Peroxisome proliferator-activated receptors for hypertension

Peroxisome proliferator-activated receptors(PPARs) are ligand-activated transcription factors belonging to the nuclear receptor superfamily, which is composed of four members encoded by distinct genes(α, β, γ, and δ). The genes undergo transactivation or transrepression under specific mechanisms that lead to the induction or repression of target gene expression. As is the case with other nuclear receptors, all four PPAR isoforms contain five or six structural regions in four functional domains; namely, A/B, C, D, and E/F. PPARs have many functions, particularly functions involving control of vascular tone, inflammation, and energy homeostasis, and are, therefore, important targets for hypertension, obesity, obesity-induced inflammation, and metabolic syndrome in general. Hence, PPARs also represent drug targets, and PPARα and PPARγ agonists are used clinically in the treatment of dyslipidemia and type 2 diabetes mellitus, respectively. Because of their pleiotropic effects, they have been identified as active in a number of diseases and are targets for the development of a broad range of therapies for a variety of diseases. It is likely that the range of PPARγ agonist therapeutic actions will result in novel approaches to lifestyle and other diseases. The combination of PPARs with reagents or with other cardiovascular drugs, such as diuretics and angiotensin Ⅱ receptor blockers, should be studied.This article provides a review of PPAR isoform characteristics, a discussion of progress in our understanding of the biological actions of PPARs, and a summary of PPAR agonist development for patient management. We also include a summary of the experimental and clinical evidence obtained from animal studies and clinical trials conducted to evaluate the usefulness and effectiveness of PPAR agonists in the treatment of lifestyle-related diseases.

New insights into sodium transport regulation in the distal nephron:Role of G-protein coupled receptors

The renal handling of Na^+ balance is a major determinant of the blood pressure(BP) level. The inability of the kidney to excrete the daily load of Na+ represents the primary cause of chronic hypertension. Among the different segments that constitute the nephron, those present in the distal part(i.e., the cortical thick ascending limb, the distal convoluted tubule, the connecting and collecting tubules) play a central role in the fine-tuning of renal Na^+ excretion and are the target of many different regulatory processes that modulate Na^+ retention more or less efficiently. G-protein coupled receptors(GPCRs) are crucially involved in this regulation and could represent efficient pharmacological targets to control BP levels. In this review, we describe both classical and novel GPCR-dependent regulatory systems that have been shown to modulate renal Na^+ absorption in the distal nephron. In addition to the multiplicity of the GPCR that regulate Na^+ excretion, this review also highlights the complexity of these different pathways, and the connections between them.

Insulin receptors in the kidneys in health and disease

Insulin is an important hormone that affects various metabolic processes,including kidney function.Impairment in insulin's action leads to insulin resistance in the target tissue.Besides defects in post-receptor insulin signaling,impairment at the receptor level could significantly affect insulin sensitivity of the target tissue.The kidney is a known target of insulin;however,whether the kidney develops "insulin resistance" is debatable.Regulation of the insulin receptor(IR) expression and its function is very well studied in major metabolic tissues like liver,skeletal muscles,and adipose tissue.The physiological relevance of IRs in the kidney has recently begun to be clarified.The credit goes to studies that showed a wide distribution of IR throughout the nephron segments and their reduced expression in the insulin resistance state.Moreover,altered renal and systemic metabolism observed in mice with targeted deletion of the IR from various epithelial cells of the kidney has strengthened this proposition.In this review,we recapitulate the crucial findings from literature that have expanded our knowledge regarding the significance of the renal IR in normal-and insulin-resistance states.

The interplay between non-esterified fatty acids and bovine peroxisome proliferator-activated receptors: results of an in vitro hybrid approach

