Albumin-bound kynurenic acid is an appropriate endogenous biomarker for assessment of the renal tubular OATs-MRP4 channel

Renal tubular secretion mediated by organic anion transporters(OATs)and the multidrug resistanceassociated protein 4(MRP4)is an important means of drug and toxin excretion.Unfortunately,there are no biomarkers to evaluate their function.The aim of this study was to identify and characterize an endogenous biomarker of the renal tubular OATs-MRP4 channel.Twenty-six uremic toxins were selected as candidate compounds,of which kynurenic acid was identified as a potential biomarker by assessing the protein-binding ratio and the uptake in OAT1-,OAT3-,and MRP4-overexpressing cell lines.OAT1/3 and MRP4 mediated the transcellular vectorial transport of kynurenic acid in vitro.Serum kynurenic acid concentration was dramatically increased in rats treated with a rat OAT1/3(rOAT1/3)inhibitor and in rOAT1/3 double knockout(rOAT1/3^(-/-))rats,and the renal concentrations were markedly elevated by the rat MRP4(rMRP4)inhibitor.Kynurenic acid was not filtered at the glomerulus(99%of albumin binding),and was specifically secreted in renal tubules through the OAT1/3-MRP4 channel with an appropriate affinity(Km)(496.7 mM and 382.2 mM for OAT1 and OAT3,respectively)and renal clearance half-life(t1/2)in vivo(3.7±0.7 h).There is a strong correlation in area under the plasma drug concentration-time curve(AUC0et)between cefmetazole and kynurenic acid,but not with creatinine,after inhibition of rOATs.In addition,the phase of increased kynurenic acid level is earlier than that of creatinine in acute kidney injury process.These results suggest that albumin-bound kynurenic acid is an appropriate endogenous biomarker for adjusting the dosage of drugs secreted by this channel or predicting kidney injury.

ELABELA protects against diabetic kidney disease by activating high glucose-inhibited renal tubular autophagy

ELABELA(ELA),an endogenous ligand of the apelin receptor(also known as apelin peptide jejunum[APJ]),has been shown to decrease in the plasma of patients with diabetic kidney disease(DKD).In the current study,we explored the potential function as well as the underlying mechanisms of ELA in DKD.We first found that the ELA levels were decreased in the kidneys of DKD mice.Then,we found that ELA administration mitigated renal damage and downregulated the expression of fibronectin,collagenⅣ,and transforming growth factor-β1 in the db/db mice and the high glucose cultured HK-2 cells.Furthermore,the autophagy markers,Beclin-1 and LC3-Ⅱ/LC3-Ⅰratio,were significantly impaired in DKD,but the ELA treatment reversed these alterations.Mechanistically,the inhibitory effects of ELA on the secretion of fibrosis-associated proteins in high glucose conditions were blocked by pretreatment with 3-methyladenine(an autophagy inhibitor).In summary,these in vivo and in vitro results demonstrate that ELA effectively protects against DKD by activating high glucose-inhibited renal tubular autophagy,potentially serving as a novel therapeutic candidate for DKD.

Decreased TRPM7 alleviates high glucose-induced renal tubular epithelial cell injury by inhibiting the HMGB1/TLR4 signaling pathway

Objective:To explore the regulatory mechanism of transient receptor potential melastatin-7(TRPM7)in high glucose-induced renal tubular epithelial cell injury.Methods:The expression of TRPM7 in the serum of diabetic nephropathy patients and high glucose-induced HK-2 cells was detected by RT-qPCR.Then,the TRPM7 interference vector was constructed,and the downstream high mobility group box 1(HMGB1)/Toll-like receptor 4(TLR4)signaling pathway proteins were detected.Next,in addition to interference with TRPM7 expression,overexpression of HMGB1 in high glucose-induced HK-2 cells was performed.Cell activity,apoptosis,oxidative stress levels,and inflammation levels were determined by CCK8,TUNEL,Western blotting,immunofluorescence and related kits.Results:TRPM7 expression was upregulated in the serum of diabetic nephropathy patients and high glucose-induced HK-2 cells.Interference with TRPM7 reduced cell damage,epithelial-mesenchymal transition,oxidative stress,and inflammatory response in high glucose-induced HK-2 cells via inhibiting the HMGB1/TLR4 signaling pathway.However,the effects induced by TRPM7 silencing were abrogated by HMGB1 overexpression.Conclusions:Decreased TRPM7 alleviates high glucose-induced renal tubular epithelial cell injury by inhibiting the HMGB1/TLR4 signaling pathway.Further animal experiments and clinical trials are warranted to verify its effect.

