Neuropathic pain is one of the common complications of diabetes. Tetrahydropalmatine(THP) is a main active component of Corydalis Rhizoma with excellent anti-inflammatory and pain-alleviating properties. This study aims to investigate the therapeutic effect of THP on diabetic neuropathic pain(DNP) and the underlying mechanism. High-fat and high-sugar diet(4 weeks) and streptozotocin(STZ, 35 mg·kg~(-1), single intraperitoneal injection) were employed to induce type-2 DNP in rats. Moreover, lipopolysaccharide(LPS) was used to induce the activation of BV2 microglia in vitro to establish an inflammatory cellular model. Fasting blood glucose(FBG) was measured by a blood glucose meter. Mechanical withdrawal threshold(MWT) was assessed with von Frey filaments, and thermal withdrawal latency(TWL) with hot plate apparatus. The protein expression levels of OX42, inducible nitric oxide synthase(iNOS), CD206, p38, and p-p38 were determined by Western blot, the fluorescence expression levels of OX42 and p-p38 in the dorsal horn of the rat spinal cord by immunofluorescence, the mRNA content of p38 and OX42 in rat spinal cord tissue by qRT-PCR, and levels of nitric oxide(NO), interleukin-1β(IL-1β), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), interleukin-10(IL-10), and serum fasting insulin(FINS) by enzyme-linked immunosorbent assay(ELISA). RESULTS:: showed that the mo-del group demonstrated significant decrease in MWT and TWL, with pain symptoms. THP significantly improved the MWT and TWL of DNP rats, inhibited the activation of microglia and p38 MAPK signaling pathway in rat spinal cord, and ameliorated its inflammatory response. Meanwhile, THP promoted the change of LPS-induced BV2 microglia from the pro-inflammatory M1 phenotype to the anti-inflammatory M2 phenotype, suppressed the activation of the p38 MAPK signaling pathway, decreased the expression levels of inflammatory factors NO, IL-1β, IL-6, and TNF-α, and increased the expression level of anti-inflammatory factor IL-10. The findings suggested that THP can significantly ameliorate the pain symptoms of DNP rats possibly by inhibiting the inflammatory response caused by M1 polarization of microglia via the p38 MAPK pathway.
Animals ; Berberine Alkaloids ; Blood Glucose/metabolism* ; Diabetes Mellitus ; Diabetic Neuropathies/genetics* ; Interleukin-10 ; Interleukin-6/metabolism* ; Lipopolysaccharides/pharmacology* ; Microglia ; Neuralgia/metabolism* ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Spinal Cord/metabolism* ; Streptozocin/therapeutic use* ; Tumor Necrosis Factor-alpha/metabolism* ; p38 Mitogen-Activated Protein Kinases/metabolism*

PURPOSE: High mobility group box 1 (HMGB1) plays a central role in the pathogenesis of sepsis and multiple organ dysfunction syndromes. We investigated the associations of a single nucleotide polymorphism (SNP; rs1045411) in HMGB1 with various clinical parameters, severity, and prognosis in patients with sepsis, severe sepsis, or septic shock. MATERIALS AND METHODS: We enrolled 212 adult patients followed for 28 days. All patients were genotyped for rs1045411, and the serum levels of HMGB1 and several cytokines were measured. RESULTS: The proportions of patients according to genotype were GG (71.2%), GA (26.4%), and AA (2.4%). Among patients with chronic lung disease comorbidity, patients with a variant A allele had higher positive blood culture rates and higher levels of various cytokines [interleukin (IL)-1beta, IL-6, IL-10, IL-17, and tumor necrosis factor-alpha] than those with the GG genotype. In the analysis of those with diabetes as a comorbidity, patients with a variant A allele had higher blood culture and Gram-negative culture rates than those with GG genotypes; these patients also had a higher levels of IL-17. In the analysis of those with sepsis caused by a respiratory tract infection, patients with a variant A allele had higher levels of IL-10 and IL-17 (all p<0.05). This polymorphism had no significant impact on patient survival. CONCLUSION: The variant A allele of rs1045411 appears to be associated with a more severe inflammatory response than the GG genotype under specific conditions.
Adult ; Aged ; Alleles ; Asian Continental Ancestry Group/genetics ; China/epidemiology ; Cytokines/*blood/*genetics ; Female ; Genotype ; HMGB1 Protein/blood/*genetics ; Humans ; Interleukin-10/genetics ; Interleukin-17/genetics ; Interleukin-6/blood ; Male ; Middle Aged ; Polymorphism, Genetic/*genetics ; Polymorphism, Single Nucleotide/*genetics ; Prognosis ; Republic of Korea ; Sepsis/immunology/*metabolism/mortality ; Shock, Septic/immunology/*metabolism/mortality ; Survival ; Tumor Necrosis Factor-alpha/genetics

