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Liver fibrosis is a common reversible pathological change in chronic liver injury and may progress to liver cirrhosis, liver failure, and portal hypertension. In recent years, several studies have shown a significant change in chemokine profiles in patients with liver fibrosis, which is closely associated with the progression of liver fibrosis. Monocyte chemoattractant protein 1 (MCP-1) belongs to the family of CC chemokines and can induce the activation, recruitment, and migration of inflammatory cells during liver fibrosis. MCP-1 may be involved in the activation of hepatic stellate cells, the development of insulin resistance, and the progression to hepatocellular carcinoma. This article mainly reviews the potential role of MCP-1 and CC chemokine receptor in the progression of liver fibrosis and related therapies.
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Objective To evaluate the role of monocyte chemotactic factor-1 (MCP-1)∕chemokine C-C receptor 2 ( CCR2) in amygdala in neuropathic pain ( NP) in rats. Methods Clean-grade healthy male Sprague-Dawley rats, weighing 200-260 g, aged 2 months, in which NP model was established by ligating the left L5,6spinal nerve, were used in this study. The experiment was performed in two parts. Ex-periment Ⅰ Thirty-two rats were divided into 2 groups using a random number table method: control group (C group, n＝8) and NP group (n＝24). Rats were sacrificed at 7, 14 and 21 days after establis-hing NP model in group NP or at the corresponding time points before establishing NP model in group C, and the amygdala was removed for determination of the expression of MCP-1 and CCR2 mRNA and positive cell count using quantitative real-time polymerase chain reaction and immunohistochemistry. Experiment ⅡThirty-two rats were divided into 4 groups ( n＝8 each) using a random number table method: control group (C group), NP group, MCP-1 group and specific CCR2 antagonist RS102895 group (RS group). MCP-1 (50 pmol for each side) or RS102895 (100 pmol for each side) was injected into the bilateral a-mygdala at days 3, 6, 13 and 20 after establishing NP model in MCP-1 and RS groups, respectively. The thermal paw withdrawal latency (TWL) and mechanical paw withdrawal threshold (MWT) were measured at days 4, 7, 14 and 21 after establishing NP model (T1-4). Rats were sacrificed at T4and the L5segment of the spinal cord was removed for determination of interleukin-1beta (IL-1β), IL-6 and tumor necrosis fac-tor-alpha ( TNF-α) contents by enzyme-linked immunosorbent assay. Results Experiment Ⅰ Compared with group C, the expression of MCP-1 and CCR2 mRNA in amygdala was significantly up-regulated, and the number of MCP-1 and CCR2 positive cells was increased in group NP ( P＜0. 05). Experiment ⅡCompared with group C, the MWT was significantly decreased and TWL was shortened at T1-4, and the contents of IL-1β, IL-6 and TNF-α were increased in the other three groups ( P＜0. 05). Compared with group NP, the MWT was significantly decreased and TWL was shortened at T1, and the contents of IL-1β, IL-6 and TNF-α were increased in group MCP-1, and the MWT was significantly increased and TWL was prolonged at T1-4, and the contents of IL-1β, IL-6 and TNF-α were decreased in group RS ( P＜0. 05 or 0. 01). Conclusion Enhanced function of MCP-1∕CCR2 in amygdala may be involved in the pathophysio-logical process of NP in rats.
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Objective To develop and validate a ultrasonographic (US)imaging agent with targeted microbubbles that attaches to chemokine receptor 2 (CCR2)and to compare the US single obtained from targeted microbubble with that from control microbubble in murine breast tumor model.Methods The microbubble which carried CCR2 antibody (MBCCR2 )and isotype-macthed immunoglobulin G-labled control microbubble (MBcontrol ) were prepared. The microbubble size and distribution were assessed by AccuSizer780.Binding specificities of targeted microbubble compared with control microbubble were tested with murine microvascular endothelial cells (bEnd.3 ).Orthotopic breast tumor model was estabished in BALB/c mice with mouse breast cancer 4T1 cell.In vivo imaging signals of contrast material-enhanced ultrasound by use these two different types of microbubble which were injected respectively into each mouse at random order and 30 min interval.Tumor tissue was stained for CCR2 and CD3 1 .Results Automatic Particle Sizer showed size uniform of two kinds of microbubbles,and narrow distribution of particle size (mean diameter of about 1 -2 μm),which were not significantly different (P >0.05).Adhension to bEnd. 3 endothelial cells was significantly higher (P < 0.001 )for MBCCR2 (mean,9.50 ± 1 .5 1 )than that for MBcontrol (mean,0.01 ±0.01).Imaging signal in the murine tumor model was significantly higher for MBCCR2 [mean,(6.76±0.26)dB]than that for MBcontrol [mean,(1 .06 ±0.62)dB,P <0.001 ].Immunofluorescence confirmed expression of CCR2 on tumor vasculature.Conclusions The targeted microbubbles with CCR2 monoclonal antibody had been successfully prepared,which precisely targeted to CCR2 of tumor angiogenesis in the murine breast cancer xenograft tumor models in vivo.These results suggest that the targeted microbubbles as a kind of ultrasound molecular imaging agent with a better specificity can be used for both evaluating tumor neovascularization and monitoring therapeutic effect of anti-angiogenesis.