Background: In dairy cows circulating non-esterified fatty acids(NEFA) increase early post-partum while liver and other tissues undergo adaptation to greater lipid metabolism, mainly regulated by peroxisome proliferator-activated receptors(PPAR). PPAR are activated by fatty acids(FA), but it remains to be demonstrated that circulating NEFA or dietary FA activate bovine PPAR. We hypothesized that circulating NEFA and dietary FA activate PPAR in dairy cows.Methods: The dose-response activation of PPAR by NEFA or dietary FA was assessed using HP300 e digital dispenser and luciferase reporter in several bovine cell types. Cells were treated with blood plasma isolated from Jersey cows before and after parturition, NEFA isolated from the blood plasma, FA released from lipoproteins using milk lipoprotein lipase(LPL), and palmitic acid(C16:0). Effect on each PPAR isotype was assessed using specific synthetic inhibitors.Results: NEFA isolated from blood serum activate PPAR linearly up to ~ 4-fold at 400 μmol/L in MAC-T cells but had cytotoxic effect. Addition of albumin to the culture media decreases cytotoxic effects of NEFA but also PPAR activation by ~ 2-fold. Treating cells with serum from peripartum cows reveals that much of the PPAR activation can be explained by the amount of NEFA in the serum(R~2 = 0.91) and that the response to serum NEFA follows a quadratic tendency, with peak activation around 1.4 mmol/L. Analysis of PPAR activation by serum in MAC-T, BFH-12 and BPAEC cells revealed that most of the activation is explained by the activity of PPARδ and PPARγ, but not PPARα. Palmitic acid activated PPAR when added in culture media or blood serum but the activation was limited to PPARδ and PPARα and the response was nil in serum from post-partum cows. The addition of LPL to the serum increased > 1.5-fold PPAR activation.Conclusion: Our results support dose-dependent activation of PPAR by circulating NEFA in bovine, specifically δand γ isotypes. Data also support the possibility of increasing PPAR activation by dietary FA;however, this nutrigenomics approach maybe only effective in pre-partum but not post-partum cows.

Comparative studies of D_2 receptors and brain perfusion inhemi-parkinsonism rats

The relationship between dopamine D2 receptors and brain perfusion in hemi-parkinsonism rats was studied. Hemi-parkinsonism rats were made by stereotaxic 6-hydroxy dopamine (6-OH-DA) lesions in substantia nigra(SN) and ventral tegmental area(VTA), apomorphine (Apo) which could induced the successful model rat rotates toward the intact side was used to select the rats, 125I-IBZM ex-vivo autoradiography analysis and 99mTc-HM-PAO regional cerebral biodistribution were used to evaluate D2 receptors and cerebral blood flow. The HPLC-ECD were used to measure striatum DA and its metabolites content. The lesioned side striatum DA and its metabolites HVA DOPAC reduced significantly than that of the intact side and seudo-operated group, striatum/cerebellum 125I-IBZM uptake ratio was 8.04+-0.71 in lesioned side of hemi-parkinsonism rats, significantly increased compared with the intact side and the seudo-operated group(p <0.05), 30.11+-4.53% enhancement as compared to the intact side, and also show good correlation with 30 min Apo induced rotation numbers (r=0.98), the regional cerebral blood flow study didn’t show significant difference between bilateral brain cortex area(p >0.05). These results indicated that in the 6- OH-DA lesioned side DA content decreased significantly and an up-regulation of striatum D2 receptor binding sites was induced in hemi-parkinsonism rats, which showed good correlation with rotation behavior induced by Apo. Comparing with cerebral blood flow, D2 receptor reflected by IBZM seems to be more specific and earlier to detect the cerebral functional impairment in experimental bend-parkinsonism.

Effect of endothelin and endothelin A receptors on regional cerebral blood flow after traumatic brain injury in rabbits

Objective: To investigate the effect of endothelin and endothelin A receptors (ETAR) on regional cerebral blood flow after traumatic brain injury (TBI). Methods: The changes of endothelin 1 (ET 1) content with radioimmunoassay, mRNA expression and the location of ETAR with in situ hybridization, and the function and effect of antagonist BQ123 on regional cerebral blood flow (rCBF) through intracisternal application were dynamically observed on 130 adult rabbits after TBI. Results: ET 1 increased significantly in regional brain tissues, and the expression of ETAR mRNA increased apparently and predominantly distributed in the cerebromicrovascular endothelium after trauma. The rCBF declined significantly, but by using selective ETAR antagonist BQ123 to treat the rabbits, the decrease of rCBF could be apparently prevented. Conclusions: It demonstrates that ET 1 may primarily contribute to the rCBF decrease after TBI, while providing that the role of ET 1 is mediated through ETAR.

