119例非小细胞肺癌中CCR9的表达及其预后分析

目的：探讨 CC 族趋化因子受体9( C-C chemokine receptor 9, CCR9)在非小细胞肺癌( non-small cell lung cancer, NSCLC)中的表达及其预后判断的价值。方法应用免疫组化PV-9000两步法检测119例NSCLC及相应癌旁正常肺组织中CCR9蛋白的表达,并分析CCR9表达与临床病理特征及患者总生存率的关系。结果 CCR9在NSCLC中的阳性率(54.6%)明显高于癌旁正常肺组织(10．1%)( P <0．05);CCR9表达与 NSCLC 病理组织类型、淋巴结转移、p-TNM 分期有关( P <0．05)。 Kaplan-Meier生存分析显示肺癌中CCR9蛋白表达与患者术后总生存率呈负相关(Log-rank=9．917,P=0．002)。单因素生存分析显示,淋巴结转移、p-TNM分期、CCR9蛋白表达与NSCLC患者术后总生存率有关( P<0．05)。多因素生存分析显示,CCR9是NSCLC患者术后总生存率的独立预测因素(RR=0．447,95%CI：0．201~0．993,P<0．05)。结论 CCR9阳性患者预后较差,其可作为NSCLC患者预后判断的新生物学标志物。     

Chemokines CCL2, 3, 14 stimulate macrophage bone marrow homing,proliferation, and polarization in multiple myeloma

We previously showed that macrophages (MΦs) infiltrate the bone marrow (BM) of patients with myeloma and may play a role in drug resistance. This study analyzed chemokines expressed by myeloma BM that are responsible for recruiting monocytes to the tumor bed. We found that chemokines CCL3, CCL14, and CCL2 were highly expressed by myeloma and BM cells, and the levels of CCL14 and CCL3 in myeloma BM positively correlated with the percentage of BM-infiltrating MΦs. In vitro, these chemokines were responsible for chemoattracting human monocytes to tumor sites and in vivo for MΦ infiltration into myeloma-bearing BM in the 5TGM1 mouse model. Surprisingly, we also found that these chemokines stimulated MΦ in vitro proliferation induced by myeloma cells and in vivo in a human myeloma xenograft SCID mouse model. The chemokines also activated normal MΦ polarization and differentiation into myeloma-associated MΦs. Western blot analysis revealed that these chemokines promoted growth and survival signaling in MΦs via activating the PI3K/Akt and ERK MAPK pathways and c-myc expression. Thus, this study provides novel insight into the mechanism of MΦ infiltration of BM and also potential targets for improving the efficacy of chemotherapy in myeloma.     

CXCL10对βTC6细胞TLR4表达及细胞凋亡的影响

目的探讨CXC趋化因子配体10(CXCL10)对βTC6细胞Toll样受体4(TLR4)表达及细胞凋亡的影响。方法体外培养βTC6细胞,0.1~10.0ng/mL CXCL10分别干预βTC6细胞12,24,48h后,应用Westernblot检测TLR4表达情况,流式细胞术和DNA Ladder检测细胞凋亡情况。结果与对照组比较,CXCL10干预12h,10.0ng/mL干预组开始出现TLR4蛋白的表达水平上调(0.840±0.049,P<0.05);干预24h,1.0和10.0ng/mL干预组TLR4的表达水平上调[分别为(0.851±0.052)和(0.893±0.030),P<0.05];干预48h,1.0和10.0ng/mL干预组TLR4的表达进一步上调[分别为(0.876±0.046)和(0.923±0.027),P<0.05],且各干预浓度两两比较,差别均有统计学意义(P<0.05)。CXCL10干预βTC6细胞24h,仅10.0ng/mL干预组细胞出现DNA梯状条带;干预时间延长至48h,1.0和10.0ng/mL干预组均出现DNA梯状条带。流式细胞术显示,CXCL10诱导细胞凋亡呈浓度依赖性。结论长时间高浓度的CXCL10作用将导致TLR4表达显著上调,并诱导β细胞凋亡。     