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Objective To evaluate the relationship between hippocampal monocyte chemotactic protein-1 (MCP-1) and its receptor C-C chemokine receptor 2 (CCR2) and postoperative cognitive dysfunction (POCD) in aged rats.Methods Forty-eight male Sprague-Dawley rats,aged 20-22 months,weighing 480-550 g,were randomly divided into 2 groups (n =24 each) using a random number table:control group (group C) and POCD group.POCD group inhaled 2.0％ isoflurane and underwent splenectomy.Before surgery and at 1,3 and 7 days after surgery,Morris water maze test was performed to evaluate the spatial learning and memory ability.The escape latency and swimming distance were recorded.Eight rats were sacrificed after the end of Morris water maze test performed at 1,3 and 7 days after surgery.Then the brains were removed,and the hippocampi were isolated for detection of the expression of MCP-1 and CCR2 by Western blot.Results Compared with group C,the escape latency and swimming distance were significantly prolonged,and the expression of MCP-1 and CCR2 in hippocampi was up-regulated at 1,3 and 7 days after surgery in POCD group.Conclusion Up-regulation of hippocampal MCP-1 and CCR2 expression may be involved in the mechanism of POCD in aged rats.
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Objective To explore the relationship between the expression of chemokines and their receptors in the maternal-fetal interface and the pathogenesis of unexplained recurrent spontaneous abortion (URSA). Methods 8-10 weeks CBA/J female mice were mated with DBA/2 and BALB/c male mice at the ratio of 2∶1 to establish the model of normal pregnant mice (CBA/J × BALB/c) and URSA mice (CBA/J × DBA/2). Sixty mice were divided into 6 groups, with ten in each group. The mice in the normal unpregnancy group were executed for endometrial tissues; the mice in the embryonic implantation normal pregnancy group were executed for endometrial tissues at the sixth day of gestation; the mice in the embryonic development normal pregnancy group were executed for decidua and chorionic tissues at the fourteenth day of gestation. While, the mice in the embryonic implantation URSA group were executed for endometrial tissues at the sixth day of gestation;the mice in the pre-abortion URSA group were executed for decidua and chorionic tissues at the ninth day of gestation;the mice in the post-abortion URSA group were executed for decidua and chorionic tissues at the fourteenth day of gestation. The chemokines and their receptors in different tissues of the mice were determined by western blot, including the protein expression of stromal cell derived factor (CXCL12), monocyte chemotactic protein 1 (CCL2), regulated upon activation normal T cell expressed and secreted(RANTES) and their receptor CXCR4, CCR2, CCR5 in maternal-fetal interface. Results (1) The protein expression of CXCL12 and CXCR4, CCL2 and CCR2, RANTES and CCR5 in endometrial tissues of the normal unpregnant group were 0.13±0.04 and 0.18±0.09, 0.057±0.023 and 0.39± 0.08, 0.034 ± 0.012 and 0.22 ± 0.05, respectively. They were 0.35 ± 0.09 and 0.93 ± 0.15, 0.349 ± 0.056 and 0.91 ± 0.15, 0.336 ± 0.089 and 0.44 ± 0.05 in endometrial tissues in the embryonic implantation normal pregnancy group;and were 0.62±0.15 and 1.23±0.28, 0.283±0.051 and 0.55±0.09, 0.225±0.065 and 0.35± 0.07 in decidua tissues in the embryonic development normal pregnancy group. The protein expression of chemokines and their receptors in endometrial tissues in the embryonic implantation normal pregnancy group and in decidua tissues in the embryonic development normal pregnancy group were higher than those in the normal unpregnancy group, with statistically significant difference(P<0.05). Compared with the embryonic implantation