Glucagon-like peptide-1 receptor agonists as a possible intervention to delay the onset of type 1 diabetes:A new horizon

Type 1 diabetes(T1D)is a chronic autoimmune condition that destroys insulinproducing beta cells in the pancreas,leading to insulin deficiency and hyperglycemia.The management of T1D primarily focuses on exogenous insulin replacement to control blood glucose levels.However,this approach does not address the underlying autoimmune process or prevent the progressive loss of beta cells.Recent research has explored the potential of glucagon-like peptide-1 receptor agonists(GLP-1RAs)as a novel intervention to modify the disease course and delay the onset of T1D.GLP-1RAs are medications initially developed for treating type 2 diabetes.They exert their effects by enhancing glucose-dependent insulin secretion,suppressing glucagon secretion,and slowing gastric emptying.Emerging evidence suggests that GLP-1RAs may also benefit the treatment of newly diagnosed patients with T1D.This article aims to highlight the potential of GLP-1RAs as an intervention to delay the onset of T1D,possibly through their potential immunomodulatory and anti-inflammatory effects and preservation of beta-cells.This article aims to explore the potential of shifting the paradigm of T1D management from reactive insulin replacement to proactive disease modification,which should open new avenues for preventing and treating T1D,improving the quality of life and long-term outcomes for individuals at risk of T1D.

Expression of Toll-like Receptor 9 in Peripheral Blood Mononuclear Cells from Patients with Different Hepatitis B and C Viral Loads

The aim of the present study was to investigate the expression of toll-like receptors （TLR） 9 in peripheral blood mononuclear cells （PBMC） of patients with chronic hepatitis B and C with different virus copies. The study group included 90 patients （60 with chronic hepatitis B, and 30 with chronic hepatitis C）, and 20 healthy people served as control group. The protein and mRNA levels of TLR9 were detected by using flow cytometry and real-time PCR. The serum viral copies of HBV and HCV were measured in all patients, and the correlation between HBV-DNA copies or HCV-RNA copies and the TLR9 expression was analyzed. Our results demonstrated that HBV or HCV infection led to a decreased expression of TLR9 mRNA and protein compared to the control group （P〈0.05）. The TLR9 protein and mRNA levels were negatively correlated with serum viral copies of HBV and HCV （r=-0.632, r=-0.909, P〈0.01）. It was concluded that TLR9 mRNA and protein are down-regulated in PBMC of HBV-infected or HCV-infected patients, and they are negatively correlated with serum viral copies and play an important role in detecting viral replication of HBV and HCV.

Expression of c-kit receptor in peripheral blood mononuclear cells in patients with systemic lupus erythematosus

Objective： To determine the expression of c-kit receptor in peripheral blood mononuclear cells （PBMCs） in patients with systemic lupus erythematosus （SLE）, and analyze the relationship between the c-kit expression level of PBMCs and clinical parameters. Methods： Peripheral blood mononuclear cells in 47 patients with SLE and 21 healthy volunteers were collected. Expression of c-kit mRNA in PBMCs were determined with reverse transcription-polymerase chain reaction （RT-PCR）. The protein of c-kit receptor （CDllT） in PBMCs was measured by flow cytometry. Results： Expression of c-kit receptor protein and mRNA in patients with active or inactive SLE （ n = 47） were significantly higher than those in controls. The c-kit receptor of PBMCs in SLE patients were significantly higher than those in healthy controls （ n = 21 ）, the c-kit receptor of PBMCs in active patients （ n = 27） were significantly higher than those in inactive patients （ n = 20） and there was no significant difference was found between patients with inactive SLE and healthy controls（ P 〉 0.05）. The c-kit receptor of PBMCs in SLE have significant association with activity index. Conclusion： Production of c-kit receptor is aberrantly increased in PBMCs in patients with SLE. C-kit receptor might be more closely related to the clinical parameters in SLE patients, which might reflect the clinical status of SLE patients.

S3B-2 D1 Receptor-Mediated Endogenous tPA Upregulation Contributes to Acute Blood Brain Barrier Damage