CXCR4抑制剂对大鼠肺型氧中毒肺损伤的干预作用

目的：研究趋化因子受体4（chemokine receptor 4,CXCR4）特异性拮抗剂AMD3100在大鼠肺型氧中毒（ pulmonary oxygen intoxication）肺组织损伤中的干预作用。方法40只雄性SD大鼠随机分为常压空气PBS组、常压空气抑制剂组、纯氧暴露PBS组及纯氧暴露抑制剂组,每组10只。纯氧暴露PBS组及纯氧暴露抑制剂组利用0．23 MPa纯氧加压暴露,出舱后通过HE染色切片观察大鼠肺组织病理改变,ELISA法测定炎症因子TNF-α及IL-1β含量变化,应用Western印迹方法研究大鼠肺组织caspase-3含量变化。结果纯氧暴露抑制剂组大鼠肺组织淤血、水肿、出血与纯氧暴露PBS组相比程度较轻。纯氧暴露抑制剂组TNF-α、IL-1β及活化的caspase-3含量均低于纯氧暴露PBS组。结论应用AMD3100阻断CXCR4来减轻肺型氧中毒肺损伤是有效的。     

趋化因子诱饵受体D6在心肌梗死后心肌重塑中的作用及机制研究

目的:研究急性心肌梗死(AMI)后患者体内趋化因子诱饵受体D6的水平变化过程以及D6在心肌梗死后影响心肌重塑的可能机制。方法:将35例AMI患者根据心肌梗死后72 h血清趋化因子CCL2的检测水平分为h CCL2high组(n=18)和h CCL2low组(n=17),流式细胞术分析外周血中CD14++CD16-炎症单核细胞比例,Q-PCR测定外周血单核细胞趋化因子诱饵受体D6 mRNA的表达,3周后超声心动图测定心脏功能。将30只AMI雄性BLAB/C小白鼠根据血清CCL2的检测水平分为m CCL2high组(n=17)和m CCL2low组(n=13),流式细胞术分析小鼠外周血中CD11b+单核细胞比例;梗死心肌冰冻切片后免疫荧光染色D6阳性细胞,Q-PCR测定心肌梗死后3 d时间梗死区D6 mRNA的表达。3周后通过小动物超声检测小鼠心脏功能。结果:h CCL2high组患者外周血中CD14++CD16-炎症单核细胞比例显著高于h CCL2low组(P<0.001),其单核细胞D6受体的表达明显低于h CCL2low组(P<0.001),但单核细胞D6的表达与患者LVEF%未见明显相关性(P=0.16)。急性心肌梗死小鼠m CCL2high组外周血中CD11b+单核细胞细胞比例显著高于m CCL2low组(P<0.001),梗死心肌组织D6 mRNA表达显著低于m CCL2low组(P<0.001),小鼠梗死心肌免疫荧光可见D6表达于CD11b阳性单核细胞和CD31阳性内皮细胞。3周后小鼠心脏超声示LVEF%与梗死区D6的表达呈正相关(P=0.01)。结论:急性心肌梗死后梗死区组织可表达趋化因子诱饵受体D6,梗死区D6的高表达可抑制心肌梗死后的炎症浸润。梗死区趋化因子诱饵受体D6通过特异性结合CCL2减少心肌梗死后心肌局部的炎症浸润进而减轻心肌梗死后的心肌重塑。     

Circulating CCL20: A potential biomarker for active vitiligo together with the number of Th1/17 cells

Background

Vitiligo is an autoimmune disease with varying pathological features. Activation of the CCL20-CCR6 axis plays an important role in chronic inflammatory diseases. However, whether CCL20-CCR6 and Th1/17 cells are indicative of active vitiligo is unclear.

Decreased circulating CXCR3 + CCR9+T helper cells are associated with elevated levels of their ligands CXCL10 and CCL25 in the salivary gland of patients with Sjögren’s syndrome to facilitate their concerted migration

CCR9 + T helper (Th) cells can induce Sjögren-like symptoms in mice and both CCR9 + Th cells and their ligand CCL25 are increased in the salivary glands of primary Sjögren's syndrome (pSS) patients. Increased circulating CCR9 + Th cells are present in pSS patients. CCR9 + Th cells are hyperresponsive to IL-7, secrete high levels of IFN-γ, IL-21, IL-17 and IL-4 and potently stimulate B cells in both patients and healthy individuals. Our aim was to study co-expression of chemokine receptors on CCR9 + Th cells and whether in pSS this might differentially affect CCR9 + Th cell frequencies. Frequencies of circulating CCR9 + and CCR9- Th cells co-expressing CXCR3, CCR4, CCR6 and CCR10 were studied in pSS patients and healthy controls. CCL25, CXCL10, CCL17, CCL20 and CCL27 mRNA and protein expression of salivary gland tissue of pSS and non-Sjögren's sicca (non-SS) patients was assessed. Chemotaxis assays were performed to study migration induced by CXCL10 and CCL25. Higher expression of CXCR3, CCR4 and CCR6 but not CCR10 was observed on CCR9 + Th cells as compared to cells lacking CCR9. Decreased frequencies of circulating memory CCR9 + CXCR3+ Th cells were found in pSS patients, which was most pronounced in the effector memory subset. Increased salivary gland CCL25 and CXCL10 expression significantly correlated and both ligands functioned synergistically based on in vitro induced chemotaxis. Decreased memory CXCR3 + CCR9+ Th cells in blood of pSS patients may be due to a concerted action of overexpressed ligands at the site of inflammation in the salivary glands facilitating their preferential migration and positioning in the lymphocytic infiltrates.     

