects TEC injury and the inflammatory response in DKD rats.METHODS Streptozotocin was administered to six-week-old male rats to create a hypergly cemic diabetic model.In the second week of modeling,the rats were divided into control,DKD,negative control of lentivirus,and miR-630 overexpression groups.After 8 wk,urine and blood samples were collected for the kidney injury assays,and renal tissues were removed for further molecular assays.The target gene for miR-630 was predicted using bioinformatics,and the association between miR-630 and toll-like receptor 4(TLR4)was confirmed using in vitro investigations and double luciferase reporter gene assays.Overexpression of miR-630 in DKD rats led to changes in body weight,renal weight index,basic blood parameters and histopathological changes.RESULTS The expression level of miR-630 was reduced in the kidney tissue of rats with DKD(P<0.05).The miR-630 and TLR4 expressions in rat renal TECs(NRK-52E)were measured using quantitative reverse transcription polymerase chain reaction.The mRNA expression level of miR-630 was significantly lower in the high-glucose(HG)and HG+mimic negative control(NC)groups than in the normal glucose(NG)group(P<0.05).In contrast,the mRNA expression level of TLR4 was significantly higher in these groups(P<0.05).However,miR-630 mRNA expression increased and TLR4 mRNA expression significantly decreased in the HG+miR-630 mimic group than in the HG+mimic NC group(P<0.05).Furthermore,the levels of tumor necrosis factor-alpha(TNF-α),interleukin-1β(IL-1β),and IL-6 were significantly higher in the HG and HG+mimic NC groups than in NG group(P<0.05).However,the levels of these cytokines were significantly lower in the HG+miR-630 mimic group than in the HG+mimic NC group(P<0.05).Notably,changes in protein expression were observed.The HG and HG+mimic NC groups showed a significant decrease in E-cadherin protein expression,whereas TLR4,α-smooth muscle actin(SMA),and collagen IV protein expression increased(P<0.05).Conversely,the HG+miR-630 mimic group exhibited a significant increase in E-cadherin protein expression and a notable decrease in TLR4,α-SMA,and collagen IV protein expression than in the HG+mimic NC group(P<0.05).The miR-630 targets TLR4 gene expression.In vivo experiments demonstrated that DKD rats treated with miR-630 agomir exhibited significantly higher miR-630 mRNA expression than DKD rats injected with agomir NC.Additionally,rats treated with miR-630 agomir showed significant reductions in urinary albumin,blood glucose,TLR4,and proinflammatory markers(TNF-α,IL-1β,and IL-6)expression levels(P<0.05).Moreover,these rats exhibited fewer kidney lesions and reduced infiltration of inflammatory cells.CONCLUSION MiR-630 may inhibit the inflammatory reaction of DKD by targeting TLR4,and has a protective effect on DKD.

Association between polymorphisms in the Toll-like receptor 4,CD14,and CARD15/NOD2and inflammatory bowel disease in the Greek population

AIM: Crohn's disease(CD)and ulcerative colitis(UC)are multifactorial diseases with a significant genetic background.Apart from CARD15/NOD2 gene, evidence is accumulating that molecules related to the innate immune response such as CD14 or Toll-like receptor 4 (TLR4), are involved in their pathogenesis. In further exploring the genetic background of these diseases, we investigated the variations in the CARD15/NOD2 gene (Arg702Trp,Gly908Arg and Leu1007fsinsC), and polymorphisms in the TLR4 gene (Asp299Gly and Thr399Ile) as well as in the promoter of the CD14 gene (T/C at position -159) in Greek patients with CD and UC.METHODS: DNA was obtained from 120 patients with CD,85 with UC and 100 healthy individuals. Genotyping was performed by allele specific PCR or by PCR-RFLP analysis.RESULTS: The 299Gly allele frequency of the TLR4 gene and the T allele and TT genotype frequendes of the CD14 promoter were significantly higher in CD patients only compared to healthy individuals (P = 0.026＜0.05; P = 0.0048＜0.01 and P= 0.047＜0.05 respectively). Concerning the NOD2/CARD15mutations the overall presence in CD patients was significantly higher than that in UC patients or in controls.Additionally, 51.67% of the CD patients were carriers of a TLR4 and/or CD14 polymorphic allele and at least one variant of the NOD2/CARD15, compared to 27% of the UC patients. It should be pointed out that both frequencies significantly increased as compared with the 10% frequency of multiple carriers found in healthy controls. A possible interaction of the NOD2/CARD15 with TLR4 and especially CD14, increased the risk of developing inflammatory bowel disease (IBD).CONCLUSION: Our results indicate that co-existence of a mutation in either the TLR4 or CD14 gene, and in NOD2/CARD15is associated with an increased susceptibility to developing CD compared to UC, and to developing either CD or UC compared to healthy individuals.