Background:Disruption of the blood brain barrier(BBB)integrity at the early stage of ischemia is becoming a critical target to reduce hemorrhage transformation(HT)because of the potential to predict HT.However,the mechanism underlying early BBB damage is not very clear.It was reported that after acute ischemia,there was a significant increase of dopamine release in striatum where we have reported BBB damage as well as upregulation of HIF-1αafter 2 h ischemia.Objective:In current study,we aimed to investigate the role of dopamine signal pathway in BBB damage after acute ischemia using in vivo rat middle cerebral artery occlusion(MCAO)model.Results:Our data showed that there was an increase of endogenous tissue plasminogen activator(tPA)in BBB damage area and intra-striatum infusion of tPA inhibitor neuroserpin,significantly alleviated ischemia-induced BBB damage.In addition,intra-striatum infusion of D1 antagonist SCH23390 significantly decreased ischemia-induced upregulation of endogenous tPA,accompanied by decrease of BBB damage and occludin degradation.More important,inhibition of HIF-1 with inhibitor YC-1 significantly decreased acute ischemia-induced endogenous tPA upregulation and BBB damage.Conclusion:Taken together,our data demonstrate that acute ischemia disrupted BBB through activation of endogenous tPA via HIF-1 upreguationinduced dopamine increase,thus representing a new therapeutic target for protecting BBB,and may help alleviate HT following thrombolysis after ischemia stroke.

AT1 receptor downregulation:A mechanism for improving glucose homeostasis

There is a pathophysiological correlation between arterial hypertension and diabetes mellitus, established since the pre-diabetic state in the entity known as insulin resistance. It is known that high concentrations of angiotensin-Ⅱ enable chronic activation of the AT1 receptor, promoting sustained vasoconstriction and the consequent development of high blood pressure. Furthermore, the chronic activation of the AT1 receptor has been associated with the development of insulin resistance. From a molecular outlook, the AT1 receptor signaling pathway can activate the JNK kinase. Once activated, this kinase can block the insulin signaling pathway, favoring the resistance to this hormone. In accordance with the previously mentioned mechanisms, the negative regulation of the AT1receptor could have beneficial effects in treating metabolic syndrome and type 2diabetes mellitus. This review explains the clinical correlation of the metabolic response that diabetic patients present when receiving negatively regulatory drugs of the AT1 receptor.

益气养阴化痰祛瘀法对甲状腺机能亢进症激素抗体及氧化应激因子表达的影响

目的研究益气养阴化痰祛瘀法对甲状腺机能亢进症(Hyperthyroidism,简称甲亢)患者激素抗体和氧化应激因子表达的影响。方法选取2022年1月—2023年12月收治的76例甲亢患者,按照随机数字表法分组,对照组(38例)采用常规西药治疗,观察组(38例)在对照组基础上加用益气养阴化痰祛瘀方治疗;治疗2个月后对比两组临床疗效、中医证候积分、甲状腺功能(各项激素水平)、氧化应激因子、促甲状腺激素受体抗体(TRAb)、甲状腺过氧化物酶抗体(TPOAb)的表达。结果观察组治疗总有效率为94.74%(36/38),高于对照组(76.32%,29/38),差异有统计学意义(P<0.05)。治疗后,两组主症积分、次症积分和舌脉积分均降低,观察组低于对照组;两组促甲状腺激素(TSH)上升,游离三碘甲状腺原氨酸(FT3)、游离甲状腺素(FT4)下降(P<0.05),观察组改善高于对照组;两组超氧化物歧化酶(SOD)升高,观察组SOD水平高于对照组,差异有统计学意义(P<0.05)。结论益气养阴化痰祛瘀法用于甲亢患者的治疗取得确切治疗成果,可以有效改善甲状腺功能,通过调控甲状腺激素水平、TRAb、TPOAb以及氧化应激因子的表达以改善病情,缓解症状。