OBJECTIVETo study the infiltration of macrophages and their phenotype in the healing process of full-thickness wound in rat.

METHODSThirty healthy SD rats were divided into control group (n = 6) and injury group (n = 24) according to the random number table. Two round full-thickness skin defects (11 mm diameter) were created on both sides of dorsal spine of rats in injury group with surgical scissors and homemade trephine. After injury, wound area was measured immediately. The wounds were disinfected with iodophor every day. Rats in control group received anesthesia and hair removal only. On post injury day (PID) 1, 3, 7, and 13, respectively, 6 rats of injury group were sacrificed after the measurement of wound area (wound healing rate was calculated). Wound samples were obtained by excision down to healthy fascia along wound edge. Histological study was done with HE staining. The expression of CD68 (the surface marker of macrophage) in the wound tissue was observed with immunohistochemical staining. The double positive expressions of induced nitric oxide synthase (iNOS) plus CD68 (type I macrophage) and arginase 1 (Arg-1) plus CD68 (type II macrophage) were observed with immunofluorescence staining. The levels of interferon-γ (IFN-γ), TNF-α, IL-4, IL-13, IL-10, and IL-12 in wound tissue were assayed by double-antibody sandwich ELISA, and the ratio of IL-10/IL-12 was calculated. Full-thickness skin tissues (11 mm diameter) in rats of control group were excised at the same site as rats in injury group, and the histological observation and cytokines assay were performed as well. Data were processed with one-way analysis of variance or LSD- t test.

RESULTSWound area of rats in injury group was gradually reduced after injury, and the overall difference of the wound healing rate on each PID was statistically significant (F = 358.55, P < 0.01). No abnormal appearance of skin tissue was observed in rats of control group. In injury group, inflammatory cell infiltration was obvious in wound tissue on PID 1 and 3; vascular structure and fresh collagen were observed in wound tissue on PID 7 and 13. Numbers of CD68 positive cells in skin tissue of rats in control group and wound tissue of rats in injury group on PID 1, 3, 7, and 13 were respectively (2.7 ± 1.5), (31.8 ± 3.5), (40.8 ± 4.7), (20.8 ± 2.8), (3.2 ± 2.4) per 200 times visual field (F = 180.55, P < 0.01). Compared with that in control group, the number of CD68 positive cells of rats in injury group was increased on PID 1, 3, and 7 (with t values respectively 18.81, 18.79, 14.05, P values below 0.01). No double positive expression of iNOS plus CD68 or Arg-1 plus CD68 was observed in normal tissue of rats in control group. In injury group, proportions of iNOS plus CD68 double positive cells on PID 1, 3, 7, and 13 were respectively (12.2 ± 2.8)%, (16.5 ± 2.9)%, (4.2 ± 2.3)%, (0.7 ± 0.8)% (F = 72.50, P < 0.01); proportions of Arg-1 plus CD68 double positive cells on PID 1, 3, 7, and 13 were respectively 0, (8.2 ± 1.9)%, (21.5 ± 3.4)%, (4.7 ± 2.0)% (F = 120.93, P < 0.01). In injury group, proportion of iNOS plus CD68 double positive cells on PID 3 was significantly higher than that on other PID (with t values respectively 2.65, 8.17, 12.95, P values below 0.05); proportion of Arg-1 plus CD68 double positive cells on PID 7 was higher than that on other PID (with t values respectively 15.27, 8.25, 10.38, P values below 0.01). Compared with that of Arg-1 plus CD68 double positive cells, proportion of iNOS plus CD68 double positive cells was higher on PID 1 and 3 (with t values respectively 10.71 and 5.88, P values below 0.01) and lower on PID 7 and 13 (with t values respectively 10.24 and 4.60, P values below 0.01). The overall differences of IFN-γ, TNF-α, IL-4, IL-13, and IL-10/IL-12 ratio in skin tissue of rats in control group and wound tissue of rats in injury group on every PID were statistically significant (with F values from 14.08 to 631.03, P values below 0.01). Compared with those in control group, levels of IFN-γ, TNF-α, IL-4, and IL-13 in wound tissue of rats in injury group were significantly higher on every PID (with t values from 4.58 to 9.17, P values below 0.05), while IL-10/IL-12 ratio was significantly higher on PID 1, 3, and 7 (with t values respectively 27.70, 30.51, 9.49, P values below 0.05) . In injury group, IFN-γ level on PID 1 [(61 ± 5) pg/mL] and IL-10/IL-12 ratio on PID 3 (1.647 ± 0.098) were significantly higher than those of control group and those on other PID in injury group [with IFN-γ level respectively (32 ± 4), (54 ± 6), (46 ± 7), (47 ± 4) pg/mL and IL-10/IL-12 ratio respectively 0.328 ± 0.045, 0.960 ± 0.034, 0.530 ± 0.028, 0.289 ± 0.040, with t values respectively from 3.19 to 8.20 and from 16.59 to 31.84, P values below 0.05].

