Humans ; Inflammatory Bowel Diseases/drug therapy* ; Interleukin-6/therapeutic use*

PURPOSE: The combined use of antibiotics and anti-inflammatory medicine to manage bacterial endotoxin-induced inflammation following injuries or diseases is increasing. The cytokine level produced by macrophages plays an important role in this treatment course. Ciprofloxacin and indomethacin, two typical representatives of antibiotics and anti-inflammatory medicine, are cost-effective and has been reported to show satisfactory effect. The current study aims to investigate the effect of ciprofloxacin along with indomethacin on the secretion of inflammatory cytokines by macrophages in vitro. METHODS: Primary murine peritoneal macrophages and RAW 264.7 cells were administrated with lipopolysaccharide (LPS) for 24 h. The related optimal dose and time point of ciprofloxacin or indomethacin in response to macrophage inflammatory response inflammation were determined via macrophage secretion induced by LPS. Then, the effects of ciprofloxacin and indomethacin on the secretory functions and viability of various macrophages were determined by enzyme-linked immunosorbent assay and flow cytometry analysis, especially for the levels of interleukin (IL)-1β, IL-6, IL-10, and tumor necrosis factor (TNF)-α. The optimal dose and time course of ciprofloxacin affecting macrophage inflammatory response were determined by testing the maximum inhibitory effect of the drugs on pro-inflammatory factors at each concentration or time point. RESULTS: According to the levels of cytokines secreted by various macrophages (1.2 × 10⁶ cells/well) after administration of 1 μg/mL LPS, the optimal dose and usage timing for ciprofloxacin alone were 80 μg/mL and 24 h, respectively, and the optimal dose for indomethacin alone was 10 μg/mL. Compared with the LPS-stimulated group, the combination of ciprofloxacin and indomethacin reduced the levels of IL-1β (p < 0.05), IL-6 (p < 0.05), IL-10 (p < 0.01)), and TNF-α (p < 0.01). Furthermore, there was greater stability in the reduction of inflammatory factor levels in the combination group compared with those in which only ciprofloxacin or indomethacin was used. CONCLUSION The combination of ciprofloxacin and indomethacin suppressed the levels of inflammatory cytokines secreted by macrophages in vitro. This study illustrates the regulatory mechanism of drug combinations on innate immune cells that cause inflammatory reactions. In addition, it provides a new potential antibacterial and anti-inflammatory treatment pattern to prevent and cure various complications in the future.
Humans ; Mice ; Animals ; Cytokines ; Lipopolysaccharides/pharmacology* ; Interleukin-10 ; Indomethacin/therapeutic use* ; Interleukin-6/therapeutic use* ; Ciprofloxacin/therapeutic use* ; Macrophages ; Tumor Necrosis Factor-alpha ; Inflammation/drug therapy* ; Anti-Inflammatory Agents/therapeutic use* ; Anti-Bacterial Agents/therapeutic use*

BACKGROUNDChai Lai Prescription is a Chinese herbal compound which is used to sooth the liver, strengthen the spleen and harmonize the stomach for descending adverse Qi. We initiated the study to investigate its mechanism of treating in vitro rabbit reflux esophagitis models.

METHODSAdult male Japanese white rabbits, weighing 1.8-2.2 kg, were divided into five groups of three each, which were: normal control group (Krebs buffer, pH7.4), esophagitis model group (Krebs buffer, pH5.8), esophagitis model proup+low-dose Chinese herbal medicine protection group (0.6 mg × ml(-1)× kg(-1)), esophagitis model group+moderate-dose Chinese herbal medicine protection group (6 mg × ml(-1)× kg(-1)), esophagitis model group+high-dose Chinese herbal medicine protection group (60 mg × ml(-1)×kg(-1)). The RT-PCR method was used to test the influence of Chai Lai Prescription on IL-1 and IL-6 in in vitro rabbit models of esophagitis. We treated the in vitro models with different doses of Chinese herbal medicine.