血清肾胺酶、尿调节素、可溶性尿激酶受体联合检测对糖尿病肾损伤早期诊断的价值分析

目的探究血清肾胺酶(RNLS)、尿调节素(UMOD)、可溶性尿激酶型纤溶酶原激活物受体(suPAR)联合检测对糖尿病肾损伤早期诊断的价值。方法选取2020年3月至2021年3月成都市第七人民医院收治的61例糖尿病肾损伤病人为糖尿病肾损伤组,按24 h尿白蛋白排泄率(UAER)分为早期糖尿病肾病组(29例)和临床期糖尿病肾病组(32例),选取66例糖尿病病人为单纯糖尿病组,另选取同期健康体检者60例为对照组。收集病人的一般临床资料并比较,采用尿素酶法检测血清尿素(UREA),氧化酶法检测肌酐(Cr)和尿酸(UA)水平,采用酶联免疫吸附法(ELISA)检测血清RNLS、UMOD、suPAR的水平。Pearson相关性分析RNLS、UMOD、suPAR与UREA、Cr、UA以及临床资料的关系。采用受试者操作特征(ROC)曲线分析血清RNLS、UMOD、suPAR联合检测对糖尿病病人发生肾损伤的诊断价值。结果糖尿病肾损伤组、单纯糖尿病组三酰甘油、总胆固醇、空腹血糖、糖化血红蛋白及血清UREA、Cr、UA、suPAR水平[(133.56±42.68)ng/L,(66.48±17.13)ng/L比(34.15±8.26)ng/L]均显著高于对照组(P<0.05),估算肾小球滤过率(eGFR)及血清RNLS[(28.62±7.61)mg/L,(35.47±9.29)mg/L比(42.81±13.24)mg/L]、UMOD水平[(64.28±16.82)g/L,(119.45±28.65)g/L比(139.82±32.84)g/L]均显著低于对照组(P<0.05);且糖尿病肾损伤组较单纯糖尿病组升高或下降幅度大(P<0.05)。糖尿病肾损伤组糖尿病病程显著高于单纯糖尿病组(P<0.05),24 h尿蛋白显著高于单纯糖尿病组和对照组(P<0.05)。临床期糖尿病肾病组病人血清RNLS[(26.28±6.69)mg/L比(31.18±8.34)mg/L]、UMOD水平[(51.26±14.41)g/L比(78.65±19.49)g/L]较早期糖尿病肾病组均显著降低(P<0.05),suPAR水平[(151.96±48.03)ng/L比(113.27±36.81)ng/L]显著升高(P<0.05)。RNLS、UMOD与UREA、Cr、UA、三酰甘油、总胆固醇、空腹血糖、糖化血红蛋白、糖尿病病程及24 h尿蛋白均呈负相关(P<0.05),与eGFR呈正相关(P<0.05);suPAR与UREA、Cr、UA、三酰甘油、总胆固醇、空腹血糖、糖化血红蛋白、糖尿病病程及24 h尿蛋白均呈正相关(P<0.05),与eGFR呈负相关(P<0.05)。ROC曲线分析结果显示,血清RNLS、UMOD、suPAR三项联合的曲线下面积(AUC)0.88显著高于RNLS、UMOD单项检测的AUC(0.81、0.83)(P<0.05),而与suPAR AUC比较差异无统计学意义(P>0.05)。结论糖尿病肾损伤病人血清RNLS、UMOD、suPAR水平呈异常表达,且三项联合检测对糖尿病病人发生肾损伤的诊断价值较高,值得临床推广应用。

Expression and significance of toll-like receptor 2,4 in peripheral blood mononuclear cells in patients with systemic inflammatory response syndrome

To explore changes of toll-like receptor (TLR) 2,4 in peripheral blood mononuclear cells (PBMC) in acute abdomen patients with systemic inflammatory response syndrome (SIRS) and their significance.Methods A clinical study was done on 103 patients of which 65 were with SIRS.The mRNA expression of TLR2,4 were detected by RT-PCR;the expression of TNF-α and IL-6 were observed by ELISA;the correlation between TLR2,4 mRNA,the level of TNF-α and IL-6,and the clinical course was evaluated.Results TLR2 mRNA ,TNF-α and IL-6 were upregulated markedly on the first day of hospitalization,then decreased gradually;TLR2 mRNA maintained on high level till the 5th day.The expression of TLR2,4 mRNA was positive correlated with the level of TNF-α and IL-6,and the length of stay.TLR2,4 mRNA expression increased in patients with multiple organ failure.Conclusion In actue abdomen patients with SIRS,the expression of TLR2,4 of PBMC increased markedly,indicating its improtant role in the pathogenesis of SIRS.4 refs,2 figs,2 tabs.