CONCLUSIONSMacrophage infiltration increases in the healing process of full-thickness wound in rat with different phenotypes, among which type I macrophage appears in the inflammatory stage, and type II macrophage predominates in the proliferative stage.

Animals ; Antigens, CD ; genetics ; metabolism ; Antigens, Differentiation, Myelomonocytic ; genetics ; metabolism ; Collagen ; Enzyme-Linked Immunosorbent Assay ; Interferon-gamma ; Interleukin-10 ; Interleukin-12 ; Interleukin-13 ; Interleukin-4 ; Macrophages ; Male ; Phenotype ; Rats ; Skin ; injuries ; Tumor Necrosis Factor-alpha ; blood ; Wound Healing ; genetics

OBJECTIVETo investigate effect of Chaiqin Chengqi Decoction (, CQCQD) on changes of neuronal acetylcholine receptor alpha 7 (nAChRα7) of peritoneal macrophages in acute pancreatitis (AP).

METHODSEighteen Kunming mice were equally randomized into the control group, AP group and CQCQD treatment group. AP was induced by two intraperitoneal injections of 4 g/kg L-arginine at 1 h apart, while control mice received saline injections. At 72 h after the first injection of L-arginine, mice in the treatment group were intragastrically administered 0.1 mL/10 g CQCQD every 2 h for 3 times, whilst mice in the other two groups received the same amount of saline feeding. Mice were sacrificed by cervical dislocation 2 h after the last feeding of either CQCQD or saline. Peritoneal macrophages were collected for determination of nAChRα7 mRNA and protein expression. Serum was collected for detection of interleukin-6 (IL-6), IL-10 and acetylcholine (ACh) levels, and pancreas was for histopathology analysis.

RESULTSThe CQCQD treatment significantly ameliorated the severity of AP as evidenced by reducing the pancreatic histopathology score (4.5±0.5 vs. 6.2±1.7, P<0.05) and the serum IL-6 levels (1228.3±419.2 pg/mL vs. 1589.6±337.3 pg/mL, P<0.05). The mRNA and protein expression of nAChRα7 of the peritoneal macrophages in the AP group were similar to the control group (P>0.05), but were significantly up-regulated after the CQCQD treatment (P<0.05). The serum ACh levels in the AP group were significantly lower than those in the control group (3.1±0.6 μg/mL vs 4.8±0.7 μg/mL P<0.05), but were significantly increased after the CQCQD treatment (5.6±1.5 μg/mL vs 3.1±0.6 μg/mL, P<0.05).

CONCLUSIONCQCQD is protective against L-arginine-induced AP through mechanisms involving nAChRα7 of peritoneal macrophages.

