BACKGROUND: Psoriatic keratinocytes express CXC chemokines like IL-8 and GRO-alpha, and CC chemokines like MCP-1 and RANTES, which have a significant role in the accumulation of inflammatory cells in psoriatic skin and both CXCR1 and CXCR2 receptors are also expressed in psoriatic keratinocytes, which suggests that IL-8 and GRO-alpha could have a role in the characteristic epidermal changes through binding to their receptors in psoriatic keratinocytes. OBJECTIVE: The purpose is to understand the pathogenetic mechanisms of psoriasis by comparing immunoreactivity of various chemokines and chemokine receptors between lesional and non-lesional skin of psoriasis. METHODS:We have performed immunohistochemical studies with mouse anti-human IL-8, mouse anti-human GRO, anti-huamn MCP-1, mouse anti-human RANTES, anti-human CDw 128 IL-8RA/ CXCR1, and anti-human IL-8RB/CXCR2 for lesional and non-lesional skin of ten psoriatic patients. RESULTS: 1.Immunohistochemical reactivity for IL-8 is stronger in lesional epidermis than non-lesional epidermis(p<0.05) and immunohistochemical reactivity for GRO-alpha is stronger in lesional epidermis than non-lesional epidermis(p<0.05). 2.Immunohistochemical reactivity for MCP-1 is stronger in lesional epidermis than non-lesional epidermis(p<0.05), and immunohistochemical reactivity for RANTES is stronger in lesional epidermis than non-lesional epidermis(p<0.05). 3.Immunohistochemical reactivity for CXCR1 is stronger in lesional epidermis than non-lesional epidermis(p<0.05) and immunohistochemical reactivity for CXCR2 is stronger in lesional epidermis than non-lesional epidermis(p<0.05). 4.Immunofluorescent staining reveals positive finding in epidermis of lesional psoriasis, but negative finding in CXCR2. CONCLUSION: These results suggest that psoriatic keratinocytes express CXC chemokines like IL-8 and GRO-alpha, and CC chemokines like MCP-1 and RANTES, which have a significant role in the accumulation of inflammatory cells in psoriatic skin and that both CXCR1 and CXCR2 receptors are also expressed in psoriatic keratinocytes, which suggests that IL-8 and GRO-alpha could have a role in the characteristic epidermal changes through binding to their receptors in psoriatic keratinocytes.
Animals ; Chemokine CCL5 ; Chemokines* ; Chemokines, CC ; Chemokines, CXC ; Epidermis ; Humans ; Interleukin-8 ; Keratinocytes ; Mice ; Psoriasis* ; Receptors, Chemokine* ; Receptors, Interleukin-8B ; Skin

BACKGROUND: Psoriatic keratinocytes express CXC chemokines like IL-8 and GRO-alpha, and CC chemokines like MCP-1 and RANTES, which have a significant role in the accumulation of inflammatory cells in psoriatic skin and both CXCR1 and CXCR2 receptors are also expressed in psoriatic keratinocytes, which suggests that IL-8 and GRO-alpha could have a role in the characteristic epidermal changes through binding to their receptors in psoriatic keratinocytes. OBJECTIVE: The purpose is to understand the pathogenetic mechanisms of psoriasis by comparing immunoreactivity of various chemokines and chemokine receptors between lesional and non-lesional skin of psoriasis. METHODS:We have performed immunohistochemical studies with mouse anti-human IL-8, mouse anti-human GRO, anti-huamn MCP-1, mouse anti-human RANTES, anti-human CDw 128 IL-8RA/ CXCR1, and anti-human IL-8RB/CXCR2 for lesional and non-lesional skin of ten psoriatic patients. RESULTS: 1.Immunohistochemical reactivity for IL-8 is stronger in lesional epidermis than non-lesional epidermis(p<0.05) and immunohistochemical reactivity for GRO-alpha is stronger in lesional epidermis than non-lesional epidermis(p<0.05). 2.Immunohistochemical reactivity for MCP-1 is stronger in lesional epidermis than non-lesional epidermis(p<0.05), and immunohistochemical reactivity for RANTES is stronger in lesional epidermis than non-lesional epidermis(p<0.05). 3.Immunohistochemical reactivity for CXCR1 is stronger in lesional epidermis than non-lesional epidermis(p<0.05) and immunohistochemical reactivity for CXCR2 is stronger in lesional epidermis than non-lesional epidermis(p<0.05). 4.Immunofluorescent staining reveals positive finding in epidermis of lesional psoriasis, but negative finding in CXCR2. CONCLUSION: These results suggest that psoriatic keratinocytes express CXC chemokines like IL-8 and GRO-alpha, and CC chemokines like MCP-1 and RANTES, which have a significant role in the accumulation of inflammatory cells in psoriatic skin and that both CXCR1 and CXCR2 receptors are also expressed in psoriatic keratinocytes, which suggests that IL-8 and GRO-alpha could have a role in the characteristic epidermal changes through binding to their receptors in psoriatic keratinocytes.
Animals ; Chemokine CCL5 ; Chemokines* ; Chemokines, CC ; Chemokines, CXC ; Epidermis ; Humans ; Interleukin-8 ; Keratinocytes ; Mice ; Psoriasis* ; Receptors, Chemokine* ; Receptors, Interleukin-8B ; Skin