normal pregnancy group, CXCL12 and CXCR4 in decidual tissues in the embryonic development normal pregnancy group were significantly higher(P<0.05), while CCL2 and CCR2, RANTES and CCR5 were significantly lower (P<0.05). (2) Compared with the embryonic implantation normal pregnancy group, CXCL12 and CXCR4 (0.20±0.06 and 0.44±0.11) in endometrial tissues in the embryonic implantation URSA group were significantly lower (P<0.01), while CCL2 and CCR2(0.451±0.133 and 1.32± 0.20), RANTES and CCR5(0.488 ± 0.137 and 0.61 ± 0.18)were higher (P<0.05). (3) Compared with the embryonic development normal pregnancy group, CXCL12 and CXCR4 in decidual tissues of pre-abortion URSA group(0.27 ± 0.09 and 0.26 ± 0.10) , post-abortion URSA group (0.25 ± 0.08 and 0.23 ± 0.08) were significantly lower (P<0.01), while CCL2 and CCR2 (0.576±0.123 and 0.92±0.15 in the pre-abortion URSA group;0.748±0.112 and 1.56±0.34 in the post-abortion URSA group), RANTES and CCR5(0.294±0.054 and 0.59 ± 0.18 in the pre-abortion URSA group;0.363 ± 0.058 and 0.78 ± 0.14 in the post-abortion URSA group) were significantly higher(P<0.05). CCL2 and CCR2, RANTES and CCR5 in decidual tissues in the post-abortion URSA group was obviously higher than those of the pre-abortion URSA group, with statistically significant difference (P<0.05). Couclusions The accurate expression of CXCL12, CCL2, RANTES and their receptors CXCR4, CCR2, CCR5 play important roles in the embryonic implantation and development. The lower expression of CXCL12 and CXCR4 protein and higher expression of CCL2 and CCR2, RANTES and CCR5 in decidua and chorionic tissues are closely related to the pathogenesis of URSA.
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Objective To evaluate the effects of intrathecal TRESK gene recombinant adenovirus on inflammatory responses mediated by chemokine in the spinal cord of rats with neuropathic pain ( NP ) . Methods Thirty?six male Sprague?Dawley rats, weighing 200-250 g, were randomly divided into 6 groups (n=6 each) using a random number table: control group (group C); sham operation group (group S);NP group; TRESK?overexpressed adenovirus group ( group TRESK ); negative adenovirus group ( group Virus); normal saline group ( group NS) . Spinal nerve injury was produced by exposing the sciatic nerve and its branches and ligation and transection of tibial nerve and common fibular nerve in anesthetized rats. In TRESK, Virus and NS groups, pAd∕CMV∕V5?DEST?TRESK 25 μl (109IU∕ml), negative adenovirus 25 μl and normal saline 25 μl were intrathecally injected, respectively. At 1 day before operation ( base?line, T0 ) and 1, 3, 7 and 14 days after operation ( T1-4 ) , the mechanical paw withdrawal threshold ( MWT) and thermal paw withdrawal latency were measured. Six rats in each group were sacrificed after measurement of pain threshold at T3 . The L4,5 segments of the spinal cords were removed for determination of monocyte chemotactic protein?1 ( MCP?1) , MIP?2, tumor necrosis factor?alpha ( TNF?α) , interleukin?1 beta ( IL?1β) and IL?6 mRNA expression by real?time PCR. Results There was no significant difference in thermal paw withdrawal latency at each time point between groups. Compared with C and S groups, MWT at T1-4 in NP and TRESK groups and at T1-3 in Virus and NS groups were significantly decreased, and the expression of MCP?1, MIP?2, TNF?α, IL?1βand IL?6 mRNA was up?regulated in NP, TRESK, Virus and NS groups. Compared with group NP, MWT was significantly increased at T1-4, and the expres?sion of MCP?1, MIP?2, TNF?α, IL?1β and IL?6 mRNA was down?regulated in group TRESK. Conclusion The mechanism by which intrathecal TRESK gene recombinant adenovirus reduces NP is re?lated to inhibition of inflammatory responses mediated by chemokine in the spinal cord of rats.