Incomplete distal renal tubular acidosis uncovered during pregnancy:A case report

BACKGROUND Renal tubular acidosis(RTA)is a renal cause of non-anion-gap metabolic acidosis characterized by low urinary ammonia excretion.This condition has a low prevalence,and various congenital and acquired etiologies.To date,only a few cases of idiopathic RTA uncovered during pregnancy have been reported.CASE SUMMARY A previously healthy 32-year-old Korean woman at 30 wk of gestation was admitted to Pusan National University Hospital with preterm labor.At admission,the patient presented with hypokalemia,non-anion-gap metabolic acidosis,and nephrocalcinosis.Distal RTA was diagnosed based on laboratory blood and urine findings and imaging examinations.Various tests,including next-generation gene sequencing panels for nephropathy,were performed to determine the etiology of the disease,which indicated that it was idiopathic.The patient received sodium bicarbonate and potassium chloride supplementation.After 3 wk,she delivered a baby who was subsequently diagnosed with corpus callosum agenesis and colpocephaly.During regular follow-ups for 6 mo postpartum,her hypokalemia and metabolic acidosis were gradually resolved,and medications eventually discontinued.CONCLUSION Herein we describe a case of idiopathic distal RTA discovered during pregnancy.Hypokalemia and metabolic acidosis resolved spontaneously after delivery.

Renal calcification in children with renal tubular acidosis:What a paediatrician should know

Renal tubular acidosis(RTA)can lead to renal calcification in children,which can cause various complications and impair renal function.This review provides pediatricians with a comprehensive understanding of the relationship between RTA and renal calcification,highlighting essential aspects for clinical manage-ment.The article analyzed relevant studies to explore the prevalence,risk factors,underlying mechanisms,and clinical implications of renal calcification in children with RTA.Results show that distal RTA(type 1)is particularly associated with nephrocalcinosis,which presents a higher risk of renal calcification.However,there are limitations to the existing literature,including a small number of studies,heterogeneity in methodologies,and potential publication bias.Longitudinal data and control groups are also lacking,which limits our understanding of longterm outcomes and optimal management strategies for children with RTA and renal calcification.Pediatricians play a crucial role in the early diagnosis and management of RTA to mitigate the risk of renal calcification and associated complications.In addition,alkaline therapy remains a cornerstone in the treatment of RTA,aimed at correcting the acid-base imbalance and reducing the formation of kidney stones.Therefore,early diagnosis and appropriate therapeutic interventions are paramount in preventing and managing renal calcification to preserve renal function and improve long-term outcomes for affected children.Further research with larger sample sizes and rigorous methodologies is needed to optimize the clinical approach to renal calcification in the context of RTA in the pediatric population.

Role of Connective Tissue Growth Factor in Extracellular Matrix Degradation in Renal Tubular Epithelial Cells