Clinical significance of NOD2/CARD15 and Toll-like receptor 4 gene single nucleotide polymorphisms in inflammatory bowel disease

AIM： To evaluate the role of genetic factors in the pathogenesis of Crohn＇s disease （CD） and ulcerative colitis （UC）, we investigated the single nucleotide polymorphisms （SNPs） of NOD2/CARD15 （R702W, Gg08R and L1007finsC）, and Toll-like receptor 4 （TLR4） genes （D299G and T399I） in a selected inflammatory bowel disease （IBD） population coming from Southern Italy. METHODS： Allele and genotype frequencies of NOD2/ CARD15 （R702W, Gg08R and L1007finsC） and TLR4 （D299G and T399I） SNPs were examined in 133 CD patients, in 45 UC patients, and in 103 healthy controls. A genotype-phenotype correlation was performed. RESULTS： NOD2/CARD15 R702W mutation was significantly more frequent in CD （9.8%） than in controls （2.4%, P = 0.001） and in UC （2.3%, P = 0.03）. No significant difference was found between UC patients and control group （P 〉 0.05）. In CD and UC patients, no significant association with G908R variant was found. L1007finsC SNP showed an association with CD （9.8%） compared with controls （2.9%, P = 0.002） and UC patients （2.3%, P = 0.01）. Moreover, in CD patients, G908R and L1007finsC mutations were significantly associated with different phenotypes compared to CD wild-type patients. No association of IBD with the TLR4 SNPs was found in either cohort （allele frequencies： D299G-controls 3.9%, CD 3.7%, UC 3.4%, P 〉 0.05; T399I-controls 2.9%, CD 3.0%, UC 3.4%, P 〉 0.05）. CONCLUSION： These findings confirm that, in our IBD patients selected from Southern Italy, the NOD2/ CARD15, but not TLR4 SNPs, are associated with increased risk of CD.

The changes of negative-regulatory factors A20,IRF-4 and TRAF4 of toll-like receptor signal pathways in immunological pathogenesis of Kawasaki disease

To investigate the role of negative-regulatory factors A20, IRF-d and TRAFd of the toll-like receptor （TLR） signal pathways in immunological pathogenesis of Kawasaki disease （KD）, 48 children with Kawasaki disease, 16 children with infectious disease （ID） and 16 age-matched healthy children were studied. Reverse-transcription PCR （RT-PCR） and real-time PCR were used to evaluate the expression levels of negative-regulatory and effective factors in toll-like receptor d （TLRd） signal pathways and proinflammatory factors in peripheral blood monocyte/macrophage （MC）. In this study, expression levels of TLRd, MD-2, MyD88, IRAK-d, TRAF6, TAK1, TAB1 and TAB2 mRNA in KD group were detected to be elevated significantly during acute phase of KD. Transcription levels of negative-regulatory factors A20, IRF-4 and TRAFd mRNA in KD or ID patients increased remarkably. However, expressions of IRF-4 and TRAFd in KD patients were detected to be lower than that in ID patients, except that transcription levels of A20 were found to be higher than that in ID patients. Simultaneously, expressions of proinflammatory cytokines such as L-1β, IL-6 and TNF-α in KD patients were significantly elevated compared with those in ID patients. Furthermore, it was found that stimulation of lipopolysaccharide （LPS） remarkably up-regulated the expressions of negative-regulatory factors A20, IRF-d and TRAFd in KD patients or healthy volunteers. The mRNA levels of all the three factors in KD patients were found to be lower than that in the latter. In addition, transcription levels of IRF-d and TRAFd in KD patients with coronary artery lesion （KD-CAL^＋ ） were detected to be lower than those in KD patients without coronary artery lesion （KD-CAL-） during acute phase, while that of A20 in KD-CAL^＋ group were lower than that in the latter. And the levels of expressions of proinflammatory cytokines in KD-CAL^＋ group were found to be higher than those in KD-CAL^- group （P 〈 0.01）. These findings suggest that aberrant expression of negative-regulatory factors of TLRs signal pathways may be involved in immunological pathogenesis of Kawasaki disease.

