Mathematical Model and Experiment Validation of Fluid Torque by Shear Stress under Influence of Fluid Temperature in Hydro-viscous Clutch

The current design of hydro-viscous clutch（HVC） in tracked vehicle fan transmission mainly focuses on high-speed and high power. However, the fluid torque under the influence of fluid temperature can not be predicted accurately by conventional mathematical model or experimental research. In order to validate the fluid torque of HVC by taking the viscosity-temperature characteristic of fluid into account, the test rig is designed. The outlet oil temperature is measured and fitted with different rotation speed, oil film thickness, oil flow rate, and inlet oil temperature. Meanwhile, the film torque can be obtained. Based on Navier-Stokes equations and the continuity equation, the mathematical model of fluid torque is proposed in cylindrical coordinate. Iterative method is employed to solve the equations. The radial and tangential speed distribution, radial pressure distribution and theoretical flow rate are determined and analyzed. The models of equivalent radius and fluid torque of friction pairs are introduced. The experimental and theoretical results indicate that tangential speed distribution is mainly determined by the relative rotating speed between the friction plate and the separator disc. However, the radial speed distribution and pressure distribution are dominated by pressure difference at the lower rotating speed. The oil film fills the clearance and the film torque increases with increasing rotating speed. However, when the speed reaches a certain value, the centrifugal force will play an important role on the fluid distribution. The pressure is negative at the outer radius when inlet flow rate is less than theoretical flow, so the film starts to shrink which decreases the film torque sharply. The theoretical fluid torque has good agreement with the experimental data. This research proposes a new fluid torque mathematical model which may predict the film torque under the influence of temperature more accurately.

Possible effect of fluid shear stress on osteoclastogenesis

Bone remodeling is performed under the joint action of osteoblasts and osteoclasts. Since the effect of osteoclasts has been gradually recognized on bone and joint diseases, targeted researches toward osteoclasts have become a hot research field. This article reviews the relevant medical literature concerning the possible effects of the fluid shear stress (FSS) on the osteoclastogenesis chiefly from the aspects of RANKL-RANK-OPG system, the macrophage colony-stimulating factor (M-CSF), and calcitonin receptor (CTR). On the basis of the changes of the expression of osteoclastic activities, it is suggested that FSS is a potent, important regulator of bone metabolism.

The roles of focal adhesion and cytoskeleton systems in fluid shearstress-induced endothelial cell response

Focal adhesions are polyproteins linked to extracellular matrix and cytoskeleton,which play an important role in the process of transforming force signals into intracellular chemical signals and subsequently triggering related physiological or pathological reactions.The cytoskeleton is a network of protein fibers in the cytoplasm,which is composed of microfilaments,microtubules,intermediate filaments,and cross-linked proteins.It is a very important structure for cells to maintain their basic morphology.This review summarizes the process of fluid shear stress transduction mediated by focal adhesion and the key role of the cytoskeleton in this process,which focuses on the focal adhesion and cytoskeleton systems.The important proteins involved in signal transduction in focal adhesion are introduced emphatically.The relationship between focal adhesion and mechanical transduction pathways are discussed.In this review,we discuss the relationship between fluid shear stress and associated diseases such as atherosclerosis,as well as its role in clinical research and drug development.

Influence of Hartmann Number on Convective Flow of Maxwell Fluid between Two Hot Parallel Plates through Porous Medium Subject to Arbitrary Shear Stress at the Boundary

Natural convection flow of unsteady Maxwell fluid with the effects of constant magnetic force in the course of a porous media is investigated in this research work. Fluid motion between a channel of parallel plates is tempted by time dependent shear stress applied on one plate. The governing partial differential equations of a model under consideration are transformed into ordinary differential equations by Laplace transform method and then solved for temperature and velocity fields. The obtained results for temperature fields are expressed in terms of complementary error function. The influences of involved parameters likes Hartmann number, Grashf number, Prandlt number and porosity parameter, on temperature and velocity profiles are shown graphically. There is no such result regarding Maxwell fluid in the existing literature.

