Effect of Electroacupuncture on Platelet-derived Growth Factor and the Ultrastructure of Mitochondria in Rats with Diabetic Gastroparesis

Objective To observe the effect of electroacupuncture(EA)at the pressure points Zu San Li(ST36),San Yin Jiao(SP6)and Liang Men(ST21)on platelet-derived growth factor(PDGF)and the ultrastructure of mitochondria in rats with diabetic gastroparesis(DGP).Methods Sixty Sprague Dawley(SD)rats were randomly separated into a normal control group(NC,n=10)and a modeling group(n=50).Rats in the modeling group received an injection of 2%streptozotocin(STZ)and a high-fat and highglucose diet for eight weeks to establish a DGP rat model.At the same time,blood glucose and a general symptom score were recorded every week.After modeling,30 successfully modeled rats were randomly separated into the following groups:the DGP group(n=10),the EA group(n=10)and the metoclopramide(MP)group(n=10).After three weeks of intervention,the gastrointestinal propulsive rate was measured by measuring the optical density(OD).The concentration of Ca2+was determined by fluorescence immunoassay,and levels of serum insulin(INS)and PDGF were determined by ELISA.The ultrastructure of mitochondria was observed with transmission electron microscopy.Results(1)After intervention,levels of blood glucose and the general symptom score were greatly decreased in the EA group compared to the DGP group(P<0.01).Compared with the DGP group,the gastric emptying rate and the intestinal propulsive rate of the EA group was significantly improved(P<0.01),and there was no statistically significant difference between the EA and the NC groups.(2)Compared with the NC group,the levels of INS in the DGP group markedly decreased(P<0.05),but there was no significant difference of INS levels between the EA and the MP roups.(3)Compared with the DGP group,theconcentration of Ca2+in the EA and the MP groups significantly increased(P<0.01,P<0.05,respectively).(4)Compared with the NC group,the average OD of PDGF in the DGP group was significantly higher(P<0.01).Compared with the DGP group,levels of PDGF in the EA group increased significantly(P<0.01).(5)There were abundant mitochondria with a clear structure and complete cristae in the NC group.However,in the DGP group,mitochondria were severely swollen,partly vacuolated,and cristae were either fractured,absent,or shortened.In the EA group,mitochondria were slightly swollen,with clear cristae.Conclusions Electroacupuncture at the points Zu San Li(ST36),San Yin Jiao(SP6)and Liang Men(ST21)may improve gastric motility in DGP by up-regulating the amount of PDGF and improving the ultrastructure of mitochondria.

CHF患者CRP/Alb、PDGF、BUN/Cr水平变化及对预后的评估价值

目的 探讨慢性心力衰竭(CHF)患者C-反应蛋白与白蛋白比值(CRP/Alb)、血小板源性生长因子(PDGF)联合血尿素氮与肌酐比值(BUN/Cr)水平变化,并分析各指标预测心血管死亡和全因死亡的效能。方法 选取2020年1月至2022年3月收治的109例CHF患者,根据1年内心血管死亡和全因死亡情况分为死亡组(26例)和存活组(83例)。比较2组社会人口学资料和临床特征[美国纽约心脏病学会心功能分级(NYHA)、左心室射血分数、左心室舒张末期内径、左心室收缩末期内径、N-末端脑钠肽前体(NT-proBNP)、临床用药]、CRP/Alb、PDGF、BUN/Cr水平,分析CRP/Alb、PDGF、BUN/Cr水平的相关性,采用Cox回归分析CHF患者心血管死亡和全因死亡的相关影响因素,采用受试者工作特征曲线(ROC)分析预测价值,采用卡普兰-迈耶曲线(K-M)生存曲线分析不同CRP/Alb、PDGF、BUN/Cr水平患者的生存率。结果 死亡组NYHA心功能分级、左心室舒张末期内径、左心室收缩末期内径、NT-proBNP高于存活组,左心室射血分数低于存活组(P<0.05);死亡组CRP/Alb、PDGF、BUN/Cr高于存活组(P<0.05);CHF患者CRP/Alb与PDGF、BUN/Cr呈正相关,PDGF与BUN/Cr呈正相关(P<0.05);在校正了NYHA心功能分级等混杂因素后,CRP/Alb、PDGF、BUN/Cr仍是CHF患者心血管死亡和全因死亡的相关影响因素(P<0.05);CRP/Alb、PDGF、BUN/Cr预测心血管死亡和全因死亡的ROC下面积(AUC)分别为0.712、0.801、0.738,三者联合时的AUC为0.911(P<0.05);CRP/Alb、PDGF、BUN/Cr高水平患者生存率低于低水平患者(P<0.05)。结论 CRP/Alb、PDGF、BUN/Cr与CHF患者心血管死亡和全因死亡有关,联合检测可作为预测患者病情转归的一个方案,为临床评估预后、治疗等提供参考依据。