Metformin regulates inflammation and fibrosis in diabetic kidney disease through TNC/TLR4/NF-κB/miR-155-5p inflammatory loop

BACKGROUND Type 2 diabetes mellitus(T2DM)is significantly increasing worldwide,and the incidence of its complications is also on the rise.One of the main complications of T2DM is diabetic kidney disease(DKD).The glomerular filtration rate(GFR)and urinary albumin creatinine ratio(UACR)increase in the early stage.As the disease progresses,UACR continue to rise and GFR begins to decline until endstage renal disease appears.At the same time,DKD will also increase the incidence and mortality of cardiovascular and cerebrovascular diseases.At present,the pathogenesis of DKD is not very clear.Therefore,exploration of the pathogenesis of DKD to find a treatment approach,so as to delay the development of DKD,is essential to improve the prognosis of DKD.AIM To detect the expression of tenascin-C(TNC)in the serum of T2DM patients,observe the content of TNC in the glomerulus of DKD rats,and detect the expression of TNC on inflammatory and fibrotic factors in rat mesangial cells(RMCs)cultured under high glucose condition,in order to explore the specific molecular mechanism of TNC in DKD and bring a new direction for the treatment of DKD.METHODS The expression level of TNC in the serum of diabetic patients was detected by enzyme-linked immunosorbent assay(ELISA),the protein expression level of TNC in the glomerular area of DKD rats was detected by immunohistochemistry,and the expression level of TNC in the rat serum was detected by ELISA.Rat glomerular mesangial cells were cultured.Following high glucose stimulation,the expression levels of related proteins and mRNA were detected by Western blot and polymerase chain reaction,respectively.RESULTS ELISA results revealed an increase in the serum TNC level in patients with T2DM.Increasing UACR and hypertension significantly increased the expression of TNC(P<0.05).TNC expression was positively correlated with glycosylated haemoglobin(HbA1c)level,body mass index,systolic blood pressure,and UACR(P<0.05).Immunohistochemical staining showed that TNC expression in the glomeruli of rats with streptozotocin-induced diabetes was significantly increased compared with normal controls(P<0.05).Compared with normal rats,serum level of TNC in diabetic rats was significantly increased(P<0.05),which was positively correlated with urea nitrogen and urinary creatinine(P<0.05).The levels of TNC,Toll-like receptor-4(TLR4),phosphorylated nuclear factor-κB p65 protein(Ser536)(p-NF-κB p65),and miR-155-5p were increased in RMCs treated with high glucose(P<0.05).The level of TNC protein peaked 24 h after high glucose stimulation(P<0.05).After TNC knockdown,the levels of TLR4,p-NF-κB p65,miR-155-5p,connective tissue growth factor(CTGF),and fibronectin(FN)were decreased,revealing that TNC regulated miR-155-5p expression through the TLR4/NF-κB p65 pathway,thereby regulating inflammation(NF-κB p65)and fibrosis(CTGF and FN)in individuals with DKD.In addition,metformin treatment may relive the processes of inflammation and fibrosis in individuals with DKD by reducing the levels of the TNC,p-NF-κB p65,CTGF,and FN proteins.CONCLUSION TNC can promote the occurrence and development of DKD.Interfering with the TNC/TLR4/NF-κB p65/miR-155-5p pathway may become a new target for DKD treatment.

Analysis of TLR4 and TLR2 polymorphisms in inflammatory bowel disease in a Guangxi Zhuang population