In order to investigate the effects of connective tissue growth factor （CTGF） antisense oligodeoxynucleotide （ODN） on plasminogen activator inhibitor-1 （PAI-1） expression in renal tubular cells induced by transforming growth factor β1 （TGF-β1） and to explore the role of CTGF in the degradation of renal extracellular matrix （ECM）, a human proximal tubular epithelial cell line （HKC） was cultured in vitro. Cationic lipid-mediated CTGF antisense ODN was transfected into HKC. After HKC were stimulated with TGF-β1 （5 μg/L）, the mRNA level of PAI-1 was detected by RT-PCR. Intracellular PAI-1 protein synthesis was assessed by flow cytometry. The secreted PAI-1 in the media was determined by Western blot. The results showed that TGF-β1 could induce tubular CTGF and PAI-1 mRNA expression. The PAI-1 mRNA expression induced by TGF-β1 was significantly inhibited by CTGF antisense ODN. CTGF antisense ODN also inhibited intracellular PAI-1 protein synthesis and lowered the levels of PAI-1 protein secreted into the media. It was concluded that CTGF might play a crucial role in the degradation of excessive ECM during tubulointerstitial fibrosis, and blocking the biological effect of CTGF may he a novel way in preventing renal fibrosis.

The Effect of Connective Tissue Growth Factor on Human Renal Tubular Epithelial Cell Transdifferentiation

To investigate the role of connective tissue growth factor (CTGF) in transdifferentiation of human renal tubular epithelial cell (HKC), in vitro cultured HKC cells were divided into 3 groups: negtive control, low dose CTGF-treated group (rh CTGF, 2.5 ng/ml) and high dose CTGF-treated (rhCTGF, 5.0 ng/ml). Then the expression of α-smooth muscle actin (α-SMA) were assessed by indirect immuno-fluorescence, and the percentage of α-SMA positive cells were assessed by flow cytometry. RT-PCR were also performed to examine the mRNA level of α-SMA. Upon the stimulation of different concentrations of rhCTGF, the expression of α-SMA were markedly stronger than that in negative controls. The percentages of α-SMA positive cells were significantly higher in the stimulated groups than that of negative controls (38.9 %, 65.5 % vs 2.4 %, P<0.01) .α-SMA mRNA levels were also up-regulated by the stimulation of rhCTGF (P<0.01). These results suggest that CTGF can promote the transdifferentiation of human renal tubular epithelial cells towards myofibroblast (Myo-F).

Experimental Study on Detached Renal Tubular Epithelial Cells in Urine of Nephropathia Epidemic Patients

To elucidate the pathogenesis of acute renal failure (ARF) with nephropathia epidemic (NE), provide experimental evidence for the new therapy to NE and observe the effects of Arg-Gly-Asp (RGD) peptides on adhesion of re-nal tubular epithelial cell (RTEC), urine specimens of patients were collected un-der sterile conditions. Detached RTECs were separated, cultured and identified.Hantan Virus antigen was determined by using indirect immunofluorescence method and effects of RGD on adhesion of RTECs was observed by subgroup counting as well as by flow cytometry. This study showed that: (1) sublethal RTECs existed in the urine of NE-ARF patients, which could be cultured in monolayer form; (2 ) there was NE antigen in RTECs; and (3) adhesion of RTECs could be inhibited by RGD.

Inhibition of Ubiquitin-specific Protease 4 Attenuates Epithelial-Mesenchymal Transition of Renal Tubular Epithelial Cells via Transforming Growth Factor Beta Receptor Type Ⅰ

Objective Ubiquitin-specific protease 4(USP4)facilitates the development of transforming growth factor-beta 1(TGF-β1)-induced epithelial-mesenchymal transition(EMT)in various cancer cells.Moreover,EMT of renal tubular epithelial cells(RTECs)is required for the progression of renal interstitial fibrosis.However,the role of USP4 in EMT of RTECs remains unknown.The present study aimed to explore the effect of USP4 on the EMT of RTECs as well as the involved mechanism.Methods In established unilateral ureteral obstruction(UUO)rats and NRK-52E cells,immunohistochemistry and Western blot assays were performed.Results USP4 expression was increased significantly with obstruction time.In NRK-52E cells stimulated by TGF-β1,USP4 expression was increased in a time-dependent manner.In addition,USP4 silencing with specific siRNA indicated that USP4 protein was suppressed effectively.Meanwhile,USP4 siRNA treatment restored E-cadherin and weakened alpha smooth muscle actin(α-SMA)expression,indicating that USP4 may promote EMT.After treatment with USP4 siRNA and TGF-β1 for 24 h,the expression of TGF-β1 receptor type I(TβRI)was decreased.Conclusion USP4 promotes the EMT of RTECs through upregulating TβRI,thereby facilitating renal interstitial fibrosis.These findings may provide a potential target of USP4 in the treatment of renal fibrosis.