Fluid Shear Stress-Induced IL-8/CXCR Signaling Promotes Epithelial Mesenchymal Transition of Ovarian Cancer Cells

Objective Epithelial mesenchymal transition(EMT)plays a very important role in ovarian cancer metastasis,and IL-8 released from mechanosensitive cancer cells may contribute to the EMT process of solid carcinomas.In this study,we have explored IL-8 and its receptors signal transduction process of human ovarian cancer cells under conditions of FSS,and simultaneously detected the EMT process of ovarian cancer.Methods After the fluid shear stress was loaded,LightCyclerTM system and ELISA were employed to assay the IL-8 mRNA expression and protein production,respectively.Meanwhile,IL-8 reporter gene pEGFP1-IL8USCS was constructed for determining IL-8 gene transcriptional activation through gene transfer and flow cytometric analysis.RT-PCR,Northern blot and immunofluorescence were used to determine the expression of IL-8 receptor CXCR2 at mRNA and protein levels.IL-8 downstream signaling molecule NF-κB nuclear translocation was observed by immunocytofluoresent staining.Western blot was used to examine IκB phosphorylation and EMT-related protein.Results(1)The increase of IL-8 mRNA expression by shear stress was time-dependent.The expression increased when SKOV3 cells exposed to fluid shear stress for 1 h,reached the summits at 2 h,gradually decreased at 3 h and remained at a constant level at 4~12 h.Additionally,IL-8 expression was negatively associated with the intensity of shear stress.After SKOV3 cells were exposed to low fluid shear stress(1.5 dyne/cm2)for 1 h and 2 h,IL-8 mRNA expression increased near 68 and 52 times respectively as that of SKOV3 cells exposed to a high fluid shear stress of 5.0 dyne/cm^2.(2)The productions of IL-8 protein in SKOV3 cells subjected to shear stress were time-dependent.The secretion reached the summit when SKOV3 cells exposed to fluid shear stress for 5 h,then IL-8 secretion gradually decreased at 8 h of stimulation by shear stress.IL-8 secretion increased obviously when fluid shear stress(0.5,1.5,or 2.0 dyne/cm2)was exerted on SKOV3 cells for 1 h.Notablely,the secretion of IL-8 was the highest when SKOV3 cells subjected to fluid shear stress 1.5dyne/cm^2,which was near 6 or 7 times as that of SKOV3 cells subjected to high fluid shear stress(5.0 dyne/cm^2).(3)There was an increase in enhanced green fluorescent protein expression in pEGFPI-IL8USCS-transfected SKOV3 cells subjected to a fluid shear stress of 1.5 dyne/cm2 for 2 h,suggesting a flow shear stress induced IL-8 gene transcriptional activation;(4)CXCR2,which was constitutively present on the surface of SKOV3 cells,increased following exposure to fluid shear stress for 60 min.(5)Following the application of a shear stress of 1.5 dyne/cm^2,NF-κB p65 became detectable in the cell nuclei and Phosphorylated IκB in cell lysates increased significantly;(6)Compared with the control group,critical EMT-related proteins vimentin was upregulated,E-cadherin was downregulated after the application of the 1.5 dyne/cm2shear stress for 2 h,which suggested the EMT of ovarian cancer.Conclusions FSS triggered IL-8/CXCR2 signaling of SK-OV3 cells represents an early gene activation and the activation can be mediated through NF-κB.When the fluid shear stress-induced IL-8/CXCR signaling activated,the expression of EMT-related proteins changed.This observation suggested that fluid shear stress-induced IL-8 activation and the downstream signal pathways may have important contribution to the EMT process of ovarian cancer cells.

Shear Stress in MR Fluid with Small Shear Deformation in Bctlattic Structure

A theoretical model based on BCT lattice structure was developed. Resultant force in the BCT lattice structure was deduced, following the interaction force of two kinds of magnetic particles. According to empirical FroHlich-Kermelly law, the relationship between the magnetic induction and the magnetic field was discussed, and a predictive formula of shear stresses of the BCT lattice structure model was established for the case of small shear deformation. Compared with the experimental data for different particle volume fractions, the theoretical results of the shear stress indicate the effects of the saturation magnetization and the external magnetic field on the shear stress.

