Expression profile of polyunsaturated fatty acids in colorectal cancer

AIM:To investigate the relationship between the metabolism of polyunsaturated fatty acids(PUFAs)andtumor-associated factors for predicting the outcome of colorectal carcinoma(CRC)in Chinese patients.METHODS:Fresh-frozen malignant and normal tissues from 82 Chinese patients with CRC were analyzed for PUFA composition using gas-liquid chromatography.The levels of vascular endothelial growth factor(VEGF),cyclooxygenase-2(COX-2),prostaglandin E2 and platelet-derived growth factor(PDGF)were measured by enzyme-linked immunosorbent assay,and the levels of VEGF,p53 and Ki-67 were measured by immunohistochemistry.RESULTS:In malignant tissue,compared with normal tissue,the levels of totalω-6 PUFAs(24.64%±3.41%vs 26.77%±3.37%,P=0.00)and linoleic acid(LA)(15.46%±3.51%vs 18.30%±2.83%,P0.01)were lower,whereas the levels of totalω-3 PUFAs(1.58%±0.74%vs 1.35%±0.60%,P0.01)and dihomo-gamma-linolenic acid(DGLA)(1.32%±0.69%vs 0.85%±0.29%,P0.01)were significantly higher.The ratios of arachidonic acid(AA)/LA(0.53±0.22 vs0.42±0.19,P0.01)and AA/totalω-6 PUFAs(0.31±0.09 vs 0.27±0.10,P0.01)were also significantly higher in malignant tissue.The levels of PDGF(353.10±148.85 pg/m L vs 286.09±104.91 pg/m L,P0.01),COX-2(125.21±70.29 ng/m L vs 67.06±42.22 ng/m L,P0.01)and VEGF(357.11±128.76 pg/m L vs211.38±99.47 pg/m L,P0.01)were also higher in malignant tissue compared to normal tissue.COX-2was inversely correlated with LA(R=-0.3244,P0.05)and positively correlated with AA/totalω-6 PUFAs(R=0.3083,P0.05)and AA/LA(R=0.3001,P0.05).The tissue level of LA was highest in poorly differentiated tumors(19.9%±6.3%,P0.05),while the ratio of AA/ω-3 PUFAs was lowest in these tumors(10.8±2.6,P0.05).In VEGF-positive tumors,the level of LA was higher(16.2%±3.7%vs 13.9%±2.7%,P0.01),while the AA/ω-3PUFA,AA/ω-6 PUFA,and AA/LA ratios were lower than in VEGF-negativetumors(5.0±1.8 vs 6.7±3.3,0.30±0.09 vs 0.34±0.09,0.50±0.21 vs 0.61±0.21,P0.01).CONCLUSION:The metabolism of PUFAs may playan important role in the evolution of inflammationdriven tumorigenesis in CRC and may be considered apotential marker for prognosis.     

蛋白酶体抑制剂MG132抑制糖尿病肾脏疾病机制的研究

目的探讨蛋白酶体抑制剂MG132是否抑制糖尿病肾脏疾病(DKD)的发展及其可能的机制。方法 STZ复制糖尿病大鼠,随机分为糖尿病组(DM组,n=9)、MG132治疗组(MG132组,n=9),MG132组从第9周起予MG 132[0.1mg/(kg·d)]腹腔注射治疗糖尿病大鼠。同时设对照(Con组,n=9)组,检测生化指标,观察肾组织病理改变;Western blot检测肾组织SnoN、第10号染色体缺失的磷酸酶和张力蛋白同源基因(PTEN)、钙黏蛋白(E-cadherin)和α-平滑肌肌动蛋白(α-SMA)的表达。结果与Con组比较,DM组血糖[(5.85±0.86)vs(17.49±1.21)mmol/L,P=0.00]、尿微量白蛋白/尿肌酐比值(UACR)[(1.11±0.29)vs(16.36±3.06)mg/mmol,P=0.00]、肾脏指数(KW/BW)[(6.32±0.49)vs(14.54±1.49)mg/g,P=0.00]明显增加,且肾小管间质纤维化病变明显,α-SMA[(0.18±0.03)vs(0.33±0.02),P=0.002)增多,而SnoN[(0.87±0.10)vs(0.32±0.11),P=0.007)、PTEN[(2.06±0.09)vs(1.26±0.06),P=0.00]、E-cadherin[(1.32±0.06)vs(0.50±0.03),P=0.00]减少;MG132治疗后,UACR[(6.74±3.47)mg/mmol,P=0.003]、KW/BW[(12.43±1.11)mg/g,P=0.01]降低,纤维化病变减轻,α-SMA[(0.19±0.05),P=0.003]减少,SnoN[(0.55±0.09),P=0.033]、PTEN[(1.73±0.15),P=0.002]、E-cadherin[(1.11±0.10),P=0.00]蛋白的表达增加。结论 MG132可能通过恢复SnoN、PTEN蛋白水平抑制DKD的发展。     