AIM: To study the polymorphisms of toll-like receptor 4 (TLR4) gene Asp299Gly, Thr399Ile and TLR2 gene Arg753Gln, Arg677Trp and susceptibility to inflammatory bowel disease (IBD) in the Zhuang population from Guangxi, China. METHODS: A case-control study was performed from February 2007 to October 2011 which included 146 Zhuang patients with IBD in the experimental group and 164 healthy Zhuang subjects who acted as the control group. All patients and healthy subjects were from the Guangxi Zhuang Autonomous Region of China. Genomic DNA was extracted from intestinal tissue by the phenol chloroform method. TLR4 gene Asp299Gly, Thr399Ile and TLR2 gene Arg753Gln, Arg677Trp were amplified by polymerase chain reaction (PCR), and then detected by PCR-restriction fragment length polymorphism (RFLP). RESULTS: The TLR4 gene Asp299Gly was digested using Nco Ⅰ restriction enzyme, and a single band of 249 bp was observed which showed that it was a wild type (AA). The TLR4 gene Thr399Ile was digested using Hinf Ⅰrestriction enzyme and only the wild type (CC) was detected. In addition, the TLR2 gene Arg-677Trp was digested using Aci Ⅰ restriction enzyme and only the wild type (CC) was detected. The TLR2 gene Arg753Gln was digested using Pst Ⅰ restriction enzyme. Only the wild type (GG) as a single band of 254 bp was observed during RFLP. Overall, no heterozygous or homozygous single nucleotide polymorphism mutations were found in patients with Crohn's disease and ulcerative colitis both in the TLR4 gene Asp299Gly, Thr399Ile and the TLR2 gene Arg677Trp, Arg753Gln in the Zhuang population from the Guangxi Zhuang Autonomous Region of China. CONCLUSION: The TLR4 gene Asp299Gly, Thr399Ile and TLR2 gene Arg753Gln, Arg677Trp polymorphisms may not be associated with IBD in the Zhuang population from the Guangxi Zhuang Autonomous Region of China.

MicroRNA-630:A potential guardian against inflammation in diabetic kidney disease

In this editorial,we comment on the article by Wu et al published“MicroRNA-630 alleviates inflammatory reactions in rats with diabetic kidney disease by targeting toll-like receptor 4”.Diabetic kidney disease(DKD)stands as a significant complication occurring from diabetes mellitus,which contributes substantially to the morbidity and mortality rates worldwide.Renal tubular epithelial cell damage,often accompanied by inflammatory responses and mesenchymal transdifferentiation,plays a pivotal role in the progression of DKD.Despite extensive research,the intricate molecular mechanisms underlying these processes remain to be determined.Wu et al remarkable work identifies microRNA-630(miR-630)as an emerging potential regulator of cell migration,apoptosis,and autophagy,prompting investigation into its association with DKD pathogenesis.This study endeavors to elucidate the impact of miR-630 on TEC injury and the inflammatory response in DKD rats.The role of miR-630 in human DKD will be of interest for future studies.

MicroRNA-630:A promising avenue for alleviating inflammation in diabetic kidney disease

Diabetic kidney disease(DKD)is one of the complications of diabetes,affecting millions of people worldwide.The relentless progression of this condition can lead to kidney failure,requiring life-altering interventions such as dialysis or transplants.Accumulating evidence suggests that immunologic and inflammatory elements play an important role in initiating and perpetuating the damage inflicted on renal tissues,exacerbating the decline in organ function.Toll-like receptors(TLRs)are a family of receptors that play a role in the activation of the innate immune system by the recognition of pathogen-associated molecular patterns.Recent data from in vitro and in vivo studies have highlighted the critical role of TLRs,mainly TLR2 and TLR4,in the pathogenesis of DKD.In the diabetic milieu,these TLRs recognize diabetic-associated molecular signals,triggering a proinflammatory cascade that initiates and perpetuates inflammation and fibrogenesis in the diabetic kidney.Emerging non-traditional strategies targeting TLR signaling with potential therapeutic implications in DKD have been proposed.One of these approaches is the use of microRNAs,small non-coding RNAs that can regulate gene expression.This editorial comments on the results of this approach carried out in a rat model of diabetes by Wu et al,published in this issue of the World Journal of Diabetes.The results of the experimental study by Wu et al shows that microRNA-630 decreased levels compared to non-diabetic rats.Additionally,microRNA-630 exerted anti-inflammatory effects in the kidneys of diabetic rats through the modulation of TLR4.These findings indicate that the microRNA-630/TLR4 axis might represent a pathological mechanism of DKD and a potential therapeutic target capable of curbing the destructive inflammation characteristic of DKD.