A New Modified Conductivity Model for Prediction of Shear Yield Stress of Electrorheological Fluids Based on Face-center Square Structure

A new modified conductivity model was established to predict the shear yield stress of electrorheological fluids (ERF). By using a cell equivalent method, the present model can deal with the face-center square structure of ERF. Combining the scheme of the classical conductivity model for the single-chain structure, a new formula for the prediction of the shear yield stress of ERF was set up. The influences of the separation distance of the particles, the volume fraction of the particles and the applied electric field on the shear yield stress were investigated.

A New Theoretical Model about Shear Stress in Magnetorheological Fluids with Small Shear Deformation

Based on the single-chain structure model of magnetorheological fluids, a formu la for the calculation of shear stresses was established. The interaction force of two magnetic particles in an infinite single-chain was deduced using a new theoretical model which is founded on Ampere' molecular curr ent hypothesis, dipole theory and Ampere' law. Furthermore, the resultant force on a particle was then deduced by taking into account of the action caused by al l the other particles in the single-chain. A predictive formula for shear stres ses was made corresponding to the case that MR fluids were sheared by a small an gle and the calculating results fit well on the order with the yield stresses of the commercial MR fluids.

Heat Transfer of Casson Fluid over a Vertical Plate with Arbitrary Shear Stress and Exponential Heating

The basic objective of this work is to study the heat transfer of Casson fluid of non-Newtonian nature.The fluid is considered over a vertical plate such that the plate exhibits arbitrary wall shear stress at the boundary.Heat transfers due to exponential plate heating and natural convection are due to buoyancy force.Magnetohydrodynamic(MHD)analysis in the occurrence of a uniform magnetic field is also considered.The medium over the plate is porous and hence Darcy’s law is applied.The governing equations are established for the velocity and temperature fields by the usual Boussinesq approximation.The problem is first written in dimensionless form using some useful non-dimensional quantities and then solved.The exact analysis is performed and hence solutions via integral transform are established.The analysis of various pertinent parameters on temperature distribution and velocity field are reported graphically.It is found that pours medium permeability parameter retards the fluid motion whereas,velocity decreases with increasing magnetic parameter.Velocity and temperature decrease with increasing Prandtl number whereas the Grashof number enhances the fluid motion.Further,it is concluded from this study that the results obtained here are more general and in a limiting sense several other solutions can be recovered.The Newtonian fluid results can be easily established by taking the Casson parameter infinitely large i.e.,whenβ→∞.

STEADY-SHEAR VISCOSITY AND TRANSIENT STRESS RESPONSE FOR ELASTO-THIXOTROPIC FLUIDS

A phenomenological model for dispersed systems which exhibit complex theological behaviour such as shear and time-dependent viscosity, yield stress, and elasticity is proposed. The model extends the Quemeda model to describe the viscosity function with a structural parameter λ which varies according to differ- ent kinetic orders of particle aggregation and segregation. The transient stress response is obtained by solving an instantaneous Maxwell model with an assumed shear modulus function G of the same form as the viscosity function η. Accuracy of the proposed model is verified experimentally with the results obtained for two oil (creosote)/water emulsions. The model that gives the best fit of experimental data appears to be the one with kinetic orders n = m = 2.

Wall shear stress in intracranial aneurysms and adjacent arteries

Hemodynamic parameters play an important role in aneurysm formation and growth. However, it is difficult to directly observe a rapidly growing de novo aneurysm in a patient. To investigate possible associations between hemodynamic parameters and the formation and growth of intracranial aneurysms, the present study constructed a computational model of a case with an internal carotid artery aneurysm and an anterior communicating artery aneurysm, based on the CT angiography findings of a patient. To simulate the formation of the anterior communicating artery aneurysm and the growth of the internal carotid artery aneurysm, we then constructed a model that virtually removed the anterior communicating artery aneurysm, and a further two models that also progressively decreased the size of the internal carotid artery aneurysm. Computational simulations of the fluid dynamics of the four models were performed under pulsatile flow conditions, and wall shear stress was compared among the different models. In the three aneurysm growth models, increasing size of the aneurysm was associated with an increased area of low wall shear stress, a significant decrease in wall shear stress at the dome of the aneurysm, and a significant change in the wall shear stress of the parent artery. The wall shear stress of the anterior communicating artery remained low, and was significantly lower than the wall shear stress at the bifurcation of the internal carotid artery or the bifurcation of the middle cerebral artery. After formation of the anterior communicating artery aneurysm, the wall shear stress at the dome of the internal carotid artery aneurysm increased significantly, and the wall shear stress in the upstream arteries also changed significantly. These findings indicate that low wall shear stress may be associated with the initiation and growth of aneurysms, and that aneurysm formation and growth may influence hemodynamic parameters in the local and adjacent arteries.