Acetylcholine ; pharmacology ; Acute Disease ; Animals ; Blotting, Western ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Interleukin-10 ; blood ; Interleukin-6 ; blood ; Macrophages, Peritoneal ; drug effects ; metabolism ; pathology ; Mice ; Neurons ; drug effects ; metabolism ; Pancreas ; drug effects ; pathology ; Pancreatitis ; blood ; drug therapy ; pathology ; RNA, Messenger ; genetics ; metabolism ; alpha7 Nicotinic Acetylcholine Receptor ; genetics ; metabolism

OBJECTIVE: To detect the expression of NGF, BDNF, NT-3 mRNA in the peripheral blood of patients with allergic rhinitis (AR). And to analyze the correlation between NGF, BDNF, NT-3 mRNA expression and the epidsode of rhinitis through Th-2 Hypothesis. METHOD: This study was a group controlled trial. The expression of NGF, BDNF and NT-3 mRNA were tested by real-time quantitative RT-PCR and the concentrations of IL-4, IL-6, IL-10 and INF-alpha were tested by ELISA. RESULT: The expression of NGF, BDNF and NT-3 mRNA in AR patients were 2.44, 4.46 and 1.78 times the amount of those in the healthy adults, respectively. The increased expression of NT-3 correlated positively with the scores of visual analog scale of AR. The concentrations of IL-4, IL-6 and IL-10, which were 2198 +/- 472 pg/mL, 9407 +/- 703 pg/mL and 3917 +/- 323 pg/mL respectively, were higher than those in the healthy adults. The concentration of INF-alpha was 2198 +/- 472 pg/mL and less than the healthy adults. The increased expressions of NGF, NT-3 were positively related to the increase of IL-4, IL-6 and IL-10. CONCLUSION The expressions of NGF, BDNF and NT-3 mRNA in AR patients are higher than those in the healthy adults. NGF, BDNF and NT-3 may contribute to the pathogenesis of AR. Moreover, NGF and NT-3 may induce the episode of rhinitis through Th-2 Hypothesis.
Adolescent ; Adult ; Brain-Derived Neurotrophic Factor ; blood ; Case-Control Studies ; Child ; Female ; Humans ; Interleukin-10 ; blood ; Interleukin-4 ; blood ; Interleukin-6 ; blood ; Male ; Nerve Growth Factor ; blood ; Nerve Growth Factors ; blood ; genetics ; Neurotrophin 3 ; blood ; RNA, Messenger ; genetics ; Rhinitis, Allergic ; blood ; immunology ; Th1-Th2 Balance ; Young Adult

Chronic inflammation is thought to be the leading cause of colorectal cancer, and interleukin-10 (IL10) has been identified as a potent immunomodulatory cytokine that regulates inflammatory responses in the gastrointestinal tract. Although several single nucleotide polymorphisms (SNPs) in IL10 have been associated with the risk of colorectal cancer, their prognostic significance has not been determined. Two hundred and eighty-two colorectal cancer patients were genotyped for two candidate cancer-associated SNPs in IL10. The associations of these SNPs with distant metastasis-free survival and overall survival were evaluated by Kaplan-Meier analysis and Cox regression model. The minor homozygote GG genotype of IL10 rs3021094 was significantly associated with a 3.30-fold higher risk of death compared with the TT+TG genotypes (P=0.011). The patients with IL10 rs3021094 GG genotype also had a poorer overall survival in Kaplan-Meier analysis (log-rank P=0.007) and in multivariate Cox regression model (P=0.044) adjusting for age, gender, carcinoembryonic antigen levels, tumor differentiation, stage, lymphovascular invasion, and perineural invasion. In conclusion, our results suggest that IL10 rs3021094 might be a valuable prognostic biomarker for colorectal cancer patients.
Aged ; Alleles ; Carcinoembryonic Antigen/blood ; Cell Differentiation ; Colorectal Neoplasms/*genetics/mortality/pathology ; Female ; Genotype ; Homozygote ; Humans ; Interleukin-10/*genetics ; Kaplan-Meier Estimate ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; *Polymorphism, Single Nucleotide ; Regression Analysis ; Tumor Markers, Biological/genetics

OBJECTIVETo observe the effect of Huanglian Jiedu IJecoction (HJU) on systemic and vascular immune responses of high fat diet fed apoE deficient (apoE(-/-)) mice.