基于核酸适体与互补核酸和目标蛋白之间竞争反应的PDGF蛋白特异性识别

报道了一种利用核酸适体与互补核酸和目标蛋白之间的竞争反应用于PDGF蛋白的特异性识别方法.采用反相微乳液技术制备了包覆有亚甲基蓝(MB)的SiO2纳米复合物(SiO2-MB),利用固定在磁性纳米颗粒上的核酸适体与SiO2-MB纳米颗粒标记的互补核酸进行杂交反应,将一定数量的SiO2-MB纳米颗粒固定于磁性纳米颗粒的表面,引入PDGF目标蛋白后,利用核酸适体与互补核酸和PDGF之间的竞争反应,通过检测MB电化学信号的变化来检测PDGF.该方法对PDGF蛋白具有很高的特异性识别能力,不受其它蛋白质如免疫球蛋白G、凝血酶和溶菌酶等的干扰.实验结果表明,PDGF浓度在5.51×10-17～1.01×10-15 mol/L范围内具有良好的线性关系,检出限可达1.31×10-17 mol/L.

Role of RhoA in platelet-derived growth factor-BB-induced migration of rat hepatic stellate cells

Background Although the migration of hepatic stellate cells （HSCs） is essential for hepatic fibrotic response, the detailed mechanisms involved are poorly understood. The aim of this study was to examine the role of Rho GTPases （especially RhoA） in platelet-derived growth factor （PDGF）-BB-induced migration of HSCs. Methods The migration of primary rat HSCs was evaluated using transwell Boyden chamber, while cytoskeletal changes were visualized by immunofluorescence staining of intracellular actins and vinculin. Quantitative real-time PCR and Western blotting analysis were used to detect the expression of Rho GTPases （RhoA, Racl and Cdc42） within HSCs and their activation was determined by glutathione S-transferase pull-down assay. Finally, the effects of RhoA on PDGF-BB-induced cell migration and cytoskeletal remodeling were analyzed using HSC-T6 cells stably transfected with constitutively active （CA, Q63L） or dominant negative （DN, T19N） RhoA mutants. Data were analyzed using SPSS 16.0 software. Student＇s t test was used to analyze differences between two groups and one-way analysis of variance （ANOVA） was used among multiple groups. Results Rapid cytoskeletal remodeling led to a significant increase in the motility of primary rat HSCs after haptotactic （direct） and chemotactic （indirect） stimulation by PDGF-BB： PDGF-BB caused a dramatic elevation in the levels of both total and active RhoA protein. However, the levels of mRNA for Rho GTPases, including RhoA, Racl and Cdc42, were unaffected. Furthermore, PDGF-BB induced increased formation of stress fibers and focal adhesions in HSC-T6 cells transfected with CA-RhoA, but not in HSC-T6 transfected with DN-RhoA. Surprisingly, both CA- and DN-RhoA-transfected HSC-T6 cells showed decreased migratory potential in the absence or presence of PDGF-BB compared with controls. Conclusions PDGF-BB induced cytoskeletal remodeling in rat HSCs and promoted their migration via regulation of intracellular RhoA. RhoA may be one of the determinants in PDGF-BB-induced HSC miaration.