The Effects of Post-Stenotic Dilatations on the Flow of Couple Stress Fluid through Stenosed Arteries

The flow of incompressible couple stress fluid in a circular tube with stenosis and dilatations has been investigated. The stenosis was assumed to be axially symmetric and mild. The flow equations have been linearized and the expressions for the resistance to the flow, velocity, pressure drop, wall shear stress have been derived. The effects of various parameters on these flow variables have been investigated. It is found that the resistance to the flow and pressure drop increase with height of the stenosis and decrease with post stenotic dilatation. Pressure drop decreases with couple stress fluid parameter for both stenosis and post stenotic dilatation. Further, the wall shear stress increases with height of the stenosis and couple stress parameter but decreases with post stenotic dilatation and couple stress fluid parameter.

Caveolin-1介导流体剪切应力调控MC3T3-E1成骨细胞增殖和凋亡

背景:流体剪切应力在成骨细胞增殖和凋亡中起着重要作用。然而,Caveolin-1(Cav-1)是否参与成骨细胞中流体剪切应力诱导的增殖与凋亡过程尚不清楚。目的:探讨Cav-1在流体剪切应力调控成骨细胞增殖和凋亡中的作用。方法:选择生长状态良好的MC3T3-E1成骨细胞,加载不同时间(0,30,60,90 min)且强度为1.2 Pa的流体剪切应力,观察Cav-1蛋白的表达,筛选出时间为60 min的条件进行实验。将MC3T3-E1细胞分为:对照组、流体剪切应力组、流体剪切应力+pcDNA 3.1组(对照)、流体剪切应力+pcDNA Cav-1组(过表达质粒),分别采用流体剪切应力和过表达Cav-1等方式干预,通过q RT-PCR和Western blot检测MC3T3-E1细胞内增殖以及凋亡相关分子的表达量;通过CCK-8与Ed U实验检测MC3T3-E1细胞增殖活性;采用Hoechst 33258染色以及流式细胞术检测MC3T3-E1细胞凋亡情况。结果与结论:加载流体剪切应力后MC3T3-E1细胞中Cav-1的表达显著下调,且加载60 min表达水平最低。过表达Cav-1减弱了流体剪切应力促进MC3T3-E1细胞增殖及抑制细胞凋亡的效应。说明Cav-1在流体剪切应力调节成骨细胞增殖和凋亡过程中具有重要的作用,并可能为骨质疏松症提供潜在治疗策略。

基于计算流体动力学技术评估动脉瘤性蛛网膜下腔出血后迟发性脑缺血风险

目的基于计算流体动力学(computational fluid dynamics,CFD)技术,探究动脉瘤性蛛网膜下腔出血(anauryrmal subarachnoid hemorrhage,aSAH)后颅内血流动力学早期变化与迟发性脑缺血(delayed cerebral ischemia,DCI)发生的潜在关系。方法本研究前瞻性收集经CTA(CT Angiography)诊断并经DSA(Digital Subtracted Angiography)或手术证实为颅内动脉瘤破裂的患者。收集患者的临床和术前、术后的头颅CTA数据。基于患者的CTA原始图像构建个体化颅内血管模型,基于CFD技术模拟计算术前、术后的压力和壁剪切应力(wall shear stress,WSS)及PR(pressure ratio)、WSSR(WSS ratio)。通过随访影像学或临床体格检查判断是否发生DCI,并分为DCI组和无DCI组,分析比较两组间临床资料及血流动力学参数差异及其与DCI相关性。结果共入组51例患者,其中DCI组共15例患者(29.4%)DCI组较无DCI组,入院时收缩压(170.87±20.28 vs.1.58.19±28.06,P=0.037)、Pressure术后(19735.10±860.18vs.14606.06±11260.28,P=0.010)和PR(1.58±0.01vs.1.37±2.12,P=0.012)均较高。单因素分析显示血流动力学参数均无统计学差异,而多因素分析显示Pressure术前(OR=1,95%CI:0.999~1,P=0.017)、Prassure术后(QR=1.00.1,95%CI:1~1.001,P=0.007)、WSS术前(OR=1.096,95%CI:1.002~1.198,P=0.045)和WSS术后(OR=0.888,95%CI:0.806~0.979,P=0.017)对DCI的影响有统计学意义。ROC曲线显示,基于Pressure术前、Pressure术后、WSS术前及WSS术后模型对DCI的识别效能AUC为0.794,敏感度为73.33%,特异度为86.1%。结论aSAH后颅内血流动力学早期改变与DCI发生有关。