RESULTSEsophageal mucosa were filled with various liquids. IL-6 and IL-1β mRNA expression was increased in rabbit esophageal mucosa stimulated with acid. Chinese herbal medicine significantly reduced the levels of IL-6 and IL-1β mRNA expression in the in vitro cultured rabbit esophageal mucosa. Using Chinese herbal medicine to treat in vitro models of RE, we found that the IL-6 and IL-1β mRNA expression levels went down, near to or lower than the normal control levels, compared with the group treated with acidified buffer solution.

CONCLUSIONSChai Lai Prescription lowered the IL-1β and IL-6 cytokine mRNA levels and protected the esophageal mucosa in the in vitro models of reflux esophagitis, suggesting that the traditional Chinese herbal compound may be able to treat reflux esophagitis by inhibiting the its inflammatory mediators.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Esophagitis, Peptic ; drug therapy ; metabolism ; Interleukin-1beta ; genetics ; Interleukin-6 ; genetics ; Male ; Rabbits

OBJECTIVETo observe the intervention effect of acupuncture on early onset of selec- tive serotonin reuptake inhibitors (SSRIs) in treating depressive disorder, and to study its effect on ser- um 5-HT and unbalanced inflammatory cytokines secreted by TH1/TH2.

METHODSTotally 90 patients with depressive disorder were randomly assigned to the drug control group (as the control group, 45 cases) and the acupuncture combined drug treatment group (as the treatment group, 45 cases). All patients were treated for 4 consecutive weeks. Another 45 healthy subjects were recruited as a healthy control group. The effect of acupuncture on early onset of SSRls in treating acute phase depressive disorder pa- tients was evaluated by HAMD score in the control group and the treatment group before treatment,and at weekends of the 1st, 2nd, and 4th week after treatment. Besides, their serum levels of 5-HT, IL-1β and IL-6 (secreted by TH1), and IL-4 and IL-10 (secreted by TH2) were detected before treatment and after treatment at the weekend of the 4th week.

RESULTSCompared with the healthy control group,serum lev- els of 5-HT, IL-4, and IL-10 decreased in the two drug-treated groups before treatment (P < 0.01); serum levels of IL-1β and IL-6 increased (P <0.01). Compared with before treatment in the same group, HAMD score decreased in the control group at weekends of the 2nd and the 4th week after treatment (P < 0.01); HAMD scores decreased in the treatment group at weekends of the 1st, 2nd, 3rd,and 4th week after treatment (P < 0.01); serum levels of 5-HT, IL-4, and IL-10 increased,serum levels of IL-1β and IL- 6 decreased in the two drug-treated groups after treatment (all P < 0.01). Compared with the control group at the same time point,HAMD scores decreased in the treatment group at weekends of the 1st, 2nd,3rd,and 4th week after treatment (P < 0.01),serum levels of 5-HT, IL-4, and IL-10 increased (P < 0.05, P < 0.01), serum levels of IL-6 decreased (P < 0. 01).

CONCLUSIONAcupuncture could accelerate early onset of SSRIs in treating acute phase depressive disorder, and effectively regulate serum 5-HT levels and inflammatory cytokines secreted by TH1/TH2.

Acupuncture Therapy ; Cytokines ; Depressive Disorder ; drug therapy ; Drugs, Chinese Herbal ; Humans ; Interleukin-10 ; Interleukin-1beta ; Interleukin-4 ; Interleukin-6 ; Serotonin Uptake Inhibitors ; therapeutic use

OBJECTIVETo explore the effect of combination therapy of tetramethylpyrazine (TMP) with methotrexate (MTX) on collagen induced arthritis (CIA) rats.