The relationship between PDGF and angiogenesis in endometrial cancer

Objective:To investigate the expression of platelet-derived endothelial growth factor(PDGF) and its effect in angiogenesis in endometrial cancer(EC),in order to investigate the mechanism of tumorigenesis and lay a foundation for the management of EC.Methods:We selected 40 cases with EC,20 endometrium,and 20 normal endometrium.All specimens were stained immunohistochemically for CD34 and PDGF identified immunohistochemically with specific antibodies.Results:The expression of PDGF was significantly higher in EC than normal endometrium and atypical hyperplasia of endometrium(P < 0.05),however,no significant difference was found between normal endometrium and atypical hyperplasia of endometrium(P > 0.05).The expression of PDGF was lower in well differentiated than moderately and poorly differentiated EC(P < 0.05).In cases with muscular invasion tumors,it was higher than in normal endometrium(P < 0.05).After Spearman correlation analysis,MVD was significantly influenced by PDGF,r = 0.848(P = 0.000).Conclusion:There was positive correlation between microvessel density(MVD) and PDGF in earlier stage of EC,it illustrate that PDGF may take part in angiogenesis of EC.

复肾降纤宁对糖尿病肾病大鼠的影响及机制研究

目的观察复肾降纤宁对糖尿病肾病(DN)大鼠肾脏蛋白激酶C(PKC)、血管内皮生长因子(VEGF)和血小板衍生生长因子-β(PDGF-β)的影响及对肾脏的保护作用。方法建立链脲佐菌素(STZ)诱导的糖尿病肾病(DN)大鼠模型,将成模糖尿病大鼠随机分成4组:模型组、复肾降纤宁组、厄贝沙坦组、厄贝沙坦+复肾降纤宁组,同时取正常大鼠设对照组。各组大鼠采用相应的干预措施处理12周。常规方法检测各组大鼠肾脏指数、血糖、尿素氮(BUN)、血肌酐(Scr)、尿白蛋白排泄率(UAER),免疫组化法检测肾PKC和VEGF表达,Western blotting检测肾组织PDGF-β的表达,透射电镜观察肾脏超微结构。结果模型组大鼠肾皮质PKC、VEGF和PDGF-β表达显著升高,肾脏超微结构改变明显,血糖、UAER、BUN、Scr、肾脏指数显著增高;复肾降纤宁组、厄贝沙坦组和厄贝沙坦+复肾降纤宁组上述指标显著低于模型组(P<0.05),肾脏超微结构改变明显改善;厄贝沙坦+复肾降纤宁组各项指标改善明显优于模型组、复肾降纤宁组和厄贝沙坦组(P<0.05),但未达到对照组水平。结论复肾降纤宁对DN大鼠肾脏形态和功能有明显保护作用,其机制可能与减少肾组织中PKC、VEGF和PDGF-β的表达,减轻肾组织纤维化有关。

Inhibitory effects of rapamycin on the different stages of hepatic fibrosis

AIM: To investigate and compare the inhibitory effects of rapamycin in the different stages of liver fibrosis.

Mechanisms responsible for the inhibitory effects of epothilone B on scar formation after spinal cord injury

Scar formation after spinal cord injury is regarded as an obstacle to axonal regeneration and functional recovery.Epothilone B provides moderate microtubule stabilization and is mainly used for anti-tumor therapy.It also reduces scar tissue formation and promotes axonal regeneration after spinal cord injury.The aim of the present study was to investigate the effect and mechanism of the microtubule-stabilizing reagent epothilone B in decreasing fibrotic scarring through its action on pericytes after spinal cord injury.A rat model of spinal cord injury was established via dorsal complete transection at the T10 vertebra.The rats received an intraperitoneal injection of epothilone B（0.75 mg/kg） at 1 and 15 days post-injury in the epothilone B group or normal saline in the vehicle group.Neuron-glial antigen 2,platelet-derived growth factor receptor β,and fibronectin protein expression were dramatically lower in the epothilone B group than in the vehicle group,but β-tubulin protein expression was greater.Glial fibrillary acidic protein at the injury site was not affected by epothilone B treatment.The Basso,Beattie,and Bresnahan locomotor scores were significantly higher in the epothilone B group than in the vehicle group.The results of this study demonstrated that epothilone B reduced the number of pericytes,inhibited extracellular matrix formation,and suppressed scar formation after spinal cord injury.