Gender Difference of Cadmium- induced Renal Tubular Dysfunction for Inhabitants in Toyama,Japan

Objective The aim of the present study mas to compare the gender difference for cadmium-induced renal tubular dysfunction between the male and female inhabitants. Methods Urinary β2-microglobulin was measured in 299 male (94%) and 342 female (92%) inhabitants aged 54-72 years,and the development of renal tubular dysfunction for 11 years was studied in the 62 married couples from them. Results A significantly higher cumulative incidence was found in both men and women in cadmium-polluted area,showing 68. 4% in men and 64. 8% in women compared to 15. 3% in men and 5. 9% in women in the reference areas. Relative risk of renal tubular dysfunction in females (11. 0) was higher than males (4. 5). The ratios of urinary β2-microglobulin and glucose were higher in women than those in men in both the cadmium-polluted areas and the reference areas. Conclusion Although almost identical incidences were detected between men and women, the changes in excretion of β2-microglobulin and glucose was greater in women than those in men. These findings suggest that renal tubular dysfunction might be more progressive in women than that in men.

Comparative proteomic analysis of renal tubular epithelial cell injury caused by oxalic acid and calcium oxalate monohydrate

Objective To analyze and identify the differentially expressed proteins in human renal tubular epithelial ceils ( HK-2) after injury caused by oxalic acid and calcium oxalate monohydrate ( COM ) crystal,and to explore the potential role of renal tubular cell injury in kidney stone formation. Methods Normal HK-2 cells

Effect and mechanism of adrenomedullin on apoptosis of renal tubular epithelial cell in rats induced by renal ischemia reperfusion injury

Objective To investigate the effect and mechanism of adrenomedullin ( AM ) on apoptosis of renal tubular epithelial cell in rats induced by renal ischemia reperfusion injury. Methods Thirty-two Wistar rats were randomly divided into 4 groups: control group,IRI group, empty plasmid group and AM group. One week after re-

Chronic hepatitis B serum promotes apoptotic damage in human renal tubular cells

瞄准：在肾的管状的上皮细胞在试管内的 apoptosis 上与长期的肝炎 B ( CHB )调查病人的浆液的效果并且在肝炎 B 的致病学习肝炎 B ( HBV )和转变生长因素贝它( 1 )(TGF贝它( 1 ))的角色病毒联系了肾小球性肾炎( HBV-GN )。方法：浆液 TGF 贝它(1 ) 的层次被特定的酶测量连接免疫吸着剂试金(ELISA ) 和 HBV DNA 被聚合酶链反应(PCR ) 作为控制与 CHB，和 20 个健康的人在 44 个病人测试。正常的人的肾近似管状的房间(HK-2 ) 和重量的单位是有教养的一 of 健康的人，有 HBV-DNA negative (20 个案例) 的 CHB 病人和 HBV-DNA 积极(24 个案例) 为多达 72 h。Apoptosis 和 HK-2 的船边交货表示被流动血细胞计数器检测。结果：HK-2 房间的 apoptosis 率和船边交货表达式在 HBV DNA 积极浆液组 19.01%+/-5.85% 和 17.58%+/-8.35% 是显著地更高的， HBV DNA 否定浆液在控制组 4.25%+/-0.65% 和 2.33%+/-1.09% 比那些组织 8.12%+/-2.80% 和 6.96%+/-2.76% ，分别地(P【0.01 ) 。在 HBV DNA 积极浆液组的 HK-2 的 apoptosis 率和船边交货表示比在 HBV DNA 否定浆液(P【0.01 ) 的那些显著地高。在 HBV DNA 积极浆液组的 HK-2 房间的 Apoptosis 率断然与 HBV-DNA 的水平被相关(r = 0.657 ) 。在 CHB 组的浆液 TGF 贝它(1 ) 的水平是 163.05+/-91.35 microg/L，象与在控制组(P【0.01 ) 的 81.40+/-40.75 microg/L 相比显著地更高。结论：有长期的肝炎 B 的病人的浆液由触发船边交货起来规定的一条小径在人的肾的管状的房间支持 apoptotic 损坏。HBV 和 TGF 贝它(1 ) 可以在肝炎 B 的机制起重要作用联系肾小球性肾炎。