RNA干扰Gal-3表达抑制胰腺癌细胞增殖并促进其凋亡

目的探讨RNA干扰半乳糖凝集素-3(galectin-3,Gal-3)表达对胰腺癌细胞增殖和凋亡的影响及其机制.方法将培养的Panc-1细胞随机分为对照组(未处理)、NC组(转染control-si RNA)和Gal-3干扰组(转染Gal-3-si RNA),以小干扰RNA(Small interfering RNA,si RNA)技术干扰Panc-1细胞中Gal-3表达后,RT-PCR和Western blot检测干扰效果,CCK-8法检测细胞增殖,流式细胞仪检测细胞凋亡,Western blot检测细胞中ki67、细胞周期蛋白D1(Cyclin D1)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved caspase-3)抗体和β-连环蛋白(β-catenin)蛋白表达.结果与对照组相比,NC组中Gal-3 m RNA(0.99±0.08vs 1.01±0.06)和蛋白(0.36±0.03 vs 0.34±0.05)的表达差异不显著(P0.05),而Gal-3干扰组中Gal-3m RNA(0.38±0.02 vs 1.01±0.06)和蛋白(0.10±0.01vs 0.34±0.05)的表达均显著降低(P0.05);Gal-3干扰组细胞增殖能力减弱(24 h:0.55±0.03 vs 0.71±0.05;48 h:0.76±0.05 vs 0.97±0.06;72 h:1.08±0.06 vs1.32±0.09),G0/G1期细胞百分比升高(79.48±1.32 vs71.52±1.15),S期(14.26±1.08 vs 18.24±1.06)和G2/M期(6.21±0.78 vs 10.19±1.52)降低,凋亡能力增强(13.26±2.28 vs 5.82±0.35),与对照组相比,差异均有统计学意义(P0.05);与对照组相比,Gal-3干扰组中ki67(0.24±0.02 vs 0.96±0.07)、CyclinD 1(0.26±0.03vs 0.88±0.09)和β-catenin(0.42±0.05 vs 0.87±0.05)蛋白的表达水平均明显降低,而Cleaved Caspase-3(0.70±0.06 vs 0.32±0.03)蛋白表达水平明显升高,差异有统计学意义(P0.05);NC组与对照组间各指标差异均无统计学意义(P0.05).结论 RNA干扰Panc-1细胞中Gal-3表达,可抑制细胞增殖并促进细胞凋亡,其机制可能与抑制Wnt/β-catenin信号通路有关.     