白细胞介素1受体颉颃剂抑制脂多糖促奶牛外周血单个核细胞氧化应激损伤作用的研究

试验以脂多糖(LPS)为刺激源,以细胞活力、抗氧化指标和炎症因子为判断指标,探讨白细胞介素1受体颉颃剂(IL-1Ra)通过抑制白细胞介素1β(IL-1β)的活性,对LPS诱导外周血单个核细胞(Peripheral blood mononuclear cells,PBMCs)氧化损伤的缓解作用。试验采用单因子完全随机设计,PBMCs被随机分为7个组(每组6个重复),分别给予不同的处理:第1组是阴性对照组(Neg组),完全培养基培养30 h;第2组损伤组(Dam组),是在完全培养基中培养6 h后,再经10μg/mL的LPS工作液培养24 h;第3至7组(R0.25、R0.5、R1、R5组和R10组)细胞分别经浓度为0.25、0.5、1、5、10 ng/mL的IL-1Ra培养6 h,接着经10μg/mL的LPS工作液培养24 h。结果表明:与Neg组相比,Dam组的细胞活力、抗氧化相关酶[包括总抗氧化能力(T-AOC)以及总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)和硫氧还蛋白还原酶(TrxR)]的活性显著降低,丙二醛(MDA)浓度、炎症因子白细胞介素-6(IL-6)和IL-1β含量以及诱导型一氧化氮合酶(iNOS)活性、一氧化氮(NO)含量均显著升高(P≤0.05)。与Dam组相比,R1组显著逆转了氧化损伤引起的上述抗氧化活性的降低和炎症因子浓度的升高,其他IL-1Ra处理组对上述指标的逆转效果不同程度地低于R1组(P≤0.05)。上述结果说明,LPS通过诱发PBMCs产生大量IL-1β进而导致细胞氧化损伤,IL-1Ra剂量依赖性地缓解了LPS引起的氧化损伤,添加剂量以1 ng/mL为宜。

金水宝联合胰高血糖素样肽-1受体激动剂治疗对糖尿病肾病患者肾功能、肾脏血流及血管内皮功能的影响

目的 探究金水宝联合胰高血糖素样肽-1受体激动剂(glucagon-likepeptide-1,LP-1)治疗对糖尿病肾病患者肾功能、肾脏血流及血管内皮功能的影响。方法 采用随机数字表法将2020年3月—2022年8月收治的96例糖尿病肾病患者分为观察组(n=48)与对照组(n=48)。对照组采用GLP-1受体激动剂,观察组在此基础上联合使用金水宝治疗。观察两组患者治疗前后中医证候积分、肾功能、肾脏血、氧化应激指标及血管内皮功能水平。观察两组患者临床疗效和不良反应发生率。结果 两组患者治疗前尿频、精神萎靡、口渴喜饮和易饥多食中医证候积分差异无统计学意义(P>0.05)。治疗后,观察组尿频、精神萎靡、口渴喜饮和易饥多食中医证候积分显著低于对照组(P<0.05)。两组患者治疗前24 h尿蛋白定量、同型半胱氨酸(homocysteine, Hcy)、尿白蛋白排泄率(urinary albumin excretion, UAER)、血清肌酐(serum creatinine, Scr)水平差异无统计学意义(P>0.05)。治疗后,观察组24 h尿蛋白定量、Hcy、UAER和Scr水平显著低于对照组(P<0.05)。两组患者治疗前段间动脉血流灌注指数(Perfusion Index, PI)、段间动脉胰岛素(Regular Insulin, RI)和叶间动脉RI水平差异无统计学意义(P>0.05)。治疗后,观察组段间动脉PI、段间动脉RI和叶间动脉RI水平显著低于对照组(P<0.05)。两组患者治疗前丙二醛(malondialdehyde;malonic dialdehyde, MDA)、脂质过氧化氢(lipid peroxidation, LPO)、谷胱甘肽过氧化物酶(glutathione Peroxidase, GPX)、总抗氧化能力(total antioxidative capability, T-AOC)水平差异无统计学意义(P>0.05)。治疗后,观察组MDA、LPO水平显著低于对照组,且GPX和T-AOC水平显著高于对照组(P<0.05)。观察组总有效率为89.58%(43/48)显著高于对照组72.92%(35/48)(χ^(2)=4.376,P=0.036)。观察组总不良反应发生率为8.32%(4/48),略低于对照组的10.42%(5/48),但差异无统计学意义(χ^(2)=0.123,P=0.726)。结论 金水宝联合GLP-1受体激动剂能有效提高临床疗效,改善肾功能和肾脏血流,促进血管内皮功能恢复。