Haoqin Qingdan Decoction(蒿芩清胆汤)and Ribavirin Therapy Downregulate CD14 and Toll-Like Receptor 4 in Febrile Disease with Dampness-Heat Syndrome in A Mouse Model

Objective： To evaluate the effect of Chinese medicine Haoqin Qingdan Decoction （蒿芩清胆汤, HQD） for febrile disease dampness-heat syndrome （FDDHS）. Methods： Forty mice were divided into four groups, including normal control, FDDHS （induced by Radix et Rhizoma Rhei recipe and influenza virus A1 FM1 model）, HQD, and the ribavirin groups （10 in each）. The normal control and FDDHS groups were administered normal saline. HQD and the ribavirin groups were administered HQD and ribavirin intragastrically once daily at a dose of 64 g/（kg.d） and 0.07 g/（kg.d）, respectively for 7 days. Lethargy, rough hair, diarrhea, tongue color and sole color were evaluated for pathological changes in morphology. The tongue and lung tissues were collected for histology. The CD14 and toll-like receptor 4 （TLR4） expression levels were measured using real-time quantitative polymerase chain reaction. Results： More than 80% of the FDDHS mice showed hypokinesia and lethargy, and pathological changes associated with rough hair, diarrhea, tongue color and sole color. With advanced treatment for 7 days, the thick greasy tongue fur of the HQD and ribavirin groups were thinner than that of the FDDHS group （P〈0.05）, and it was the thinnest in the ribavirin group as compared with that in other groups （P〈0.05）. The CD14 and TLR4 expression levels in the lung tissues of HQD and ribavirin groups significantly delined compared with the model group （P〈0.05 or P〈0.01）. CD14 was down-regulated more remarkably in the HQD group compared with the ribavirin group （P〈0.05）, whereas the converse was true with TLR4 （P〈0.05）. Conclusions： We established a FDDHS mouse model showing systemic clinical symptoms. Both HQD and ribavirin can inhibit the expression of CD14 and TLR4 in FDDHS mice, while the effect of ribavirin might be much more violent. The expression changes of CD14 and TLR4 consistently refers to lipopolysaccharide, the commonly and hotly inducing factor in FDDHS.

Toll样受体4信号通路在心力衰竭中的应用价值

心力衰竭是全球发病率最高的疾病之一,在心力衰竭及其合并症的发生和发展过程中炎症机制起着至关重要的作用。大量研究表明Toll样受体4信号通路可通过介导不同的炎症因子,对心肌造成损害。因此近期发现Toll样受体4信号通路可用于预测心力衰竭及其合并症病情的严重程度及预后,同时也可作为许多新型抗心力衰竭药物的靶点,改善症状,使心力衰竭患者获益。现就Toll样受体4信号通路在心力衰竭中上述两方面的应用价值做一综述。

Mechanism of action of verbascoside in mice with Parkinson's disease based on molecular docking,molecular dynamics and in vivo experiments

Objective:To explore the improving effect of verbascoside on Parkinson's disease(PD)model mice and to clarify the possible mechanism.Methods:60 C57BL/6 male mice were randomly divided into normal group,model group,low,middle and high dose groups(30,60 and 120 mg/kg,respectively).After a week of adaptive culture,except for the blank group,the subacute model of PD was established by intraperitoneal injection of 1-methyl-4-phenyl-1(MPTP)in each group for 5 consecutive days.After a week of adaptive culture,except for the blank group,the subacute model of PD was established by intraperitoneal injection of 1-methyl-4-phenyl-1(MPTP)in each group for 5 consecutive days.The establishment of the model,the drug group was intragastrically administered continuously for 7 d,and the mice in the normal group were injected with the same amount of saline.The behavioral changes of mice were observed by open field test,pole climbing test and grip test;the morphology,apoptosis and Nissl bodies of substantia nigra neurons were observed by HE staining and Nissl staining;the ultrastructural changes of neurons were observed by transmission electron microscope;the positive expressions of tyrosine hydroxylase(TH),α-synuclein(α-Syn)and toll-like receptor 4(TLR4)were detected by immunohistochemistry.The protein levels of TH,α-Syn,TLR4,NF-κB and P-NF-κB in substantia nigra of mice were detected by Western blot,and the levels of inflammatory factors IL-1β,IL-6 and tumor necrosis factor-α(TNF-α)in serum were detected by Elisa.Molecular docking was used to verify the binding ability of verbascoside to TLR4/NF-κB signal pathway and related factors,and molecular dynamics was used to verify the binding mode and stability of verbascoside with compounds.Results:compared with the normal group,the total walking distance,average speed,free activity time,pole climbing time and forelimb grip in the model group were significantly longer than those in the normal group(P<0.05).The ultrastructural changes of neurons,nuclear lysis,deformation and mitochondrial damage were observed under transmission electron microscope,and the number and morphological changes of HE staining substantia nigra neurons were decreased(P<0.05).Nissl staining showed that the number of Nissl positive cells decreased(P<0.05),the positive expression of TH decreased and the positive expression of α-syn and TLR4 increased(P<0.05).Western blotting showed that the expression of TH protein decreased,the expression of α-syn,TLR4,NF-κB p65 protein increased(P<0.05),and the expression of IL-1β,IL-6 and TNF-α in serum increased significantly(P<0.05).Compared with the model group,the activity ability of mice in the treatment group was significantly improved(P<0.05),the morphology of neurons gradually recovered and the number of neurons increased(P<0.05).The number of Nissl positive cells increased(P<0.05).The expression of TH protein increased,while the expression of α-syn,TLR4,NF-κBp65 protein decreased(P<0.05),and the expression of IL-1β,IL-6,TNF-α in serum decreased significantly(P<0.05).The results of molecular docking and molecular dynamics simulation show that the combination of mulberry glycoside with the compound is good and the property is stable.Conclusion:verbascoside has protective effect on PD mice,and its mechanism may be related to TLR4/NF-κB pathway,and then regulate neuroimmunity.