系统性红斑狼疮并发蛋白丢失性肠病的危险因素

目的对系统性红斑狼疮(systemic lupus erythematosus,SLE)并发蛋白丢失性肠病(protein-losing enteropathy,PLE)进行病例-对照研究,旨在总结本病的临床表现特点,探讨该并发症的危险因素。方法回顾性分析2000至2015年在北京协和医院住院的60例SLE并发PLE患者(PLE组)的临床资料,并与同期120例活动性SLE患者(对照组)进行比较。采用多因素Logistic回归分析方法,分析实验室各指标与并发PLE的关系,找出其危险因素。结果 PLE组患者平均血清白蛋白、补体C3、补体C4、血钙和24 h尿蛋白水平均显著低于对照组[(17.3±0.68)g/L vs.(32.3±0.80)g/L,P0.001;(0.47±0.03)g/L vs.(0.62±0.03)g/L,P=0.001;(0.08±0.017)g/L vs.(0.09±0.018)g/L,P=0.622;(1.84±0.02)mmol/L vs.(2.08±0.02)mmol/L,P0.001;(0.88±0.34)g/24 h vs.(2.76±0.36)g/24 h,P0.001],而血胆固醇水平显著高于对照组[(7.36±0.36)mmol/L vs.(5.41±0.24)mmol/L,P0.001]。PLE组患者血清抗SSA抗体(anti-SSA)和抗SSB抗体(anti-SSB)的阳性率高于对照组(61.7%vs.45.0%,P=0.035;23.3%vs.6.67%,P=0.001)。多因素Logistic回归分析显示,血清抗SSA抗体阳性、低白蛋白血症及高胆固醇血症是SLE并发PLE的独立危险因素(OR=3.520,P=0.024;OR=0.740,P0.001;OR=0.815,P=0.041)。结论 SLE患者出现血清抗SSA抗体阳性、高胆固醇血症、严重低白蛋白血症时,应警惕SLE并发PLE的发生。     

老年高脂血症患者补体C3、C4、备解素与踝臂指数的关系

目的 探讨老年高脂血症患者补体与踝臂指数(ABI)的关系.方法 选老年高脂血症患者257例,正常对照组60例,测定补体C3、C4、备解素(Pf)水平及ABI,分析其关系.结果 与对照组比较,高脂血症组C3 、C4 、Pf明显升高[(1.46 ±0.40)g/L vs (1.21 ±0.26) g/L,(0.37±0.14) g/L vs (0.29 ±0.09)g/L,(0.39±0.11)g/L vs (0.35±0.06)g/L,P=0.000,0.000,0.007];ABI降低[(1.12±0.15) vs(1.21±0.11),P=0.037],ABI与年龄(r=-0.243,P ＜0.05)、hsCRP(r=-0.330,P＜0.01)、性别(r=-0.284,P＜0.05)、TC(r=-0.276,P＜0.01)、收缩压(r=-0.228,P＜0.05)负相关,ABI与BMI正相关(r=0.257,P＜0.05),ABI与血清补体C3、C4、Pf不相关.结论 老年高脂血症患者ABI减低,ABI减低与补体C3、C4、Pf无直接关系.     

恶性消化道肿瘤晚期患者血TNF-α与胰岛素抵抗和胰岛素分泌的功能关系

目的:证明恶性消化道肿瘤晚期患者血浆肿瘤坏死因子α(TNF-α)与胰岛素抵抗和胰岛素分泌功能相关性,阐明恶性消化道肿瘤晚期患者糖代谢障碍机制.方法:测定40例恶性消化道肿瘤晚期患者和40例正常人员血浆TNF-α和空腹血糖、胰岛素、乳酸含量,并计算稳态模式评估法胰岛素抵抗指数(HOMA-IR)和胰岛素分泌指数(HOMA-β).结果:恶性消化道肿瘤晚期患者血浆TNFα、空腹血糖、空腹胰岛素、空腹乳酸、HOMA-IR和HOMA-β均明显高于正常对照组(3.27±0.92vs1.23±0.36,P＜0.01;5.19±0.75vs4.05±0.28,P＜0.01;14.24±6.52vs8.27±4.84,P＜0.01;7.11±0.69vs3.27±0.41,P＜0.01;3.48±0.85vs1.55±0.77,P＜0.01;181±39vs326±47,P＜0.01),但空腹血糖仍在正常参考范围内.恶性消化道肿瘤晚期患者血TNFα与空腹血糖、空腹胰岛素、空腹乳酸明显正相关(r=0.4352,P＜0.05;r=0.3136,P＜0.05;r=0.7893,P＜0.01),与HOMA-IR明显正相关(r=0.6531,P＜0.01),与HOMA-β明显负相关(r=-0.5874,P＜0.01).结论:恶性消化道肿瘤晚期患者血浆TNFα在胰岛素抵抗和胰岛素分泌功能下降及糖代谢障碍中发挥重要作用.     