机械应力影响细胞焦亡的机制研究进展

细胞焦亡是一种由炎性小体传感器激活引起的细胞死亡,最终导致细胞膜的完整性丧失。细胞焦亡导致其胞内容物泄漏至胞外空间诱导趋化和炎症,因此在炎症和疾病的发生发展中具有重要作用。细胞长期处在复杂的机械环境中,机械刺激可来源于微环境,也可来自外力。适宜的机械应力有助于细胞正常生理活动的进行,但过度的机械应力会激活应激信号通路,如死亡通路,从而引起细胞组织损伤和炎症反应,因此本文就细胞焦亡的分子机制及机械应力影响细胞焦亡的具体机制进行文献回顾及综述,旨在为研究焦亡相关疾病提供依据。

滑膜细胞及软骨细胞释放HMGB1流体力学阈值的研究

目的探究流体流动剪切力(fluid flow shear stress,FFSS)下,滑膜细胞及软骨细胞释放高迁移率族蛋白1(high⁃mobility group box 1,HMGB1)时流体力学的阈值;阐明异常机械力刺激下,滑膜细胞及软骨细胞破坏的先后顺序,为了解颞下颌关节骨关节炎的发病机制及病理研究提供实验基础。方法获得医院动物实验伦理委员会的审批,于SD大鼠膝关节获取滑膜组织及软骨组织块,消化培养获得滑膜细胞及软骨细胞,取3~4代滑膜细胞及软骨细胞,通过流体剪切力学装置对软骨及滑膜细胞施加FFSS,根据不同大小的FFSS值分组,分别采用1、3、5、10 dyn/cm^(2)的FFSS刺激滑膜细胞1 h,采用4、8、12、16 dyn/cm^(2)的FFSS刺激软骨细胞1 h,静息培养(0 dyn/cm^(2))作为对照组,观察细胞的形态学变化、免疫组化检测HMGB1及白细胞介素⁃1β(interleu⁃kin⁃1β,IL⁃1β)的细胞表达分布,ELISA检测上清液中HMGB1与IL⁃1β的水平;Western blot检测细胞HMGB1与IL⁃1β蛋白表达水平。结果随着FFSS负载的增大,滑膜细胞及软骨细胞逐渐肿胀,破裂,且细胞数量减少。随着FFSS负载的增大,HMGB1与IL⁃1β的表达由胞核逐渐转移至胞浆。在滑膜细胞中,与对照组相比,1、3、5、10 dyn/cm^(2)干预组中的HMGB1和IL⁃1β在上清液及细胞内的表达水平明显升高(P<0.01)。在软骨细胞中,与对照组相比,4、12、16 dyn/cm^(2) FFSS干预组中HMGB1在上清液中的表达水平增加(P<0.05),细胞HMGB1蛋白表达水平明显增加(P<0.01);而在8 dyn/cm^(2) FFSS干预组中HMGB1在上清液中的表达水平明显增加(P<0.01),但细胞HMGB1蛋白表达减少(P<0.01);与对照组相比,4、8、12、16 dyn/cm^(2) FFSS干预组中IL⁃1β在上清液中表达水平逐渐增加(P<0.01);除4 dyn/cm^(2)组外,随着FFSS负载增大,细胞中IL⁃1β的蛋白表达水平逐渐升高。结论滑膜细胞及软骨细胞随FFSS负载增大出现溶胀破裂,释放HMGB1流体力学阈值分别接近于1 dyn/cm^(2)和8 dyn/cm^(2);即在受到机械力刺激时,滑膜细胞先于软骨细胞损伤。