Diabetes and renal tubular cell apoptosis

Apoptosis contributes to the development of diabetic nephropathy, but the mechanism by which high glucose induces apoptosis is not fully understood. Apoptosis of tubular epithelial cells is a major feature of diabetic kidney disease, and hyperglycemia triggers the generation of free radicals and oxidant stress in tubular cells. Hyperglycemia and high glucose in vitro also lead to apoptosis, a form of programmed cell death. High glucose similar to those seen with hyperglycemia in people with diabetes mellitus, lead to accelerated apoptosis, a form of programmed cell death characterized by cell shrinkage, chromatin condensation and DNA fragmentation, in variety of cell types, including renal proximal tubular epithelial cells.

Dihydroartemisinin attenuates ischemia/reperfusion-induced renal tubular senescence by activating autophagy

Acute kidney injury(AKI)is an important factor for the occurrence and development of CKD.The protective effect of dihydroartemisinin on AKI and and reported mechanism have not been reported.In this study,we used two animal models including ischemia-reperfusion and UUO,as well as a high-glucose-stimulated HK-2 cell model,to evaluate the protective effect of dihydroartemisinin on premature senescence of renal tubular epithelial cells in vitro and in vivo.We demonstrated that dihydroartemisinin improved renal aging and renal injury by activating autophagy.In addition,we found that co-treatment with chloroquine,an autophagy inhibitor,abolished the anti-renal aging effect of dihydroartemisinin in vitro.These findings suggested that activation of autophagy/elimination of senescent cell might be a useful strategy to prevent AKI/UUO induced renal tubular senescence and fibrosis.

Role of connective growth factor in plasminogen activator inhibitor-1 and fibronectin expression induced by transforming growth factor β1 in renal tubular cells

Background Connective tissue growth factor (CTGF) contributes greatly to renal tubulointerstitial fibrosis, which is the final event leading to end-stage renal failure. This study was designed to investigate the effects of CTGF antisense oligodeoxynucleotides (ODNs) on the expressions of plasminogen activator inhibitor-1 (PAI-1) and fibronectin in renal tubular cells induced by transforming growth factor β1 (TGF-β1) in addition to the role of CTGF in the accumulation and degradation of renal extracellular matrix (ECM).Methods A human proximal tubular epithelial cell line (HKC) was cultured in vitro. Cationic lipid-mediated CTGF antisense ODNs were transfected into HKC cells. After HKC cells were stimulated with TGF-β1 (5 μg/L), the mRNA levels of PAI-1 and fibronectin were measured by RT-PCR. Intracellular PAI-1 protein synthesis was assessed by flow cytometry. The secreted PAI-1 and fibronectin in the medium were determined by Western blot and ELISA, respectively.Results TGF-β1 was found to induce tubular CTGF, PAI-1, and fibronectin mRNA expression. PAI-1 and fibronectin mRNA expression induced by TGF-β1 was significantly inhibited by CTGF antisense ODNs. CTGF antisense ODNs also inhibited intracellular PAI-1 protein synthesis and lowered the levels of PAI-1 and fibronectin protein secreted into the medium.Conclusions CTGF may play a crucial role in the accumulation and degradation of excessive ECM during tubulointerstitial fibrosis, and transfecting CTGF antisense ODNs may be an effective way to prevent renal fibrosis.