Hsp22对缺氧/复氧损伤人脐静脉内皮细胞保护及Bcl-2、Caspase-3的表达

目的研究热休克蛋白22(Heat shock protein22,Hsp22)对缺氧/复氧损伤人脐静脉内皮细胞(HUVECs)保护作用,并探讨Hsp22调节bcl-2和caspase-3表达。方法把pGcsi-U6/Neo/GFp/shRNA/HSP22稳定转染HUVECs,Western blot印迹检测Hsp22蛋白在对照组、Lipo组、未转染组和转染组缺氧24 h/复氧0 h表达;MTT检测转染组和未转染组缺氧24 h/复氧0 h、3 h、6 h、12 h生长抑制:RT-PCR和Western blot印迹检测bcl-2和caspase-3转染组和未转染组缺氧24 h/复氧0 h、3 h、6 h、12 h基因和蛋白表达水平。结果 (1)转染组显著低于比对照组、Lipo2000组和未转染组(0.20±0.02 vs.0.43±0.03 vs.0.45±0.06 vs.0.48±0.05,P<0.05)而对照组、Lipo2000组和未转染组组间无显著差异(P>0.05)。(2)转染组抑制高于未转染组相同复氧时间点(58.5%±2.1%vs.41.6%±5.4%、62.7%±5.4%vs.45.6%±3.7%、65.4%±8.4%vs.48.5%±5.2%和68.6±6.7%vs.52.9%±3.4%,P<0.05)。(3)转染组bcl-2基因和蛋白在相同复氧时间点均低于未转染组:(1.85±0.06 vs.2.65±0.15、1.66±0.04 vs.2.35±0.08、1.09±0.05 vs.2.05±0.08、0.69±0.04 vs.1.75±0.09,P<0.05)和(0.54±0.04 vs.1.05±0.05、0.32±0.05 vs.0.75±0.03、0.29±0.02 vs.0.55±0.03、0.13±0.04 vs.0.45±0.07,P<0.05)。(4)转染组caspase-3基因和蛋白在相同复氧时间点均高于未转染组:(2.75±0.04 vs.1.75±0.07、2.96±0.04 vs.1.95±0.06、3.59±0.05 vs.2.45±0.05、3.59±0.05 vs.2.77±0.09,P<0.05)和(9.36±0.11 vs.3.05±0.12、8.82±0.07 vs.2.65±0.12、6.18±0.24 vs.2.05±0.11、3.59±0.05 vs.1.75±0.04,P<0.05)。结论 (1)稳定转染并缺氧/复氧损伤后,转染组Hsp22蛋白表达明显低于对照组而对照组、Lipo组和未转染组组间Hsp22蛋白无显著差异;(2)Hsp22对缺氧/复氧损伤HUVECs起保护作用;(3)Hsp22通过上调bcl-2和下调caspase-3基因和蛋白表达保护受损细胞。     