不同运动状态下胰岛素对2型糖尿病大鼠骨髓间充质干细胞分化的影响

目的研究正常活动和剧烈运动下胰岛素治疗对2型糖尿病大鼠股骨远端松质骨中固体受力和流体流动特性、骨髓间充质干细胞(bone marrow stromal cells,BMSCs)分化的影响。方法基于Micro-CT扫描图像构建正常活动和剧烈运动下4周和8周胰岛素治疗实验的对照组、糖尿病组、治疗组和安慰剂组大鼠股骨远端松质骨和流体的有限元模型,并利用流固耦合数值模拟方法,分析各组模型的力学参数和细胞分化参数;并对力学参数、细胞分化参数与形态学参数进行相关性分析。结果正常活动和剧烈运动下,胰岛素治疗改善了2型糖尿病大鼠的固体和流体力学参数、BMSCs分化参数。对于4周实验,正常活动与剧烈运动下胰岛素治疗分别使2型糖尿病大鼠的骨分化面积从64.024%增加至69.372%,以及从73.225%增加至75.336%;对于8周实验,正常活动与剧烈运动下胰岛素治疗分别使2型糖尿病大鼠的骨分化面积从67.239%增加至72.910%,以及从76.147%增加至78.291%。形态学参数中的BV/TV、Tb.N、Tb.Th、Tb.Sp、SMI与骨、软骨分化面积均具有显著相关性(P<0.05)。结论剧烈运动下8周胰岛素治疗组松质骨表面的BMSCs更容易分化形成骨组织。研究结果对进一步理解正常活动和剧烈运动下胰岛素对骨的影响具有重要意义,并为临床上治疗2型糖尿病时胰岛素治疗周期和运动方式的选择提供理论指导。

流体剪应力对人脐静脉内皮细胞葡萄糖调节蛋白78和C/EBP同源蛋白表达的影响

目的探索流体剪应力对人脐静脉内皮细胞(HUVECs)葡萄糖调节蛋白78(GRP78)和C/EBP同源蛋白(CHOP)表达的影响。方法以HUVECs作为实验细胞,设计并构建流体动力学模拟实验系统,控制流体动力学模拟实验系统中灌流液的流速,以实现对实验细胞施加不同的流体剪应力。按实验细胞在实验系统中所承受的不同流体剪应力,将实验细胞分为低剪应力组(A组;0.4 Pa)、中剪应力组(B组;0.8 Pa)和高剪应力组(C组;1.2 Pa)。每组HUVECs包含3个细胞玻片,每个玻片经实验系统灌流液反复循环流经12 h。采用蛋白质印迹法对各组细胞中GRP78和CHOP蛋白水平进行检测,采用实时荧光定量逆转录聚合酶链反应技术测定各组细胞GRP78和CHOP蛋白及其信使RNA(mRNA)相对水平。应用GraphPad Prism 8.0软件对数据进行统计学分析。结果(1)A、B、C组HUVECs中GRP78蛋白相对表达水平分别为1.33±0.46、0.93±0.34、0.64±0.30;多组间比较差异有统计学意义(F=36.17,P<0.05)。A组GRP78蛋白相对表达水平高于B组、C组(均P<0.01),B组GRP78蛋白相对表达水平高于C组(P=0.0013)。3组HUVECs中CHOP蛋白相对表达水平分别为:A组1.29±0.38,B组0.90±0.34,C组0.59±0.29;多组间比较差异有统计学意义(F=41.27,P<0.05)。A组CHOP蛋白相对表达水平高于B组、C组(均P<0.01),B组CHOP蛋白相对表达水平高于C组(P=0.0004)。(2)A、B、C组HUVECs中GRP78 mRNA相对表达水平分别为18.3±3.4、11.3±1.8、5.4±2.2;多组间比较差异有统计学意义(F=189.20,P<0.05)。A组GRP78 mRNA相对表达水平高于B组、C组(均P<0.01),B组GRP78 mRNA相对表达水平高于C组(P<0.01)。3组HUVECs中CHOP mRNA相对表达水平分别为:A组20.4±3.8,B组14.2±2.1,C组7.8±1.3;多组间比较差异有统计学意义(F=171.80,P<0.05)。A组CHOP mRNA相对表达水平高于B组、C组(均P<0.01),B组CHOP mRNA相对表达水平高于C组(P<0.01)。结论低流体剪应力可能增加HUVECs中GRP78、CHOP的蛋白及其mRNA表达水平。