OBJECTIVETo explore the effects of trichloroethylene (TCE) and its by-products (trichloroacetic acid, TCA; dichloroacetic acid, DCA) on the normal human peripheral blood lymphocyte and the role of DCA in dermatitis medicamentosa- like induced by trichloroethylene (DMLT).

METHODSLymphocyte was isolated from peripheral venous blood, and cytotoxicity of human lymphocytes treated with different concentrations (0.02 approximately 30.00 mmol/L) of DCA was determined at indicated times (2 h and 4 h) based on the MTS assay. Action of DCA on cell viability, membrane integrity was assessed by neutral red uptake (NRU) assay and lactate dehydrogenase (LDH) release test and measurement of superoxide dismutase (SOD) activity. Fluorescence quantitative reverse transcription polymerase chain reaction (FQ-RT-PCR) was employed for detection and quantization of the chemokine receptor CXCR2 and chemokine receptor CXCR3 mRNA in peripheral blood lymphocyte treated with different concentrations of DCA.

RESULTSDCA had a more vital effect on peripheral blood lymphocyte than TCE and TCA. A concentration-dependent release of LDH was observed at 4 h after cells were exposed to different doses of DCA (0.88, 1.75, 3.50 and 7.00 mmol/L) (P < 0.05), and DCA also caused an inhibition of SOD activity in a concentration-dependent manner (P < 0.05). The results of FQ- RT- PCR indicated that CXCR2 and CXCR3 mRNA were all over- expression. At 48 h after the DCA of 0.5 mmol/L and 10.00 mmol/L was used, CXCR2 and CXCR3 mRNA were 10.34, 5.66-fold and 19.43, 8.75-fold of those in the control group (P < 0.01).

CONCLUSIONDCA is of a great cytotoxicity and may be one of crucial evocators on DMLT.

Adolescent ; Cells, Cultured ; Dichloroacetic Acid ; toxicity ; Female ; Humans ; Lymphocytes ; drug effects ; metabolism ; Male ; Receptors, CXCR3 ; metabolism ; Receptors, Interleukin-8B ; metabolism ; Trichloroethylene ; toxicity ; Young Adult

AIMTo validate the abundance of Interleukin 8 receptor beta (IL-8Rbeta) mRNA in single human neutrophil.

METHODSHuman neutrophils were isolated and purified from volunteers, total RNA was extracted and a regular RT-PCR aiming at IL-8Rbeta mRNA was performed to ascertain its expression profile in human neutrophils and optimize the reaction conditions for the following single-cell RT-PCR procedures. Subsequently, single neutrophil or the cellular content was harvested to conduct reverse transcription and two-round PCR with the same primer pairs used before. Serial dilution of single neutrophil cDNA pool was carried out at the same time with the exact two-round PCR followed. The specificity of this single-cell RT-PCR procedure was verified by the BamHI restriction endonuclease digestion on the final cDNA products.

RESULTSRegular RT-PCR indicated IL-8Rbeta mRNA expression in human neutrophils. While single-cell RT-PCR was sensitive enough to detect trace IL-8Rbeta mRNA as predicted cDNA product could be amplified from a 10 000 times diluted intracellular specimen from single neutrophil, which indicated an abundant expression of this mRNA in human neutrophil. Moreover, BamHI digestion on the final cDNA product clarified the specificity of this single-cell RT-PCR procedure.

CONCLUSIONThis simplified semi-quantitative single-cell RT-PCR procedure specifically confirmed that IL-8Rbeta mRNA was highly expressed in human neutrophil, which also provided the possibility of comparing mRNA abundance at single cell level.