METHODSEight wild type C57BL6 mice were recruited as the wild type common food group. Totally 24 apoE(-/-) mice were randomly divided into the ApoE'common food group, the ApoE(-/-) hyperlipidemia group, and the ApoE(-/-) hyperlipidemia plus HJD group, 8 in each group. In the present study, the common food mice and high fat fed mice were fed with a chow diet or a high cholesterol diet for 4 weeks. HJD was given to mice in the ApoE(-/-) hyperlipidemia plus HJD group at the daily dose of 5 g/kg by gastrogavage, while equal volume of pure water was given to mice in the rest groups by gastrogavage. Four weeks later, the plasma levels of blood lipids, the ratio of peripheral blood mononuclear cells, and expressions of Toll-like receptor 4 (TLR-4) and CD36 on the monocytes were detected. The pathological changes and expressions of cytokines in local aorta were detected. The plasma cytokine levels in response to lipopolysaccharide (LPS) were analyzed. Results (1) Compared with the wild type common food group, TO, TG, and LDL-O significantly increased in the ApoE(-/-) common food group (P < 0. 05, P < 0.01). Compared with the ApoE(-/-) common food group, TC and LDL-C significantly increased in the hyperlipidemia group (P < 0. 05). There was no statistical difference in each index between the ApoE(-/-) hyperlipidemia group and the ApoE(-/-) hyperlipidemia plus HJD group (P > 0.05). (2) Compared with the wild type common food group, no obvious change of the ratio of peripheral blood mononuclear cells happened, the TLR4 expression level significantly increased in the ApoE'common food group (P < 0. 05). Compared with the ApoE common food group, the ratio of peripheral blood mononuclear cells and the TLR4 expression level significantly increased in the ApoE' hyperlipidemia group (P < 0.05). Compared with the ApoE(-/-) hyperlipidemia group, the ratio of peripheral blood mononuclear cells and the TLR4 expression level significantly decreased. Besides, the CD36 expression level also significantly decreased (P<0.05). (3) After stimulated by LPS for 3 h, compared with the wild type common food group, plasma TNF-ct and IL-b expressions significantly increased in the ApoE(-/-) common food group (P < 0.05). Compared with the ApoE(-/-) common food group, plasma expressions of IL-12, TNF-alpha, MCP-1, and IL-10 increased, but with no statistical difference in the ApoE(-/-) hyperlipidemia group (P > 0.05). After 4-week intervention of HJD, compared with the ApoE(-/-) hyperlipidemia group, the MCP-1 expression was significantly down-regulated, while the IL-10 expression significantly increased, showing statistical difference (P < 0.05). Compared with the wild type common food group, mRNA expression levels of IFN-gamma, MCP-1 , TNF-alpha, IL-10, and IL-1beta significantly increased (P < 0. 05, P < 0.01). Compared with the ApoE(-/-) common food group, not only mRNA expression levels of IFN-gamma, MCP-1, TNF-alpha, and IL-1beta, further significantly increased, but also IL-12, IL-10, and TGF-beta significantly increased (P < 0. 05, P < 0. 01). After 4-week intervention of HJD, compared with the ApoE(-/-) hyperlipidemia group, mRNA expression levels of MCP-1, TNF-alpha, IL-1beta, and IL-12 significantly decreased in the ApoE(-/-) hyperlipidemia plus HJD group (P < 0.05, P < 0.01).

CONCLUSIONSHigh fat diet induced systemic reaction and inflammatory reactions of local vessels. The local inflammatory response of vessels exceeded systemic inflammatory response. Intervention of HJD could attenuate inflammatory response, especially in local arteries. Meanwhile, it enhanced systemic anti-inflammatory reactions.

Animals ; Aorta ; pathology ; Apolipoproteins E ; genetics ; CD36 Antigens ; metabolism ; Chemokine CCL2 ; metabolism ; Dietary Fats ; adverse effects ; Drugs, Chinese Herbal ; pharmacology ; Female ; Hyperlipidemias ; blood ; etiology ; immunology ; Inflammation ; Interleukin-10 ; blood ; Interleukin-12 ; blood ; Interleukin-1beta ; blood ; Leukocytes, Mononuclear ; metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Systemic Inflammatory Response Syndrome ; blood ; etiology ; immunology ; Toll-Like Receptor 4 ; metabolism ; Transforming Growth Factor beta ; blood ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo study the immunoregulatory effect of hepatitis B virus (HBV) e antigen (HBeAg) on peripheral blood monocytes (PBMCs).