胞外基质蛋白与血小板生长因子对气道平滑肌细胞的影响

目的研究层黏蛋白(laminin,LN)、纤维连接蛋白(fibronectin,FN)、I型胶原(collagen I,Col I)3种不同细胞外基质(extracelluar matrix,ECM)蛋白对血小板生长因子(platelet derived growth factor-BB,PDGF-BB)诱导的气道平滑肌细胞(airway smooth muscle cells,ASMCs)形态及收缩变化的影响。方法将ASMCs分别附着在LN、ColI、FN表衬的细胞培养皿中,并分成2组,分别在有或无PDGF-BB(10 mg/L)的无血清培养基中培养0～5 d。然后采用显微图像方法观测细胞形态和宽长比,光学磁微粒扭转细胞测量方法测量细胞受KCl或组胺(histamine)刺激时的收缩反应。结果在有PDGF-BB的培养条件下,ASMCs形态总体上变细变长,即细胞宽长比减小,但附着在LN上的ASMCs比附着在Col I或FN上的细胞宽长比相对要大。在无PDGF-BB的培养条件下,ASMCs对KCl刺激的收缩响应随培养时间增加而增加,但不受ECM蛋白成分的影响。而在有PDGF-BB的培养条件下,ASMCs对KCl或Histamine刺激的收缩响应总体上随培养时间呈下降趋势,但附着在LN上的ASMCs收缩响应下降程度相对较小。结论 ASMCs受PDGF-BB作用时,其形态和收缩性的变化与不同ECM蛋白成分有关,相对于Col I和FN,附着在LN的细胞形态和收缩性的变化较小。ECM蛋白成分对PDGF-BB诱导的ASMCs形态和收缩性变化的差异性影响,对于深入认识基质蛋白、炎症因子与气道平滑肌细胞的相互作用及其与哮喘病理生理机制的关系具有重要意义。

川陈皮素对糖尿病肾病大鼠的作用研究

目的构建糖尿病肾病大鼠模型,从整体水平观察川陈皮素对其的治疗作用,并从分子水平研究川陈皮素对丝氨酸蛋白酶家族B成员7(serpin family B member 7,Megsin)、血小板衍生生长因子-BB(platelet-derived growth factor-BB,PDGF-BB)、细胞外信号调节蛋白激酶1/2(extracellular signal-regulated protein kinase,ERK1/2)和Ⅳ型胶原(collagen IV)蛋白及mRNA表达的影响。方法清洁级雄性SD大鼠120只分为正常组、糖尿病肾病模型组、贝那普利组、川陈皮素低、中和高剂量组,高脂高糖饮食1月+一次性腹腔注射链尿佐菌素(streptozotocin,STZ)诱导糖尿病肾病大鼠模型,相应药物治疗6周后处死,收集尿液、血液和肾脏,检测血糖、血清和尿液中肌酐和β2-MG、小鼠肾重与体重比(KW/BW),同时检测肾脏炎性因子IL-1、IL-6和TNF-α含量,收集肾组织标本,聚合酶链式反应和蛋白印记法测定肾组织中Megsin、PDGF-BB、pERK1/2及collagen IV的表达。结果肾脏组织HE染色可见,正常组大鼠形态正常,模型组出现明显肾小球萎缩和硬化,与模型组相比,贝那普利组、川陈皮素低、中和高剂量组均明显好转,此外贝那普利组和川陈皮素高剂量组基本一致;与正常组相比,模型组大鼠血糖、KW/BW、尿液肾功能指标UREA和β2-MG、血液肾功能指标UREA和β2-MG、肾脏炎性因子IL-1、IL-6和TNF-α、肾脏组织中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA均明显升高(P<0.05),与模型组相比,贝那普利组、川陈皮素低、中和高剂量组血糖、KW/BW、尿液肾功能指标UREA和β2-MG、血液肾功能指标UREA和β2-MG、肾脏炎性因子IL-1、IL-6和TNF-α、肾脏组织中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA均降低(P<0.05),此外川陈皮素各组血糖均明显低于贝那普利组(P<0.05),川陈皮素高剂量组IL-1、IL-6和TNF-α低于贝那普利组(P<0.05),但KW/BW、尿液UREA和β2-MG、血液UREA和β2-MG、肾脏中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA和贝那普利组相比没有差异。结论川陈皮素对糖尿病肾病大鼠有明显的治疗作用,这与川陈皮素可调节肾脏中megsin、PDGF-BB、pERK1/2及collagen IV的表达有关。