Cells, Cultured ; Humans ; Neutrophils ; chemistry ; RNA, Messenger ; genetics ; Receptors, Interleukin-8B ; analysis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Single-Cell Analysis ; methods

This study was conducted to elucidate the effects of different fluid shear stress on the expression of IL-8 receptors CXCR1 and CXCR2 mRNA in endothelial cells, EA. Hy926 cells. The HUVEC cell lines were cultured in vitro and then exposed to 5.56, 10.02, and 15.27 dyn/cm2 fluid shear stress respectively for 5 min, 10 min, 15 min, 20 min, 25 min, 30 min, 1 h, 2 h, 4 h and 8 h. Semi-quantitative reversal transcription-polymerase chain reaction (RT-PCR) was used for detecting IL-8 receptors mRNA expression at different times. The results showed that, under 5. 56 dyn/cm2 shear stress, both the expression of CXCR1 and CXCR2 mRNA increased significantly with time (P<0.05). When exposed to 10. 02 dyn/cm2, the expression of CXCR1 mRNA was down-regulated with time on every occasion. CXCR2 mRNA increased temporally at 30 min, then it decreased gradually with time and finally went on at a constant lower level. When exposed to 15.27 dyn/cm2, both CXCR1 and CXCR2 mRNA expression decreased significantly with time (P<0.01). These data indicate that the expression of CXCR1 and CXCR2 mRNA of endothelial cell is regulated by fluid shear stress.
Cell Line ; Endothelial Cells ; cytology ; metabolism ; Gene Expression Regulation ; Humans ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Interleukin-8A ; biosynthesis ; genetics ; Receptors, Interleukin-8B ; biosynthesis ; genetics ; Shear Strength ; Stress, Mechanical ; Umbilical Veins ; cytology

CXCR1 and CXCR2 are important receptors in regulating vascular endothelial cell activities. In order to elucidate the role of CXCR1/2 in shear stress-induced endothelial cell migration, we have investigated the expression levels of CXCR1 and CXCR2 in the endothelial cells exposed to shear stress. In the experiment, anti-IL8RA and anti-IL8RB were used to antagonize CXCR1 and CXCR2. Different shear stresses were generated in a flow chamber; scratch test was carried out to compare endothelial cell migration in the control group and the receptor-antagonized groups. The results indicated that the migration of endothelial cells was restrained effectively after CXCR1 and CXCR2 were antagonized by anti-IL8RA and anti-IL8RB. And anti-IL8RA showed a stronger inhibitive effect than did anti-IL8RB (P<0.05). In the group with both receptor antagonisms, the migration was further inhibited. These results suggest that both CXCR1 and CXCR2 are important factors in mediating the migration of endothelial cells induced by shear stress, and CXCR1 fulfills a more important role.
Cell Movement ; physiology ; Endothelial Cells ; cytology ; metabolism ; Humans ; Mechanotransduction, Cellular ; drug effects ; Receptors, Interleukin-8A ; antagonists & inhibitors ; physiology ; Receptors, Interleukin-8B ; antagonists & inhibitors ; physiology ; Shear Strength ; Stress, Mechanical ; Umbilical Veins ; cytology ; metabolism

To observe the effect of acupuncture on CXCL8 receptors (CXCR1 and CXCR2) in rat endometrium experiencing embryo implantation failure, 72 pregnant rats were randomly divided into four groups: normal group (N), embryo implantation failure group (M), acupuncture treatment group (A), and progestin treatment group (W). Then the rats in each group were equally randomized into a day-6 (D6) group, a day-8 (D8) group, and a day-10 (D10) group. The rats in group M, group A, and group W were treated with mifepristone-sesame oil solution on day 1, while the rats in group N were injected with the same amount of sesame oil. Meanwhile, "Housanli" and "Sanyinjiao" were selected for acupuncture. From day 1 to the time of death, the rats in group A were fastened up and then acupuncture was administered while the rats in group N and group M were only fixed, and the rats in group W were given progestin. The number of implanted embryos was calculated. The expression of CXCR1 and CXCR2 in rat endometrium was detected by immunohistochemistry, Western blotting and real-time PCR. Compared to group N, the average number of implanted embryos, the protein and mRNA expression of CXCR1 (D6, D8 and D10), and the protein and mRNA expression of CXCR2 (D8 and D10) in rat endometrium were significantly decreased in group M. Compared to group M, there was significant elevation in the average number of implanted embryos, the protein expression (D6, D8 and D10) and mRNA expression (D8) of CXCR1 in rat endometrium of group A, and the protein expression (D8 and D10) and mRNA expression (D8) of CXCR2 in rat endometrium of group W. These findings indicated that acupuncture can increase the number of implanted embryos in rats of embryo implantation failure, which may be relevant with up-regulation the expression of CXCR1 and CXCR2 at maternal-fetal interface of rats with embryo implantation failure.
Acupuncture Therapy ; methods ; Animals ; Blotting, Western ; Embryo Implantation ; drug effects ; genetics ; Endometrium ; drug effects ; metabolism ; Female ; Gene Expression Regulation, Developmental ; Hormone Antagonists ; pharmacology ; Immunohistochemistry ; Mifepristone ; pharmacology ; Pregnancy ; Progestins ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar ; Receptors, Interleukin-8A ; genetics ; metabolism ; Receptors, Interleukin-8B ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors ; Treatment Outcome ; Up-Regulation ; drug effects