Distal renal tubular acidosis:genetic causes and management

Background Distal renal tubular acidosis(dRTA)is a kidney tubulopathy that causes a state of normal anion gap metabolic acidosis due to impairment of urine acidification.This review aims to summarize the etiology,pathophysiology,clinical findings,diagnosis and therapeutic approach of dRTA,with emphasis on genetic causes of dRTA.Data sources Literature reviews and original research articles from databases,including PubMed and Google Scholar.Manual searching was performed to identify additional studies about dRTA.Results dRTA is characterized as the dysfunction of the distal urinary acidification,leading to metabolic acidosis.In pediatric patients,the most frequent etiology of dRTA is the genetic alteration of genes responsible for the codification of distal tubule channels,whereas,in adult patients,dRTA is more commonly secondary to autoimmune diseases,use of medications and uropathies.Patients with dRTA exhibit failure to thrive and important laboratory alterations,which are used to define the diagnosis.The oral alkali and potassium supplementation can correct the biochemical defects,improve clinical manifestations and avoid nephrolithiasis and nephrocalcinosis.Conclusions dRTA is a multifactorial disease leading to several clinical manifestations.Clinical and laboratory alterations can be corrected by alkali replacement therapy.

过表达线粒体蛋白磷酸酶2C对人肾小管上皮细胞转录组的影响

背景:课题组前期研究发现相较于野生型小鼠,线粒体蛋白磷酸酶2C(protein phosphatase 2Cm,PP2Cm)基因缺失小鼠明显发生肾功能衰竭的症状,因此推测PP2Cm可能在肾脏纤维化发展过程中发挥重要保护作用,然而其分子机制尚不明确。目的:探究PP2Cm基因对于人肾小管上皮细胞转录组的影响。方法:培养人肾小管上皮细胞,用质粒将PP2Cm基因转染进入人肾小管上皮细胞,荧光定量PCR实验和Western blot实验检测细胞中PP2Cm的表达,随后分别提取细胞RNA进行转录组测序,寻找转染组和对照组之间的差异性表达基因,利用生物信息学方法进一步对所得的差异基因进行GO分析和KEGG分析。结果与结论:通过测序分析,与未转染空白细胞相比,在转染PP2Cm基因的人肾小管上皮细胞中存在796个差异性表达基因,其中553个下调基因,243个上调基因,GO分析结果显示,上调表达的基因显著富集在细胞生物合成过程、蛋白质翻译、内在凋亡信号通路等;下调表达的基因显著富集在内皮细胞增殖、细胞黏附等信号通路;KEGG分析结果显示,显著上调表达的基因富集在氨基酸代谢、生物合成等代谢相关信号通路;下调表达的基因显著富集在泛酸和辅酶A的生物合成等信号通路。结果表明,PP2Cm过表达可以影响肾小管上皮细胞的一系列生物学过程相关的多条信号通路,可能在氨基酸代谢、生物合成等代谢相关信号通路中起重要作用。