GDF-15、VEGF、CXCR4与冠心病PCI术后支架再狭窄的关系

目的 探讨生长分化因子-15(GDF-15)、血管内皮生长因子(VEGF)、CXC族趋化因子受体4(CXCR4)水平与冠心病患者经皮冠状动脉介入术(PCI)术后支架再狭窄(ISR)的关系。方法 选取2021年3月~2022年6月河南省荣军医院收治的118例冠心病患者,均接受PCI,根据PCI术后是否发生ISR分为ISR组(n=24例)和非ISR组(n=94例),采用酶联免疫吸附法测定GDF-15、CXCR4,双抗体夹心-酶联免疫吸附法测定VEGF,分析血清GDF-15、VEGF、CXCR4与冠心病患者PCI术后ISR的相关性及预测价值。结果 ISR组冠脉病变支数、植入支架数量多于非ISR组,冠脉病变长度长于非ISR组,术前Gensini积分、左室射血分数(LVEF)、D-二聚体(D-D)、纤维蛋白原(FIB)、GDF-15、VEGF高于非ISR组,CXCR4低于非ISR组,差异具有统计学意义(P<0.05)。FIB、GDF-15、VEGF水平升高和CXCR4水平下降是PCI术后发生ISR的独立危险因素,差异具有统计学意义(P<0.05);进一步校正FIB影响后,GDF-15、VEGF、CXCR4仍与PCI术后ISR发生有关。血清GDF-15、VEGF、CXCR4联合预测PCI术后ISR的AUC明显大于各指标单一预测,差异具有统计学意义(P<0.05)。结论 血清GDF-15、VEGF、CXCR4水平与冠心病患者PCI术后ISR具有显著相关性,三者均对ISR具有预测价值,联合检测能为临床提供更为可靠的数据支持。

磷酸二酯酶4在心脏疾病中作用的研究进展

磷酸二酯酶4(PDE4)是细胞内特异性cAMP水解酶,PDE4可以通过调节心肌细胞内的cAMP水平,进而影响cAMP参与的一系列生理功能的调节。目前,PDE4在慢性阻塞性肺病和银屑癣等疾病中作用的研究较为广泛,该综述旨在介绍PDE4在实验动物和人心肌肥大、心衰和心律失常中作用的研究进展。首先,通过观察大鼠心肌肥大模型发现PDE4水平有明显降低,进而对人类病变的心脏进行研究,表明患病的心脏上PDE4A和PDE4D水解活性也有明显下降;其次,在人心衰的心肌细胞上,存在于心脏兰诺定2型受体/Ca2+复合体中的PDE4D3,水解cAMP的活性也有显著降低;最后,在β肾上腺素刺激下,离体人心肌细胞中PDE4受到抑制后,细胞内cAMP水平和L型Ca2+电流均增加,表明PDE4受抑制会增加心率失常的发生率。