三黄益肾胶囊调控NF-κB信号通路减轻糖尿病肾病大鼠炎症反应的机制研究

目的:旨在探究三黄益肾胶囊治疗糖尿病肾病(DN)可能的炎症反应机制。方法:按照随机数字表法将60只大鼠分为正常组、模型组、厄贝沙坦组及三黄益肾低、中、高剂量组,每组10只。除正常组外,其余各组大鼠用高糖高脂饲料联合链脲佐菌素(STZ)注射建立DN模型。正常组和模型组给予生理盐水2 mL灌胃;厄贝沙坦组给予厄贝沙坦11.51 mg/kg灌胃;三黄益肾低、中、高剂量组分别给予三黄益肾胶囊0.41、0.81、1.62 g/kg灌胃。各组大鼠均1次/d,连续给药4周。比较各组大鼠空腹血糖(FBG)、体质量、24 h尿白蛋白、肾功能指标、肾组织病理学变化、肾组织抗氧化相关指标(SOD、GSH-Px活性及MDA水平)、肾组织炎症介质[白细胞介素6(IL-6)、白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)]、肾组织诱导型一氧化氮合酶(iNOS)、肾组织CD68、CC趋化因子受体2(CCR2)表达、肾组织CC趋化因子配体2(CCL2)、NF-κB P65、p-NF-κB P65、IκB、p-IκB表达。结果:与模型组比较,各三黄益肾胶囊组大鼠体质量高,FBG、血清肌酐(Cr)、尿素氮(BUN)及24 h尿蛋白水平低,肾脏组织病理变化减轻,三肾组织中SOD、GSH-Px活性高,MDA水平低,IL-6、IL-1β、TNF-α水平低,肾组织中iNOS水平低,肾组织CCR2与CD68共定位水平低,CCL2、p-NF-κB P65、p-IκB蛋白表达低。结论:三黄益肾胶囊治疗DN的机制可能与抑制肾组织中NF-κB信号通路激活,减轻肾脏氧化应激及炎症反应有关。

基于我国WS/T促甲状腺激素受体抗体检测方法的性能验证

目的 基于我国卫生行业标准(WS/T)对迈瑞高敏磁微粒化学发光法(CLIA)检测TRAb进行性能验证。方法 参照最新WS/T文件,以Roche Cobase 601电化学发光法(ECLIA)作为已验证的检测方法,对迈瑞CLIA的精密度、基于患者血清的正确度、线性、检出限、参考区间进行验证,评价迈瑞与Roche的诊断符合率。结果 迈瑞CLIA检测TRAb的批内CV为1.56%~4.60%,实验室内CV为1.65%~5.19%;与Roche ECLIA的实验室相对差值为-3.6%;在0.02~44.00IU/L范围内线性拟合方程Y=1.040X+0.636,线性系数R2=0.990;24个检出限临界值数据22个(91.7%)≤0.3IU/L;20个健康体检者血清TRAb检测结果仅1个超出参考区间≤1.9IU/L;与Roche ECLIA的诊断总符合率90%,阳性符合率75%,阴性符合率100%,Kappa检验值0.7826(P<0.001)。结论 迈瑞高敏磁微粒CLIA的精密度、基于患者血清的正确度、线性范围、检出限均符合厂家声明,参考区间经验证可转移至我实验室使用,与Roche的诊断符合率较好,该方法性能能满足本医院临床检验需求。

红细胞膜表面TLR9表达在脓毒症相关性贫血中的临床价值

目的 探讨脓毒症患者外周血红细胞(RBC)膜表面Toll样受体(TLR)9表达在脓毒症相关性贫血中的临床价值。方法 选取2022年7月—2023年5月四川大学华西医院脓毒症患者237例(脓毒症组)、普通感染患者241例(普通感染组)和健康体检者206名(正常对照组)。收集所有研究对象的临床资料和脓毒症诊断24 h内的19项RBC相关参数检测结果。检测46例脓毒症患者、35例普通感染患者和28名健康体检者外周血RBC膜表面TLR9的表达情况。采用Spearman相关分析评估TLR9表达与RBC相关参数之间的相关性。结果脓毒症组RBC计数、血红蛋白(Hb)、平均红细胞血红蛋白浓度(MCHC)、网织红细胞绝对数(RET#)、网织红细胞血红蛋白(RET-He)、网织通道红细胞计数(RBC-O)、网织通道RET区域前向散射光强度(RET-Y)均显著低于普通感染组(P<0.05),红细胞平均体积(MCV)、红细胞体积分布宽度-标准差(RDW-SD)、红细胞体积分布宽度-变异系数(RDW-CV)、高荧光强度网织红细胞比例(HFR)均显著高于普通感染组(P<0.05)。脓毒症组TLR9+RBC百分比高于普通感染组(P<0.01)。TLR9+RBC百分比与RBC计数、Hb、RET#、RBC-O、MCHC、RET-He、RET-Y呈负相关(P<0.01),与RDW-SD、RDW-CV呈正相关(P<0.001)。结论 脓毒症患者RBC膜表面TLR9表达增加。高表达的TLR9与严重的贫血程度、低下的红系造血能力和异常的红细胞形态密切相关。