Current Perspectives on Sunitinib Targeted Therapy for Tumors

This review highlights therapeutic agents from recent cancer therapeutic trials showing the greatest potential for further clinical use for sunitinib in the near future. In fact, sunitinib is one of multi-tyrosine kinase inhibitors;tyrosine kinases are enzymes, which transfer phosphate groups from ATP to the hydroxyl group of tyrosine residues on signal transduction molecules. Phosphorylation of signal transduction molecules, in turn, induces dramatic changes in tumor growth, including activation of angiogenesis and DNA synthesis. Therefore, sustain efforts have been directed for developing inhibitors for angiogenesis, which is the marginal process for tumor growth and development through targeting TKs. Almost if not all angiogenesis inhibitors target the vascular endothelial growth factor (VEGF) signaling pathway.

Silencing ofα-complex protein-2 reverses alcohol-and cytokine-induced fibrogenesis in hepatic stellate cells

Background and aim:a-complex protein-2(aCP2)encoded by the poly(rC)binding protein 2(PCBP2)gene is responsible for the accumulation of type I collagen in fibrotic livers.In this study,we silenced the PCBP2 gene using a small interfering RNA(siRNA)to reverse alcohol-and cytokine-induced profibrogenic effects on hepatic stellate cells(HSCs).Methods:Primary rat HSCs and the HSC-T6 cell line were used as fibrogenic models to mimic the initiation and perpetuation stages of fibrogenesis,respectively.We previously found that a PCBP2 siRNA,which efficiently silences expression of aCP2,reduces the stability of type I collagen mRNA.We investigated the effects of the PCBP2 siRNA on cell proliferation and migration.Expression of type I collagen in HSCs was analyzed by quantitative real-time PCR and western blotting.In addition,we evaluated the effects of the PCBP2 siRNA on apoptosis and the cell cycle.Results:PCBP2 siRNA reversed multiple alcohol-and cytokine-induced profibrogenic effects on primary rat HSCs and HSC-T6 cells.The PCBP2 siRNA also reversed alcohol-and cytokine-induced accumulation of type I collagen as well as cell proliferation and migration.Moreover,the combination of LY2109761,a transforming growth factor-b1 inhibitor,and the PCBP2 siRNA exerted a synergistic inhibitive effect on the accumulation of type I collagen in HSCs.Conclusions:Silencing of PCBP2 using siRNA could be a potential therapeutic strategy for alcoholic liver fibrosis.