冠心病患者血小板Toll样受体4的表达及其意义

目的观察冠状动脉粥样硬化性心脏病(冠心病)患者血小板Toll样受体4(TLR4)的表达及其与血小板P-选择素、血浆TNF-α水平的关系,探讨血小板参与冠心病发病的可能机制。方法冠心病病人30例,健康对照者10例,流式细胞术检测血小板TLR4和血小板P-选择素的表达,ELISA方法检测血浆TNF-α水平;另用Westernblot检测血小板总TLR4。结果血小板TLR4、血小板P-选择素表达和血浆TNF-α水平在冠心病组分别是26.35±1.79%、30.62±3.22%和15.61±1.36pg/ml;健康对照组分别是18.78±2.15%、14.50±1.13%和8.79±1.20pg/ml,前者明显高于后者(P<0.05)。变量间相关分析显示,血小板TLR4的表达水平与P-选择素及血浆TNF-α呈正相关(Pearson相关系数分别为0.618和0.745,P<0.01)。结论冠心病患者血小板TLR4表达增高,可能与冠心病血小板的高反应性和发病有关。

血清miR-223、CXCR4与冠状动脉支架内再狭窄的关系探讨

目的分析血清微小RNA-223(miR-223)、CXC族趋化因子受体4(CXCR4)表达水平,探讨其与冠状动脉支架内再狭窄(ISR)的关系。方法选取我院心内科经皮冠状动脉支架植入术后12个月内复查冠状动脉造影病人210例,根据造影结果分为支架内无再狭窄组(186例)和ISR组(24例)。检测血清miR-223和CXCR4表达水平,分析血清miR-223和CXCR4表达水平与ISR发生的关系。结果ISR组高血压占比、高血糖占比、总胆固醇和低密度脂蛋白胆固醇高于无再狭窄组(P<0.05)。ISR组血清miR-223和CXCR4水平高于无再狭窄组(P<0.05)。血清miR-223和CXCR4表达水平与ISR病人病变类型和长度有关(P<0.05)。Pearson相关性分析显示:血清miR-223、CXCR4水平与ISR病人血清总胆固醇、低密度脂蛋白胆固醇水平呈正相关(P<0.05)。Logistic分析结果表明,血清miR-223、CXCR4、总胆固醇和低密度脂蛋白胆固醇水平及高血压、高血糖为ISR发生的危险因素。结论ISR病人血清miR-223和CXCR4高表达,是冠状动脉ISR发生的危险因素。

生脉散联合阿托伐他汀对老年冠心病患者TLR4/MyD88/NF-κB信号通路的影响

目的:研究生脉散联合阿托伐他汀对老年冠心病(CHD)患者TLR4/MyD88/NF-κB信号通路的影响。方法:收集112例老年CHD患者作为研究对象,采用随机数字表法将患者分为观察组和对照组,每组56例。两组均接受常规治疗,对照组加用阿托伐他汀,观察组加用阿托伐他汀与生脉散。比较两组临床疗效、心功能指标[左室射血分数(LVEF)、左室舒张末内径(LVDD)及6 min步行距离(6MWD)],以及外周血TLR-4、MyD88、NF-κB mRNA和蛋白表达水平。结果:观察组总有效率高于对照组。治疗后,观察组TLR4、MyD88及NF-κB mRNA和蛋白表达低于对照组,差异有统计学意义(均P<0.05)。治疗后,观察组LVEF、6MWD水平高于对照组,差异有统计学意义(均P<0.05)。结论:生脉散联合阿托伐他汀治疗老年CHD疗效较好,可能与其调控TLR4/MyD88/NF-κB信号通路相关。

壮族冠心病患者的TLR4表达水平及其rs4986790A/G、rs4986791C/T位点的多态性分析

目的探讨广西壮族冠心病(CHD)患者的TLR4蛋白表达水平及其rs4986790A/G、rs4986791C/T位点的多态性。方法选取2016年1月至2018年1月在我院行冠状动脉造影(CAG)检查被确诊为CHD的壮族患者245例为CHD组,选取同期在我院住院治疗的245例非CHD患者为对照组。采用酶联免疫吸附法(ELISA)检测两组患者的血清TLR4蛋白表达水平;采用SNaPshot测序技术检测两组患者外周血TLR4基因rs4986790A/G、rs4986791C/T位点的基因型及等位基因。分析TLR4蛋白表达水平及其rs4986790A/G、rs4986791C/T位点的多态性与壮族人群CHD发病的相关性。结果CHD组患者的外周血TLR4蛋白表达水平高于对照组,差异有统计学意义(P<0.05)。CHD组患者的TLR4基因rs4986790A/G、rs4986791C/T位点的等位基因及基因型频率分布与对照组患者比较,差异无统计学意义(P>0.05)。结论外周血TLR4蛋白表达水平与广西壮族人群CHD的发生存在关联,但TLR4基因rs4986790A/G、rs4986791C/T位点的多态性可能与壮族人群CHD冠心病发病无明显的相关性。