Low-dose naltrexone inhibits the epithelial-mesenchymal transition of cervical cancer cells in vitro and effects indirectly on tumor-associated macrophages in vivo

The metastasis of cervical cancer has always been a clinical challenge. We investigated the effects of low-dose naltrexone (LDN) on the epithelial mesenchymal transition of cervical cancer cells in vitro as well as its influence on macrophage polarization and associated cytokines in vivo. The results suggested that LDN supressed the proliferation, migration and invasion abilities and promote their apoptosis in Hela cells, whereas the opioid growth factor receptor (OGFr) silenced significantly reversed these effects in vitro. Knockdown the expression of OGFr, the inhibitory of LDN on EMT was weakened. LDN could inhibit cervical cancer progression in nude mice. In additon, LDN indirectly reduced the number of tumor-associated macrophages (TAMs), mainly M2 macrophages, and decreased expression of anti-inflammatory factor IL-10 in the serum of nude mice. These findings demonstrate that LDN could be a potential treatment for cervical cancer.     

黄芪桂枝五物汤对神经病理性疼痛大鼠脊髓CX3CR1表达的影响

目的研究黄芪桂枝五物汤对神经病理性疼痛大鼠的镇痛作用及对脊髓背角和背根节中脊髓小胶质细胞趋化因子受体1(CX3CR1)表达的影响。方法将50只SD大鼠随机分为模型组、黄芪桂枝五物汤高剂量组、黄芪桂枝五物汤中剂量组、黄芪桂枝五物汤低剂量组、塞来昔布组,每组10只。所有大鼠均建立坐骨神经慢性缩窄性损伤神经病理性疼痛模型,术后模型组给予生理盐水10 mL/kg灌胃,黄芪桂枝五物汤高、中、低剂量组分别给予黄芪桂枝五物汤0.2 mL/kg、0.1 mL/kg、0.05 mL/kg灌胃,塞来昔布组给予塞来昔布30 mg/kg灌胃,均1次/d,连续14 d。分别在造模前及术后3,5,7,14 d采用热刺痛仪测定各组大鼠的热缩足反射潜伏期,用免疫组化法检测各组大鼠脊髓背角和背根节中CX3CR1表达情况,用荧光定量PCR法和Western blot法分别检测脊髓背角和背根节中CX3CR1 mRNA和蛋白的相对表达量。结果术后3 d各组大鼠热缩足反射潜伏期均较术前1 d明显缩短(P均<0.05),但模型组和塞来昔布组术后各时间段比较差异无统计学意义(P均>0.05);黄芪桂枝五物汤低、中剂量组从术后7 d开始、黄芪桂枝五物汤高剂量组从术后3 d开始热缩足反射潜伏期均明显长于模型组(P均<0.05)。免疫组化显示黄芪桂枝五物汤各剂量组脊髓背角和背根节中CX3CR1阳性细胞均显著少于模型组和塞来昔布组(P均<0.05),且黄芪桂枝五物汤高剂量组均明显少于黄芪桂枝五物汤低、中剂量组(P均<0.05),黄芪桂枝五物汤低、中剂量组间比较差异无统计学意义(P均>0.05)。模型组和塞来昔布组脊髓背角和背根节中CX3CR1 mRNA和蛋白相对表达量比较差异无统计学意义(P均>0.05);黄芪桂枝五物汤各剂量组脊髓背角和背根节中CX3CR1 mRNA和蛋白相对表达量均显著低于模型组和塞来昔布组(P均<0.05),且黄芪桂枝五物汤高剂量组均明显低于黄芪桂枝五物汤低、中剂量组(P均<0.05),黄芪桂枝五物汤低、中剂量组间比较差异无统计学意义(P均>0.05)。结论黄芪桂枝五物汤对神经病理性疼痛模型大鼠具有显著镇痛作用,其可能通过调控脊髓背角和背根节CX3CR1的表达发挥作用,且呈一定剂量依赖性。