Section 1 The effect of ibuprofen on lymphocyte stimulation by phytohaemagglutillin in Vitro

Summary

A study was carried out to determine the effects of ibuprofen, phenylbutazone and indomethacin at different concentrations on phytohaemagglutinin-induced stimulation of lymphocytes in vitro. The results indicate that all three drugs inhibit lymphocyte stimulation, and at concentrations achieved by ibuprofen and phenylbutazone in vivo. It may be, therefore, that part of the effect of ibuprofen in rheumatoid arthritis is due to inhibition of lymphocyte function.     

Indomethacin and Aspirin: Effect of Nonsteroidal Anti-Inflammatory Agents on the Rate of Fracture Repair in the Rat

A total of 210 male Charles River CD rats, 45 days old, were subjected to a fracturing of the right radius and ulna by digital pressure while under ether anesthesia. These animals were then assigned randomly to dose groups (1, 2 or 4 mgAg/day of indomethacin and 100, 200, or 300 mgAg/day of aspirin) and were dosed for 21 days. Dose levels were chosen to provide approximately equipotent levels of the test compounds with the highest dose approaching toxicity. Radiographs were taken of control-rat fractures on days 8, 14, and 21. Three rats at 4 mgAg/day of indomethacin died of intestinal perforation prior to scheduled sacrifice. on day 22, all remaining rats were sacrificed by exsanguination under anesthesia. Histologic sections of the radius and ulna were examined in random sequence without knowledge of the treatment regimen. A histologic grade based on the morphologic stage of fracture healing was given. There was a drug- and dose-related retardation of fracture healing, which was statistically significant at all dose levels of indomethacin and the highest level of aspirin, as compared to controls. Decreased mean grades in the groups given 100 and 200 mgAg/day of aspirin, though not statistically significant, suggest a retarding effect on fracture healing at these levels also. No statistically significant changes in numbers of pseudoarthroses were found.     

Pharmacological approach to acute pancreatitis

The aim of the present review is to summarize the current knowledge regarding pharmacological prevention and treatment of acute pancreatitis （AP） based on experimental animal models and clinical trials. Somatostatin （SS） and octreotide inhibit the exocrine production of pancreatic enzymes and may be useful as prophylaxis against Post Endoscopic retrograde cholangiopancreatography Pancreatitis （PEP）. The protease inhibitor Gabexate mesilate （GM） is used routinely as treatment to AP in some countries, but randomized clinical trials and a meta-analysis do not support this practice. Nitroglycerin （NGL） is a nitrogen oxide （NO） donor, which relaxes the sphincter of Oddi. Studies show conflicting results when applied prior to ERCP and a large multicenter randomized study is warranted. Steroids administered as prophylaxis against PEP has been validated without effect in several randomized trials. The non-steroidal anti-inflammatory drugs （NSAID） indomethacin and diclofenac have in randomized studies showed potential as prophylaxis against PEP. Interleukin 10 （IL-10） is a cytokine with anti-inflammatory properties but two trials testing IL-10 as prophylaxis to PEP have returned conflicting results. Antibodies against tumor necrosis factor-alpha （TNF-α） have a potential as rescue therapy but no clinical trials are currently being conducted. The antibiotics beta- lactams and quinolones reduce mortality when necrosis is present in pancreas and may also reduce incidence of infected necrosis. Evidence based pharmacological treatment of AP is limited and studies on the effect of potent anti-inflammatory drugs are warranted.     

Effect of indomethacin on Bfl‐1, WISP‐1 and proliferating cell nuclear antigen in colon cancer cell line HCT116 cells

BACKGROUND: Non‐steroidal anti‐inflammatory drugs such as indomethacin can inhibit the growth of tumors through both the cyclooxygenase‐2 (COX‐2) dependant and COX‐2 independent pathways, but the exact mechanism has not yet been shown. In our previous study, COX‐2 independent proteins (Bfl‐1, WISP‐1 and proliferating cell nuclear antigens [PCNA]) in indomethacin‐treated colorectal cancer cells with the use of proteomics technology had been identified. OBJECTIVES: To study and confirm the effect of indomethacin on the expression of Bfl‐1, WISP‐1 and PCNA in human colon cancer line HCT116 cells and the COX‐2 independent tumor inhibiting pathway. METHODS: Human colon cancer cell line HCT116 cells were divided into a treatment with indomethacin (IC 50) group, and a treatment with dimethyl sulfoxide (DMSO) as a control group for 48 h. The expression of Bfl‐1, WISP‐1 and PCNA, mRNA and protein were determined by a real‐time quantitative PCR and Western blot, respectively. RESULTS: Indomethacin down‐regulated the expression of Bfl‐1, WISP‐1 and PCNA mRNA in vitro (9.53 ± 0.15 vs 27.87 ± 0.12, 7.37 ± 0.58 vs 20.17 ± 0.58, 5.17 ± 0.06 vs 0.87 ±  0.06). Indomethacin also down‐regulated the expression of Bfl‐1, WISP‐1 and PCNA protein (40.01 ± 1.61 vs 43.76 ± 1.63, 22.50 ± 1.17 vs 30.30 ± 1.55, 17.69 ± 1.18 vs 20.80 ± 1.08). CONCLUSIONS: Inducing apoptosis and inhibiting proliferation contribute to the anticancer activity of indomethacin via COX‐2 independent pathway of Bfl‐1, WISP‐1 and PCNA. This further confirms the results of our previous study.     