METHODSTotally 55 male SD rats were stratified by body weight. Nine of them were randomly recruited as the normal control group. The rest 46 were immunized with type II bovine collagen (C II) for establishing rheumatoid arthritis (RA) model. Forty successfully modeled rats were randomly divided into 4 groups according to swollen toe degree, i.e., the CIA group, the TMP group, the MTX group, and the TMP plus MTX group, 10 in each group. Rats in the MTX group were administered with MTX (1. 2 mg/kg) , once per week for 4 continuous weeks. Those in the TMP group were administered with 40 mg/kg TMP, once per day for 10 continuous days, and then discontinued for 7 successive days, and continued for another 10 successive days. Rats in the TMP plus MTX group were administered with a mixture of equal dose MTX and TMP, and when MTX was discontinue, TMP was administered according to the way in the TMP group. Equal volume of saline solution was given to rats in the normal control group and the CIA group. Clinical parameters including ankle width (mediolateral diameter) and hindpaw swelling were measured at day 0, 4, 11, 18, and 26 after treatment. Rats were sacrificed 28 days after treatment, their knee joints and ankle joints were collected for pathological analyses. Serum levels of IL-1β, IL-6, and IL-17A were detected by ELISA. Changes of fibrinogen (FIB) and platelet aggregation rate (PAg) were detected.

RESULTSCompared with the normal control group, the ankle width and hindpaw swelling increased significantly (P < 0.01), contents of FIB and PAg increased obviously (P < 0.05, P < 0.01), serum levels of IL-1β, IL-6, and IL-17 increased remarkably (P <0. 01) in the CIA group. Obvious cell proliferation, inflammatory cell infiltration, hyperemia and edema of synovial tissues could be seen. Pannus formed and immerged in cartilages, resulting in necrosis. Compared with the model group, changes of ankle width and hindpaw swelling were all alleviated in each medicated group (P <0. 05, P <0. 01). Of them, the effect was superior in the MTX group to that of the TMP group and the MTX plus TMP group (P < 0.05, P < 0.01). Contents of FIB, serum levels of IL-1β and IL-6 decreased significantly in the MTX group (P < 0.05). Contents of FIB, serum levels of IL-1β and IL-6 decreased significantly in the TMP group and the MTX plus TMP group (P < 0.05). Besides, serum levels of FIB and IL-6 were obviously lower in the MTX plus TMP group than in the TMP group and the MTX group (P < 0.01). Levels of PAg and IL-17A were more significantly lowered in the TMP group than in the MTX plus TMP group and the MTX group. Pathological changes could be alleviated in each medicated group, with the optimal effect obtained in the MTX plus TMP group.

CONCLUSIONCombination of TMP with MTX could significantly ameliorate inflammatory reactions and FIB contents of CIA rats.

Animals ; Arthritis, Experimental ; Arthritis, Rheumatoid ; Cattle ; Collagen Type II ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Hemorheology ; Interleukin-17 ; Interleukin-1beta ; Interleukin-6 ; Male ; Methotrexate ; therapeutic use ; Pyrazines ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Synovial Membrane

OBJECTIVETo investigate the effects of prostaglandin E1 (PGE1) on the cytokines (IL-6, IL-8, IL-10) levels and ischemic -reperfusion injury of myocardium during cardiac surgery.

METHODSA total of 30 patients undergoing cardiac surgery (mitral valve replacement) under extracorporeal circulation were randomized into two groups: PGE1 group (receiving 0.04 microg.kg(-1). min(-1) of Lipo-PGE1 from the beginning of surgery to the end of study, n=15) and control group (no PGE1 given, n=15). Levels of serum IL-6, IL-8 and IL-10 were measured by enzyme-linked immunosorbent assays (ELISA). Isoenzyme of creatine kinase with muscle and brain subunits(CKMB) and troponin-T (cTn-T) were measured by ultraviolet absorption spectrophotometry method and enzyme immunoassay on 5 time-points during the study.