基于EGFR/MAPK/ERK信号通路探讨鳖甲煎丸对MHCC-97H肝癌细胞皮下瘤的抑瘤作用

目的基于表皮生长因子受体(epidermal growth factior receptor,EGFR)/丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)/细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)信号通路探究鳖甲煎丸对MHCC-97H肝癌细胞皮下瘤的抑瘤作用及作用机制。方法选取30只雄性BLAB/c裸鼠,建立MHCC-97H肝癌细胞皮下瘤模型。造模成功后随机分为模型组,鳖甲煎丸低、中、高剂量组(0.55、1.1、2.2 g/kg),西药组(乐伐替尼4 mg/kg+吉非替尼80 mg/kg),每组6只。鳖甲煎丸低、中、高剂量组灌胃2次/d,西药组每周灌胃5 d,模型组予以等量生理盐水2次/d灌胃,每组连续干预2周。观察大鼠一般情况;计算各组大鼠抑瘤率;HE染色观察病理形态学变化;RT-qPCR检测瘤体组织中EGFR、丝裂原活化蛋白质激酶激酶(mitogen-activated protein kinase kinase,MEK)、ERK1、ERK2 mRNA表达水平;Western blot检测EGFR、磷酸化的EGFR(p-EGFR)、MEK、磷酸化的MEK(p-MEK)、ERK1、ERK2、磷酸化的ERK1/2(p-ERK1/2)、基质金属蛋白酶-1(matrix metalloproteinase-1,MMP-1)、细胞周期蛋白D1(cell cycle protein D1,Cyclin D1)、神经型钙黏附蛋白(N-cadherin)、上皮型钙黏附蛋白(E-cadherin)相对表达水平。结果与模型组比较,鳖甲煎丸低、中、高剂量组及西药组精神、反应、进食饮水等情况均明显改善。与第0天比较,各组第14天体质量明显降低(P<0.01)。与模型组、鳖甲煎丸低剂量组比较,鳖甲煎丸中、高剂量组和西药组瘤体质量减轻(P<0.05,P<0.01)。鳖甲煎丸低、中、高剂量组和西药组抑瘤率分别为20%、47.73%、55.91%、75.45%。与模型组比较,鳖甲煎丸低、中、高剂量组及西药组肿瘤细胞排列疏松,边界模糊,细胞核固缩、破裂,其中西药组最明显。与模型组比较,鳖甲煎丸低、中、高剂量组和西药组EGFR、MEK、ERK1、ERK2 mRNA表达水平明显下降(P<0.01);与鳖甲煎丸低剂量组比较,鳖甲煎丸高剂量组和西药组EGFR、MEK、ERK1、ERK2 mRNA表达水平显著降低(P<0.01),鳖甲煎丸中剂量组ERK1 mRNA表达水平显著降低(P<0.01);与鳖甲煎丸中剂量组比较,西药组EGFR、ERK2 mRNA表达水平降低(P<0.05,P<0.01)。与模型组比较,鳖甲煎丸中、高剂量组和西药组p-EGFR/EGFR、p-MEK/MEK、p-ERK1/ERK1、p-ERK2/ERK2、MMP-1、Cyclin D1、N-cadherin蛋白相对表达水平下降(P<0.05,P<0.01),E-cadherin蛋白相对表达水平明显升高(P<0.01)。与鳖甲煎丸高剂量组比较,西药组p-EGFR/EGFR、p-ERK1/ERK1、MMP-1、Cyclin D1、N-cadherin蛋白相对表达水平明显下降(P<0.01),E-cadherin蛋白相对表达水平升高(P<0.05)。结论鳖甲煎丸可能通过抑制EGFR/MAPK/ERK信号通路激活,从而下调MMP-1、Cyclin D1、N-cadherin蛋白,上调E-cadherin蛋白表达,进而对MHCC-97H肝癌细胞皮下瘤产生显著的抑制作用。