异常流体剪切力通过cGAS-STING信号通路参与髓核细胞的凋亡、炎症与自噬

目的探讨异常流体剪切力诱导髓核细胞(NPC)凋亡、炎症和自噬的作用及其可能的机制。方法对NPC加载24 dyn/cm^(2)流体剪切力,时间分别为0、60、90、120、150 min。使用细胞骨架染色研究流体剪切力对细胞骨架及细胞形态的影响;使用5-乙炔基-2'-脱氧尿苷染色验证NPC的增殖能力;用线粒体膜电位变化评估流体剪切力对线粒体的损伤效应;用赫斯特染色研究凋亡与流体剪切力的关系;用丹酰戊二胺染色研究自噬与流体剪切力的关系;探究细胞培养基酸碱度的变化,验证流体剪切力与炎症的关联;使用蛋白质印迹法研究cGAS-STING相关蛋白(cGAS、STING、TBK1)、炎症相关蛋白(肿瘤坏死因子-α、COX-2)、凋亡相关蛋白(Bcl-2、Bax)以及自噬蛋白(IL3-Ⅰ/Ⅱ、p62)的变化。结果作为一种细胞间的机械刺激,24 dyn/cm^(2)强度的流体剪切力可造成NPC形态由梭形转变为多边形,细胞骨架发生重组,细胞增殖能力下降,培养基酸性增强,线粒体JC-1多聚体减少、单体增加;同时,细胞内肿瘤坏死因子-α、COX-2、Bax蛋白表达量上调,LC3Ⅱ/LC3Ⅰ增加,p62、Bcl-2降解增加,出现凋亡、炎症、自噬以及线粒体损伤;加载24 dyn/cm^(2)的流体剪切力可以激活NPC内的cGAS-STING信号通路,且与时间呈正相关关系,120 min时强度最高。使用cGAS抑制剂RU.521可以减缓24 dyn/cm^(2)流体剪切力造成的NPC的凋亡、炎症、自噬以及线粒体损伤。结论流体剪切力诱导的NPC凋亡、炎症和自噬与cGAS-STING信号通路激活相关,抑制cGAS-STING信号通路的激活可以缓解异常流体剪切力造成的细胞损害。

Single-cell transcriptomic analysis reveals transcript enrichment in oxidative phosphorylation,fluid sheer stress,and inflammatory pathways in obesity-related glomerulopathy

Obesity-related glomerulopathy(ORG)is an independent risk factor for chronic kid-ney disease and even progression to end-stage renal disease.Efforts have been undertaken to elucidate the mechanisms underlying the development of ORG and substantial advances have been made in the treatment of ORG,but relatively little is known about cell-specific changes in gene expression.To define the transcriptomic landscape at single-cell resolution,we analyzed kidney samples from four patients with ORG and three obese control subjects without kidney disease using single-cell RNA sequencing.We report for the first time that immune cells,including T cells and B cells,are decreased in ORG patients.Further analysis indicated that SPP1 was significantly up-regulated in T cells and B cells.This gene is related to inflammation and cell proliferation.Analysis of differential gene expression in glomerular cells(endothelial cells,mesangial cells,and podocytes)showed that these cell types were mainly enriched in genes related to oxidative phosphorylation,cell adhesion,thermogenesis,and inflammatory pathways(PI3K-Akt signaling,MAPK signaling).Furthermore,we found that the podocytes of ORG patients were enriched in genes related to the fluid shear stress pathway.Moreover,an evaluation of cell-cell communications revealed that there were interactions between glomerular parietal epithelial cells and other cells in ORG patients,with major interactions between parietal epithelial cells and podocytes.Altogether,our identification of molecular events,cell types,and differentially expressed genes may facilitate the development of new preventive or therapeutic approaches for ORG.