RESULTIn both groups serum IL-6, IL-8, CK-MB and cTn-T levels increased significantly after aorta declamping (especially from 2 h after aorta declamping) compared with preoperative levels (P<0.05).However,the elevations of these cytokines and enzymes were more prominent in the control group than in the PGE1 group (P<0.05). Serum IL-10 concentration increased significantly from 2 h after aorta declamping compared with preoperative value (P<0.05); but there were no differences between the two groups. IL-6 and IL-8 levels were correlated with CK-MB and cTn-T concentrations (r2=0.40, r2=0.38 P>0.05 and r2=0.56, r2=0.14; P>0.05, respectively).

CONCLUSIONDuring cardiac surgery (mitral valve replacement) PGE1 may suppressed the production of IL-6, IL-8 but not IL-10, which may be related to its myocardial protection effect.

Alprostadil ; therapeutic use ; Humans ; Interleukin-10 ; blood ; Interleukin-6 ; blood ; Interleukin-8 ; blood ; Mitral Valve ; surgery ; Myocardial Reperfusion Injury ; prevention & control ; Rheumatic Heart Disease ; surgery ; Troponin T ; blood

OBJECTIVETo investigate the protective effect of mannitol therapy on the vital organs and explore the underlying mechanisms in New Zealand rabbits with severe burn injury.

METHODSTwelve New Zealand rabbits with severe burn injury (30% of TBSA) were randomized to receive fluid resuscitation with saline (control) or mannitol therapy starting at 1 h after the injury. Serum and urine samples were collected before and at 1, 4, 8, 24, and 48 h after the injury for detection of TNF-α, IL-6, ALT, AST, GGT, CK, CK-MB, BUN and Cr levels using sandwich ELISA.

RESULTSOne hour after sever burn injury, the serum levels of TNF-α and IL-6 began to increase along with ALT, AST, GGT, CK, CK-MB, BUN and Cr levels. Compared with control group, the rabbits in mannitol group showed significantly higher 48 h urine excretion of TNF-α (145 ± 8 vs 78 ± 1 0 pg/ml, P<0.05) and IL-6 (93 ± 6 vs 40 ± 8 pg/ml, P<0.05) but with lowered serum levels of TNF-α (0.62 ± 0.02 vs 0.83 ± 0.02 pg/ml, P<0.05) and IL-6 (0.45 ± 0.03 vs 0.56 ± 0.03 pg/ml, P<0.05) as well as lowered serum ALT, AST, GGT, CK, CK-MB, BUN and Cr levels (P<0.05).

CONCLUSIONIn rabbits with severe burn injury, mannitol therapy can decrease serum TNF-α and IL-6 levels early after the injury to ameliorate potential functional impairment of the heart, liver and kidneys.

Animals ; Burns ; blood ; drug therapy ; Fluid Therapy ; Interleukin-6 ; blood ; Male ; Mannitol ; therapeutic use ; Rabbits ; Tumor Necrosis Factor-alpha ; blood

Adult ; Antidepressive Agents ; therapeutic use ; Bipolar Disorder ; blood ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Interferon-gamma ; blood ; Interleukin-1 ; blood ; Interleukin-6 ; blood ; Male ; Middle Aged ; Perylene ; analogs & derivatives ; therapeutic use ; Phytotherapy ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the therapeutic effect of Qingyi Decoction (QYD) and tetrandrine (Tet), used singly or combind, in treating miniature pigs with severe acute pancreatitis (SAP) and its mechanism.

METHODSThirty-two Guizhou miniature pigs were made into SAP model by pancreatic duct retrograde injection of 5% sodium taurocholate. They were randomly divided into 4 groups: the control group, the QYD group, the Tet group and the combined treated group. The serum amylase activity and interleukin-1 and 6 (IL-1, IL-6) contents in serum from vena cava and portal vein were tested by biochemistry and radioimmunoassay (RIA). Serum emodin and plasma Tet levels were measured by high performance liquid chromatography (HPLC) 24, 48 and 72 hrs after treatment. And the pathological changes of pancreas, lung and liver were observed under microscope.