METHODSPBMCs were isolated from patients with chronic hepatitis B (CHB; both HBeAg- and HBeAg+) and healthy controls, and cultured with recombinant HBeAg. The HBeAg-induced changes in expression of PD-1/PD-L1 were measured by flow cytometry of the cells and in secreted cytokines were measured by enzyme-linked immunosorbent assay of the supernatants. Comparisons between two groups were made by the independent-samples t-test; the relationship between PD-1/B7-H1 level and HBV DNA copy number was evaluated by Spearman's correlation analysis.

RESULTSExposure to HBeAg led to a significant decrease in CD3+CD4+ T lymphocyte-specific expression of IFNa for both the CHB patients' and healthy controls' samples (t = 2.382 and -4.190 respectively, P less than 0.01). For the HBeAg- CHB patients' and healthy controls' samples, the HBeAg exposure led to increased levels of secreted cytokines IL-6, IL-10 and TNFa (t = 2.504, 3.583 and 4.324, P less than 0.01 and t = 3.542, 6.246 and 5.273, P less than 0.01 respectively) and of CD14+ PBMC-specific expression of PD-L1 (t = 4.815 and 3.454, P less than 0.05 respectively). Compared to the HBeAg-negative CHB patients' and healthy controls' samples, the HBeAg+ CHB patients' samples had significantly lower CD3+CD4+ T cell-specific expression of IFNa (t = -3.177 and -4.541, P less than 0.01 respectively), but significantly higher levels of secreted IL-4 (t = 3.382 and 4.393, P less than 0.01 respectively), of CD3+ T cells-specific expression of PD-1/PD-L1 (t = 4.755, 2.942 and 4.518, 4.595, P less than 0.01 respectively), and of CD14+ T cells-specific expression of PD-L1 (t = 5.092 and 5.473, P less than 0.01 respectively). The CD3+ T cells-specific expression of PD-L1 was significantly higher in the samples from HBeAg- CHB patients than from the healthy controls (t = 3.214, P less than 0.01).

CONCLUSIONHBeAg was able to down-regulate the production of Th1-type cytokines (IFNgamma), and up-regulate the secretion of Th2-type cytokines (IL-6, IL-10) and the expression of PD-1/PD-L1on monocytes. These changes are conducive to the formation of immune tolerance to HBV. Therefore, HBeAg may play an important role in immune tolerance to chronic HBV infection.

Adult ; Case-Control Studies ; Cells, Cultured ; Female ; Hepatitis B e Antigens ; genetics ; immunology ; Hepatitis B, Chronic ; blood ; immunology ; Humans ; Interferon-gamma ; immunology ; Interleukin-10 ; immunology ; Interleukin-6 ; immunology ; Leukocytes, Mononuclear ; immunology ; metabolism ; Male ; Middle Aged ; Recombinant Proteins ; immunology ; Th1 Cells ; immunology ; Th1-Th2 Balance ; Th2 Cells ; immunology

OBJECTIVETo explore the regulatory mechanism of Xiaoyin Recipe () on the T helper 1/T helper 2 (Th1/Th2) immune balance.

METHODSThirty-six experimental animals were divided into three groups, 12 rats in each group: blank control group (B group), negative control group (N group), and Xiaoyin Recipe treatment group (T group). The latter two groups received immunization of experimental autoimmune thyroiditis (EAT), and T group were treated with Xiaoyin Recipe for a month. Then, the expression of Th1-Th2-related genes in peripheral blood mononuclear cells (PBMCs) were screened with Oligo GEArray Rat Th1-Th2-Th3 Microarray. The expressions of tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), T-box expressed in T-cells (T-bet), and GATA-binding protein-3 (GATA-3) were detected by real-time polymerase chain reaction (RT-PCR).

RESULTSGene array screening showed that compared to N group, in T group after Xiaoyin Recipe treatment, 3 genes were upregulated in EAT rats, including interleukin-27 receptor alpha (IL-27rα), glomulin (Glmn), and GATA-3, while 38 genes were downregulated, such as CD28, IL-18, signal transducer, and activator of transcription 1 (STAT1), T-bet, TNF receptor superfamily member 4 (TNFRSF4), TNF ligand superfamily member 5 (TNFSF5), and TNF receptor superfamily member 5 (TNFRSF5). While RT-PCR showed that there was an increased level of TNF-α mRNA (P<0.01), an elevated ratio of T-bet/GATA-3, and a decreased level of IL-10 mRNA in PBMC of N and T group compared to B group (P <0.01); and after treatment with Xiaoyin Recipe, IL-10 mRNA level increased (P <0.01), while TNF-α mRNA level and T-bet/GATA-3 ratio decreased in T group compared to N group (P <0.01).