冠心病合并2型糖尿病患者糖基化终产物与单核细胞TLR4的关系

目的探讨分析冠心病(CHD)合并糖尿病(DM)患者糖基化终产物(AGEs)与单核细胞toll样受体4(TLR4)的关系。方法 CHD+DM患者58例(CHD+DM组),CHD患者60例(CHD组),健康对照组50例。采用酶联免疫吸附测定(ELISA)法检测各组的AGEs,用流式细胞仪检测外周血单核细胞TLR4m RNA相对表达量及TLR4阳性单核细胞比率。结果与健康对照组比较,CHD+DM组及CHD组AGEs及外周血单核细胞TLR4m RNA相对表达量、TLR4阳性单核细胞比率明显增高(P<0.05);CHD+DM组与CHD组比较,CHD+DM组AGEs及外周血单核细胞TLR4m RNA相对表达量、TLR4阳性单核细胞比率增高(P<0.05);相关性分析显示升高的AGEs与升高的TLR4m RNA相对表达量及TLR4阳性单核细胞比率呈正相关。结论 CHD+DM患者AGEs及TLR4表达高于CHD患者;CHD患者AGEs及TLR4表达高于健康对照组。AGEs与TLR4的表达呈正相关。

湖南汉族人群中Toll样受体2和4基因遗传多态性与冠心病的关系

目的:探讨湖南地区汉族人群Toll样受体(Toll-like receptor 2,TLR2)及TLR4基因多态性与冠状动脉粥样硬化性心脏病(冠心病)的关系。方法:运用Sanger测序技术检测180例正常对照组及167例无血缘关系湖南汉族人群冠心病组中TLR2 SNP 2258A>G及TLR4 SNP896A>G和SNP1196C>T的多态性。结果:TLR2 SNP 2258A>G及TLR4SNP896A>G基因型频率和等位基因频率在冠心病组和正常对照组之间的分布差异无统计学意义(P>0.05),TLR4SNP1196C>T在两者之间差异有统计学意义(P<0.05)。结论:湖南地区汉族人群TLR4基因多态性位点SNP1196C>T与冠心病有关。

血清CTRP3、CXCR4水平与冠心病行PCI后患者支架内再狭窄的关系

目的探讨血清C1q/TNF相关蛋白3(CTRP3)、CXC族趋化因子受体4(CXCR4)水平与冠状动脉(冠脉)粥样硬化性心脏病(冠心病)行经皮冠脉介入术(PCI)后患者支架内再狭窄(ISR)的关系。方法选择2017年6月至2019年12月于郑州大学第二附属医院心血管内科收治的157例冠心病患者(冠心病组)和150例体检者(对照组),检测血清CTRP3、CXCR4水平,PCI后随访12个月,根据是否发生ISR将患者分为ISR组(31例)和NISR组(126例)。分析影响冠心病患者PCI后发生ISR的危险因素以及CTRP3、CXCR4预测冠心病患者PCI后发生ISR的价值。结果冠心病组、ISR组血清CTRP3、CXCR4水平分别低于对照组、NISR组(P<0.05)。高D-二聚体、低CTRP3、CXCR4水平是冠心病患者PCI后发生ISR的危险因素(P<0.01),校正D-Dimer影响后,CXCR4(OR=0.512,95%CI:0.412~0.639,P≤0.001)、CTRP3(OR=0.569,95%CI:0.497~0.712,P≤0.001)仍与冠心病患者PCI后ISR的发生有关。CTRP3、CXCR4、联合CTRP3和CXCR4预测冠心病患者PCI后发生ISR的曲线下面积(AUC)分别为0.890、0.788、0.939。结论冠心病患者血清CXCR4、CTRP3水平降低,低CXCR4,CTRP3与冠心病患者PCI后ISR的发生有关,可作为冠心病患者预后评估的参考指标。