肾小管及肾小球标志物在2型糖尿病患者不同肾损伤阶段诊断价值的研究

目的探讨肾小管及肾小球相关标志物在2型糖尿病(type 2 diabetes mellitus,T2DM)患者不同肾损伤阶段的诊断价值。方法选取于2018年4月1日至2019年10月31日入住首都医科大学附属北京同仁医院内分泌科的T2DM患者272例,完善临床生化指标及尿蛋白四项:尿微量白蛋白/肌酐(urinary albumin to creatinine ratio,ACR)、α1-微球蛋白/肌酐(urinary α1-microglobulin to creatinine ratio,UA1CR)、免疫球蛋白G/肌酐(urinary immunoglobulin G to creatinine ratio,UIGG)、转铁蛋白/肌酐(urinary transferrin to creatinine ratio,UTRF);进行眼底照相、核医学99mTc-EC检测肾有效血浆流量(effective renal plasma flow,ERPF)和99mTc-DTPA检测肾小球滤过率(glomerular filtration rate,GFR)。根据ACR和眼底检查结果分为4组:正常蛋白尿无糖尿病视网膜病变(diabetic retinopathy,DR)132例,即对照组(ACR≤30 mg/g);正常蛋白尿合并DR 32例,为糖尿病肾病(diabetic kidney disease,DKD)前期组;微量蛋白尿组78例(30<ACR≤300 mg/g)和大量蛋白尿组30例(ACR>300 mg/g)。比较四组间尿蛋白四项和ERPF、GFR的水平,通过受试者工作特征(receiver operating characteristic,ROC)曲线评价上述各指标在不同肾损伤阶段的诊断价值。结果尿蛋白四项和ERPF、GFR的水平在不同组间差异有统计学意义(P<0.05)。在尿蛋白正常组中,DR组中肾小管功能标志物UA1CR较对照组明显升高(P<0.01);肾小球功能标志物ACR、UTRF和GFR在两组间差异无统计学意义(P>0.05),DR组UIGG较对照组升高(P<0.01)。在微量蛋白尿组和大量蛋白尿组,尿蛋白四项随肾损伤程度增加而增加,而ERPF和GFR随肾损伤程度增加而降低。ROC曲线分析显示,在尿蛋白排出正常的T2DM患者中合并DR组中肾小管功能标志物UA1CR和ERPF的曲线下面积(area under the curve,AUC)分别为68.2%(P<0.01)和60.5%(P<0.05),而肾小球功能标志物ACR和GFR的AUC均小于60%,差异无统计学意义(P>0.05)。尿蛋白四项及GFR在微量和大量蛋白尿组的AUC均大于60%(P<0.05),ERPF在大量蛋白尿组AUC为67.2%(P<0.05)。结论T2DM极早期微血管改变即ACR正常仅有DR时,肾小管标志物UA1CR先于肾小球标志物ACR和GFR发生变化。肾损伤早期,肾小管标志物诊断效能优于肾小球;肾损伤后期,肾小球标志物诊断效能优于肾小管。提示DKD肾小管功能的改变可能早于肾小球。

MCP-1基因对脓毒症急性肾损伤发生的作用研究

目的研究单核细胞趋化蛋白-1(MCP-1)基因对脓毒症急性肾损伤(AKI)发生的作用。方法选取2022年9月-2023年9月新疆维吾尔自治区人民医院重症医学科收治的50例脓毒症AKI患者作为AKI组,纳入同期50例健康受试者作为对照组。分别采集全部受试者清晨空腹静脉血5 mL,采用ELISA法测定AKI患者及健康人群血清中MCP-1的表达情况;通过原代培养肾小管上皮细胞,利用CK14和CK18抗体进行细胞免疫荧光鉴定,过表达和干扰MCP-1基因,利用CCK8细胞增殖检测试剂盒和RT-qPCR检测肾小管上皮细胞增殖情况和肿瘤坏死因子α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素6(IL-6)、白细胞介素10(IL-10)、γ-干扰素(IFN-γ)炎性因子表达的变化。结果与对照组相比,AKI组患者外周血中和尿液中的MCP-1表达量显著升高(P均<0.05);通过细胞免疫荧光鉴定,选择上皮细胞标志物CK14和CK18,原代培养24 h,90%以上的细胞表达细胞标志物CK18,约84%的细胞表达CK14;与NC组相比,siRNA组在24、48 h细胞数量增加(P均<0.05);与MCP-1组相比,siRNA组在24、48、72 h的细胞数量增加(P均<0.05);NC组的IL-1β、IL-6、INF-γ因子的表达水平随时间推移逐渐升高,MCP-1组的IL-1β、IL-6、IL-10、INF-γ和TNF-α因子的表达水平随时间推移逐渐升高,siRNA组的IL-1β、IL-6、INF-γ因子表达水平随时间推移无明显升高趋势。结论MCP-1基因在脓毒症急性肾损伤的发病机制中可能发挥重要作用,该基因可能通过调节IL-1β等炎性细胞因子的表达来抑制肾小管上皮细胞的生长和增殖,从而参与脓毒症患者的AKI过程。