CONCLUSIONXiaoyin Recipe for psoriasis could induce a Th1/Th2 balance drift toward Th2 in PBMC of EAT rats and thus improve the conditions.

Animals ; Autoantibodies ; blood ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; GATA3 Transcription Factor ; genetics ; metabolism ; Gene Expression Regulation ; drug effects ; Interleukin-10 ; genetics ; metabolism ; Psoriasis ; blood ; drug therapy ; immunology ; pathology ; Rats ; Rats, Wistar ; T-Box Domain Proteins ; genetics ; metabolism ; Th1-Th2 Balance ; drug effects ; Th2 Cells ; drug effects ; immunology ; Thyroiditis, Autoimmune ; blood ; drug therapy ; immunology ; pathology ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

BACKGROUNDWhile the incidence of paroxysmal nocturnal hemoglobinuria (PNH) is relatively high in Northern China, the exact mechanism of the disease remains unknown. Immunoregulatory cytokine polymorphisms can directly regulate the expression levels of cytokines, which play a crucial role in many diseases. The purpose of this study was to study cytokine gene single nucleotide polymorphisms (SNPs) and the correlated cytokine expression levels in relationship to the PNH pathogenesis.

METHODSPeripheral blood samples were collected from 30 PNH patients and 40 healthy donors; all of the samples were collected from the Han people of Northern China. Eight SNP loci in five cytokine genes, including tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), transforming growth factor-beta (TGF-β), interleukin-6 (IL-6), and IL-10, and aplastic anemia (AA) were assessed. TNF-a, TGF-b, IFN-g, IL-6, and IL-10 were analyzed by sequence-specific primer polymerase chain reaction (PCR-SSP). The plasma protein levels of TNF-a, TGF-b, and IFN-g were assessed by an ELISA.

RESULTSThe PNH patients had a lower frequency of the TC/GG genotype of the TGF-b gene (P < 0.01) and a higher frequency of the C allele in the TGF-b gene (+10) compared to the controls (P < 0.05). The predominant genotype of the +874 locus of the IFN-g gene was TA in the PNH patients, while that in the predominant genotype was AA in the control group and was statistically significant (P < 0.001). The frequency of the T allele in the IFN-g gene was dramatically higher in the PNH patients than in the controls (P < 0.05). The PNH patients had a reduced frequency of the GC and CC genotypes, as well as the C allele at locus -174 of the IL-6 gene compared to the controls (P < 0.01). In addition, the plasma concentrations of TNF-a, TGF-b, and IFN-g were significantly higher in the PNH group compared to the control group (P < 0.01).

CONCLUSIONSExpression levels of the TNF-a, TGF-b, and IFN-g cytokines play an important role in PNH. The GC and CC genotypes, as well as the C allele of the IL-6 gene may protect the Han people of Northern China against PNH. Additionally, the TC/GG genotype of the TGF-b gene may be the protective allele. In contrast, the TA genotype and the T allele for the IFN-g gene, as well as the C allele of TGF-b may be susceptible to PNH. However, SNPs in the TNF-a and IL-10 genes did not correlate with PNH development. Alternatively, the increased plasma concentrations of TNF-a, TGF-b, and IFN-g in PNH patients may also be related to PNH development.

Adult ; Aged ; Alleles ; Anemia, Aplastic ; genetics ; Asian Continental Ancestry Group ; China ; Cytokines ; blood ; genetics ; Enzyme-Linked Immunosorbent Assay ; Female ; Genotype ; Hemoglobinuria, Paroxysmal ; blood ; genetics ; Humans ; Interferon-gamma ; blood ; genetics ; Interleukin-10 ; blood ; genetics ; Interleukin-6 ; blood ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic ; genetics ; Polymorphism, Single Nucleotide ; genetics ; Transforming Growth Factor beta ; genetics ; Tumor Necrosis Factor-alpha ; blood ; genetics ; Young Adult