雷公藤甲素对2型糖尿病大鼠肾组织足细胞Nephrin、Podocin蛋白表达的影响及机制

目的 观察雷公藤甲素对2型糖尿病(T2DM)大鼠肾组织足细胞Nephrin、Podocin蛋白表达的影响及其机制探讨.方法 SPF级雄性8周龄Wistar大鼠50只,体重(200±20)g,按随机数字表法分为2组:对照组10只、模型组40只.模型组高脂高糖喂养8周联合小剂量链脲佐菌素(STZ)30mg/kg建立T2DM模型鼠,正常组大鼠喂养常规饲料.将造模成功的T2DM大鼠(28只)随机分成2组:糖尿病组(14只)和雷公藤甲素治疗组(14只),雷公藤甲素治疗8周后,检测大鼠生化指标、肾重/体重和尿白蛋白.利用光镜、电镜观察肾脏病理改变.采用实时定量PCR、免疫组化及Western blot法检测肾组织Nephrin及Podocin蛋白、骨桥蛋白、转化生长因子-β1(TGF-β1)及单核/巨噬细胞表面特异性标志抗原(ED-1)的mRNA和蛋白表达分布.采用单因素方差分析法进行统计学分析.结果 糖尿病大鼠尿白蛋白明显高于对照组(F=181.51,P＜0.01),治疗组尿白蛋白明显低于DM组(F=181.51,P＜0.01).与对照组比较,糖尿病大鼠肾组织Nephrin(F=36.82,P＜0.05)和Podocin(F=32.57,P＜0.05)蛋白表达下调,Nephrin(F=88.45,P＜0.01)、Podocin(F=55.43,P＜0.01)mRNA表达下调;雷公藤甲素组干预8周后Nephrin[(1.82±0.06)和(1.63±0.06),P＜0.05]和Podocin[(1.80±0.06)和(1.60±0.06),P＜0.05]蛋白质表达上调,肾脏组织Nephrin[(0.73±0.12)和(0.19±0.08),P＜0.01]和Podocin[(0.60±0.12)和(0.26±0.07),P＜0.01]mRNA表达上调.与对照组比较,T2DM大鼠肾组织骨桥蛋白(F=40.06,P＜0.05)和TGF-β1的(F=28.84,P＜0.05)蛋白质表达上调,肾组织骨桥蛋白(F=177.46,P＜0.01)和TGF-β1(F=161.27,P＜0.01)mRNA表达上调,雷公藤甲素治疗后肾组织骨桥蛋白[(1.27±0.03)和(1.39±0.05),P＜0.05]和TGF-β1[(1.23±0.03)和(1.44±0.02),P＜0.05]的蛋白表达下调,肾组织骨桥蛋白[(2.05±0.16)和(3.26±0.26),P＜0.01]和TGF-β1[(4.0±0.70)和(6.5±0.71),P＜0.01]mRNA表达下调.结论 雷公藤甲素对T2DM大鼠足细胞有保护作用,其机制可能与其?     

Protective effects of D-002 on experimentally induced gastroesophageal reflux in rats

AIM:To investigate the effects of beeswax alcohols(D-002)on the esophageal damage induced by gastroesophageal reflux(GER)in rats.METHODS:Sixty male rats were randomized into six groups(10 rats/group):a negative control and five groups with experimentally induced GER:a positive vehicle control,three treated with D-002(25,100 and 200mg/kg,respectively),and one with omeprazole 10 mg/kg.All treatments were given by gastric gavage.One hour after dosing,GER was produced by simultaneous ligation of the pyloric end and the forestomach.Esophageal lesions index(ELI),gastric secretion volume and acidity,and esophageal malondialdehyde(MDA)and sulfhydryl(SH)group concentrations were measured.Statistical significance was considered at P<0.05.RESULTS:As compared to the negative control,the positive control group exhibited increased ELI(5.2±0.33 vs 0±0,P=0.0003),gastric secretion volume(2.69±0.09 vs 0.1±0.0,P=0.0003)and acidity(238±19.37 vs 120.0±5.77,P=0.001),and esophageal concentrations of MDA(2.56±0.1 vs 1.76±0.28,P=0.001)and SH groups(1.02±0.05 vs 0.56±0.08,P=0.0003).D-002(25,100 and 200 mg/kg)reduced ELI(3.36±0.31,2.90±0.46 and 2.8±0.23,respectively)vs the positive control(5.2±0.33)(P=0.004;P=0.002;P=0.001,respectively).There were no significant changes in acidity with D-002 treatment,and only the highest dose reduced the volume of the gastric secretion(1.92±0.25)vs the positive control(2.69±0.09,P=0.013).D-002(25,100 and 200 mg/kg)lowered the esophageal MDA(2.05±0.16,1.98±0.22and 1.93±0.22,respectively)(P=0.01;P=0.03;P=0.03,respectively)and SH group concentration(0.87±0.06,0.79±0.08 and 0.77±0.06,respectively)(P=0.04;P=0.04;P=0.02)vs the positive control(2.56±0.10 and 1.02±0.05,respectively).Omeprazole decreased ELI(2.54±0.47),gastric secretion volume(1.97±0.14)and acidity(158.5±22.79),esophageal MDA(1.87±0.13)and SH group(0.72±0.05)concentrations vs the positive control(P=0.002;P=0.001;P=0.02;P=0.003;P=0.002,respectively).CONCLUSION:Acute oral administration of D-002 decreased macroscopic esophageal lesions and oxidative stress in rats with experimentally induced GER,without modifying gastric secretion acidity.