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Objective To investigate the relationship between C-C chemokinereceptor type 2 (CCR2) and p38 mitogen-activated protein kinase (p38MAPK) signaling pathway in the dorsal root ganglion of rats and further clarify the mechanism of inflammatory pain.Methods Sevemy-two female Sprague-Dawley rats,weighing 150-180 g,aged 3-4 months,were randomly divided into 3 groups (n =24 each) using a random number table:control group (group C),inflammatory pain group (IP group) and CCR2 inhibitor RS102895 group (group RS).Inflammatory pain was induced by subcutaneous injection of Freund's adjuvant 100μl into the plantar surface of the right hindpaw.RS102895 20 mg/kg was injected subcutaneously once a day for 7 consecutive days in addition to Freund's adjuvant in group RS.Mechanical paw withdrawal threshold (MWT) was measured before injection and at 1,3,5 and 7 days after injection.At 3,5 and 7 days after injection,8 rats in each group were sacrificed and the dorsal root ganglions were removed for determination of the expression of CCR2 and phosphor-p38MAPK (p-p38MAPK) (by immuno-histochemical staining),and CCR2 and p38MAPK mRNA (using fluorescent quantitative PCR).Immuno-histochemical staining was scored.Results Compared with group C,MWT was significantly decreased after injection,the number of p-p38MAPK positive neurons and immuno-histochemical staining score were increased,and CCR2 and p38MAPK mRNA expression was up-regulated in IP and RS groups.Compared with group IP,MWT was significantly increased after injection,the number of p-p38MAPK positive neurons and immuno-histochemical staining score were decreased,and CCR2 and p3gMAPK mRNAexpression was down-regulated in RS group.Conclusion CCR2 in the dorsal root ganglion is involved in the development of inflammato pain possibility through activating p38MAPK signaling pathway in rats.
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Objective To evaluate the role of spinal microglial C-C chemokine receptor type 2 (CCR2) in the maintenance of bone cancer pain (BCP) in rats.Methods Fifty unmated female Sprague-Dawley rats,aged 2 months,weighing 160-180 g,were randomly divided into 5 groups (n =10 each):sham operation group (group Ⅰ),sham operation + RS102895 (CCR2 antagonist) group (group Ⅱ),BCP group (group Ⅲ),BCP + dimethylsulfoxide (DMSO) group (group Ⅳ),and BCP + RS102895 group (group Ⅴ).The rats were anesthetized with intraperitoneal chloral hydrate.BCP was induced by intra-tibial inoculation of 1 × 105 Walker 256 mammary gland carcinoma cells into the medullary cavity of the left tibial metaphysis.On 10-12 days after operation,3 μg/μl RS102895 10 μd was injected intrathecally once a day in Ⅱ and Ⅴ groups,10％ DMSO 10 μl was injected intrathecally once a day in Ⅳ group,and normal saline 10 μl was injected intrathecally once a day in Ⅰ and Ⅲ groups.Mechanical paw withdrawal threshold was measured at 1 day before operation and 3,6,9,10,11 and 12 days after operation.After measurement of pain threshold at day 12 after operation,the lumbar segments (L4-6) of the spinal cord were removed for determination of the level of ox-42 (spinal microglial activation marker) (by immuno-histochemistry) and contents of IL1-β,IL-6 and TNF-α (by ELISA).Results Compared with group Ⅰ,mechanical paw withdrawal threshold was significantly decreased at 6-12 days after operation,the number of ox-42 positive cells and contents of IL1-β,IL-6 and TNF-α were increased in Ⅲ,Ⅳ and Ⅴ groups,and no significant change was found in the parameters mentioned above in group Ⅱ.Compared with group Ⅲ,mechanical paw withdrawal threshold was significantly increased at 10-12 days after operation,the number of ox-42 positive cells and contents of IL1-β,IL-6 and TNF-α were decreased in group Ⅴ,and no significant change was found in the parameters mentioned above in group Ⅳ.Conclusion Spinal microglial CCR2 is involved in the maintenance of BCP via activating microglia and promoting the release of inflammatory cytokines of rats.
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Objective To investigate the effect of propofol pretreatment on hippocampal monocyte chemotactic protein-1 ( MCP-1 ) and CC-chemokine receptor type 2 (CCR2) expression following forebrain ischemiarepcrfusion (I/R) in rats. Methods Twenty-four male SD rats weighing 250-300 g were randomly divided into 3 groups ( n = 8 each): group Ⅰ control; group Ⅱ I/R and group Ⅲ propofol pretreatment. Cerebral I/R was induced by clamping bilateral common carotid arteries for 10 min combined with hypotension ( MAP was maintained at 35-45 mm Hg) induced by exsanguinations in group Ⅱ and Ⅲ. In group Ⅲ propofol 50 mg/kg was injected into femoral vein immediately before cerebral ischemia. The animals were sacrificed at 6 h of reperfusion. Hippocampal tissue was obtained for detection of MCP-1 mRNA and CCR2 mRNA and their protein expression by RT-PCR and Western blot technique. Results I/R significantly increased the expression of MCP-1 and CCR2 in hippoeampal tissue as compared with control group. Propofol pretreatment significantly attenuated cerebral I/R induced increase in MCP-1 and CCR2 expression. Conclusion Propofol pretreatment can significantly inhibit forebrain I/R-induced hippocampal MCP-1 and CCP2 expression.