Aggravated pulmonary injury after subarachnoid hemorrhage in PDGF-B^ret/ret mice

Background:Recent advances in surgical and neuroprotective strategies could effectively manage the pathophysiological progression of subarachnoid hemorrhage(SAH).However,pulmonary dysfunction frequently occurs in SAH patients with an increased risk of unsatisfactory outcomes.Based on the similar microvascular structures in the blood-air barrier and blood-brain barrier and possible brain-lung crosstalks,we believe that pericytes may be involved in both neurological and pulmonary dysfunction after SAH.Methods:In our experiments,platelet-derived growth factor B(PDGF-B)retention motif knockout(PDGF-Bret/ret)mice and adeno-associated virus PDGF-B were employed to show the involvement of pericyte deficiency and PDGF-B expression.Neurological score,SAH grade,hematoxylin-eosin staining,and PaO2/FiO2 ratio analysis were performed to evaluate the neurological deficits and pulmonary functions in endovascular perforation SAH models at 24 h after surgery,as well as western blotting and immunofluorescence staining for underlying molecular expressions.Results:We found that neonatal PDGF-Bret/ret mice exhibited pulmonary atelectasis 12 h after birth.Further investigation showed a decrease in PaO2/FiO2 and lung-specific surfactant proteins in adult PDGF-Bret/ret mice.These dysfunctions were much worse than those in wild-type mice at 24 h after SAH.PDGF-B overexpression alleviated pulmonary dysfunction after SAH.Conclusions:These results suggested pulmonary dysfunction after SAH and the pivotal role of PDGF-B signaling for the pathophysiological process and future therapeutic targets of pulmonary injury treatment after SAH.Further studies are needed for pathophysiological investigations and translational studies on pulmonary injuries after SAH.

G蛋白耦联受体激酶结合蛋白1通过调节细胞外调节激酶1/2的活性促进成骨细胞迁移

目的探索G蛋白耦联受体激酶结合蛋白1(GITI)在成骨细胞迁移中的作用,并分析其机理。方法通过Western blot方法检测GIT1蛋白在鼠的成骨细胞内的表达；用免疫荧光染色方法确定：在血小板衍生生长因子(PDGF)不刺激和刺激的条件下,GIT1和细胞外调节激酶1/2(ERK1/2)在成骨细胞内的位置；用共同免疫沉淀的方法测定GIT1和ERK1/2相互结合,并且用免疫荧光双染的方法确定这两种蛋白相互结合的位置；用包含GIT1-RNA发夹结构的腺病毒感染成骨细胞后,用免疫荧光染色方法确定磷酸化ERK1/2(pERK1/2)在成骨细胞内的位置,用划痕愈合法检测在PDGF刺激下的迁移能力。结果在成骨细胞内,PDGF刺激导致了GIT1和ERK1/2的相互结合,并且这种结合发生在成骨细胞的局部粘附内。包含GIT1-RNA发夹结构的腺病毒明显抑制了pERK1/2招募至成骨细胞局部粘附内以及PDGF所刺激的成骨细胞的迁移。结论在PDGF刺激下,GIT1招募pERK1/2至成骨细胞的局部粘附内,从而促进成骨细胞的迁移。

Sunitinib mesylate inhibits proliferation of human colonic stromal fibroblasts in vitro and in vivoSunitinib mesylate inhibits proliferation of human colonic stromal fibroblasts in vitro and in vivo

Objective： Cancer stromal fibroblasts are important members of the cancer microenvironment. In this study, we determined the effect of sunitinib, a small molecule tyrosine kinase inhibitor, on the primary human colonic fibroblasts. Methods： Cell cycle analysis and cell proliferation assays were performed to evaluate the inhibitory effect of sunitinib in vitro. Western-blot analysis was performed to evaluate variations in the levels of phosphorylated plateletderived growth factor receptor β （PDGFR-β）, Akt, and ERK proteins. Co-injection of SW620 cells and colonic fibreblasts in nude mice was employed to test anti-growth efficacy in vivo. Results： Sunitinib was found to effectively inhibit the growth of primary colonic fibroblasts. Low-dose sunitinib blocked the PDGF-BB-induced cell proliferation and PDGFR-β signaling. Co-injection of SW620 cells and colonic fibroblasts in nude mice generated greater tumor volumes than single injection of SW620 cells. Sunitinib treatment inhibited the SW620 cell＋colonic fibroblast tumor growth more effectively than treatment of 5-fluorouracil. Conclusions： Sunitinib mesylate inhibited the proliferation of primary human colonic fibroblasts through target-inhibited PDGFR signaling in vitro and in vivo.