RESULTSThe mortality of SAP pigs was reduced significantly and the inflammatory injury of the organs was ameliorated obviously in all treated groups, and the increased amylase activity and IL-1, IL-6 levels was attenuated. The therapeutic effect was much more obvious, and the plasma Tet level at different time points were much higher in the combined treated group than those in the other two groups treated by single drug (P < 0.01).

CONCLUSIONBoth QYD and Tet could treat effectively SAP through multiple pathways, combination of them reveals an elevation of serum drug concentration and shows a synergistic effect.

Alkaloids ; blood ; pharmacokinetics ; therapeutic use ; Animals ; Benzylisoquinolines ; blood ; pharmacokinetics ; therapeutic use ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacokinetics ; therapeutic use ; Emodin ; blood ; Female ; Interleukin-1 ; blood ; Interleukin-6 ; blood ; Male ; Pancreatitis, Acute Necrotizing ; blood ; drug therapy ; Phytotherapy ; Random Allocation ; Swine, Miniature

Dendrobium officinale can serve as Chinese medicinal material effective in nourishing yin, clearing heat, and producing fluid, and is used to treat throat diseases, but its active substances and mechanism are not clear. To clarify the active fraction and underlying mechanism of D. officinale against chronic pharyngitis(CP), the present study induced a CP model in rats by pepper water combined with low-concentration ammonia, and crude polysaccharides of D. officinale(DOP), non-polysaccharides of D. officinale(DON), and total extract of D. officinale(DOT)(0.33 g·kg~(-1), calculated according to the crude drug) were administered by gavage for six weeks. The changes in oral secretions and pharyngeal conditions of rats with CP were observed and rated. The hematological indicators were determined by an automatic hematology analyzer. The serum levels of pro-inflammatory factors, such as tumor necrosis factor-alpha(TNF-α), interleukin 1β(IL-1β), and interleukin 6(IL-6), and T-lymphocyte cytokines, including interferon γ(IFN-γ), interleukin 4(IL-4), interleukin 17(IL-17), and transforming growth factor β1(TGF-β1) were detected by the enzyme-linked immunosorbent assay(ELISA). The proportions of CD3~+, CD4~+, and CD8~+cells in peripheral blood T lymphocyte subsets were determined by the flow cytometry. The histomorphological changes of the pharynx were observed by hematoxylin-eosin(HE) staining. The protein expression of nuclear factor-κB P65(NF-κB P65), cyclooxygenase-2(COX-2), F4/80, and monocyte chemoattractant protein-1(MCP-1) in the pharynx were detected by immunohistochemistry and Western blot. The results showed that DOP and DON could significantly relieve pharyngeal lesions, reduce white blood cells(WBC) and lymphocytes(LYMP), decrease the levels of pro-inflammatory factors TNF-α, IL-6, and IL-1β, and inhibit the protein expression of NF-κB P65, COX-2, F4/80, and MCP-1 in the pharynx. DOP was superior in reducing oral secretions and serum IL-17 level and inferior in increasing CD4~+/CD8~+ratio to DON. It is suggested that both polysaccharides and non-polysaccharides of D. officinale have anti-PC effects and the anti-inflammatory mechanism may be related to the regulation of T lymphocyte distribution and inhibition of the inflammatory signaling pathways mediated by NF-κB P65. The anti-inflammatory effect of DOP may be related to the regulation of Th17/Treg balance, while that of DON may be related to the regulation of the Th/Tc ratio.
Ammonia/therapeutic use* ; Animals ; Anti-Inflammatory Agents/therapeutic use* ; Cyclooxygenase 2 ; Dendrobium/chemistry* ; Interleukin-17/therapeutic use* ; Interleukin-6 ; NF-kappa B/metabolism* ; Pharyngitis/drug therapy* ; Plant Extracts/chemistry* ; Polysaccharides/pharmacology* ; Rats ; Tumor Necrosis Factor-alpha ; Water