Effect of prolactin and prostaglandins on the stimulation of prolactin binding sites in the male rat liver

Induction of prolactin hepatic receptors in the male rat by exogenous prolactin and the possible involvement of prostaglandins in this process were studied. When ovine prolactin 500 microgram/kg was administered sc in saline containing 10% PVP (polyvinylpyrrolidone), twice daily for 6.5 days and the rats killed 48 h after the last injection, the specific binding of 125I-labelled ovine prolactin to prolactin hepatic receptors was raised 26-fold, while administration of prolactin in saline only caused a 7-fold increase. A well correlated log dose-response relationship was demonstrated between 12.5 and 500 microgram of prolactin in saline-PVP, with lowest dose causing an 18-fold increase in binding. A shorter 4.5 day treatment of prolactin in saline-PVP caused only a small 3-fold increase in prolactin binding. Scatchard analysis showed that these increases resulted from increases in receptor concentration. The effect of prolactin on the induction of the hepatic receptors could not be mimicked by PGE1, PGE2 or PGF2 alpha, nor could PGF2 alpha synergize with a short treatment with prolactin. Further, indomethacin caused no significant effect on this action of prolactin. It seems that prolactin does not induce its own receptors in the rat liver by stimulation of prostaglandins in this tissue.     

Intravenous nucleosides and a nucleotide promote healing of small bowel ulcers in experimental enterocolitis

Our aim was to evaluate the possible beneficial effect of intravenous nucleosides and a nucleotide in healing small bowel ulceration in a rat model of enterocolitis. Fourteen Lewis female rats were randomized into total parenteral nutrition (TPN,N=7) and TPN + nucleosides and a nucleotide (NS/NT,N=7) groups. After adaptation, two doses of indomethacin (7.5 mg/kg) were administered subcutaneously 24 hr apart to each animal in both groups. Concomitant with the first dose of indomethacin, TPN or TPN + NS/NT·were infused for four days. The TPN and TPN + NS/NT were isocaloric and isonitrogenous. At the end of four days, total ulcer length in the entire small bowel was measured. The mucosa surrounding ulcers was studied by optical microscopy. Immunohistochemistry was performed for proliferating cell nuclear antigen (PCNA). Ileal crypt and villus lengths were measured with an eyepiece micrometer, crypt-villus ratios were calculated, and crypt mitotic index and percentage of PCNA-labeled cells determined to assess cellular proliferation. Total ulcer length decreased significantly in the TPN + NS/NT group compared to the TPN group (42 vs 76 mm). In the TPN + NS/NT versus TPN group, the ileal mucosa surrounding ulcers showed significantly greater crypt length (21%) and there was increased crypt-villus ratio (0.53 vs 0.39), crypt mitotic index (1.2 vs 0.9), and PCNA labeling (43% vs 30%). We conclude that in rats with indomethacin-induced enterocolitis, administration of TPN + NS/NT for four days resulted in significant healing of small bowel ulcers, as indicated by decreased ulcer length. This effect of NS/NT appears to relate, in part, to increased cell proliferation, evidenced by increased crypt length, crypt-villus ratio, mitotic index, and PCNA labeling.     

Suppressive effects of nonsteroidal anti-inflammatory drugs diclofenac sodium,salicylate and indomethacin on delayed rectifier K+-channel currents in murine thymocytes

Lymphocytes predominantly express delayed rectifier K⁺-channels (Kv1.3) in their plasma membranes, and the channels play crucial roles in the lymphocyte activation and proliferation. Since nonsteroidal anti-inflammatory drugs (NSAIDs), the most commonly used analgesic and antipyretic drugs, exert immunomodulatory effects, they would affect the channel currents in lymphocytes. In the present study, employing the standard patch-clamp whole-cell recording technique, we examined the effects of diclofenac sodium, salicylate and indomethacin on the channel currents in murine thymocytes and the membrane capacitance. Diclofenac sodium and salicylate significantly suppressed the pulse-end currents of the channel. However, indomethacin suppressed both the peak and the pulse-end currents with a significant increase in the membrane capacitance. This study demonstrated for the first time that NSAIDs, such as diclofenac sodium, salicylate and indomethacin, exert inhibitory effects on thymocyte Kv1.3-channel currents. The slow inactivation pattern induced by indomethacin was thought to be associated with microscopic changes in the plasma membrane surface detected by the increase in the membrane capacitance.     

Interleukin-1 receptor antagonist does not prevent endotoxin-induced inhibition of gastric acid secretion in rats

The underlying mechanisms involved in endotoxin-induced inhibition of gastric acid secretion were investigated in conscious rats with pylorus ligation for 2 hr. Intraperitoneal injection of endotoxin (0.1, 1, and 5 µg/rat) inhibited gastric acid output by 31%, 80%, and 84% respectively. Intraperitoneal endotoxin (1 µg/rat) -induced inhibition of gastric acid secretion was not altered by pretreatment with the interleukin-1 receptor antagonist, IL-1RA, indomethacin, naloxone, or capsaicin. Treatments were injected peripherally at doses previously shown to antagonize the antisecretory effect of exogenous interleukin-1, to inhibit prostaglandin synthesis in the stomach and brain, to block opiate receptors, and to alter functioning of unmyelinated afferent nerve fibers. These results indicate that the antisecretory effect of endotoxin can be expressed by factors other than interleukin-1, prostaglandins, or opioid peptides that do not require the integrity of capsaicin-sensitive afferent pathways.Supported by the National Institute of Arthritis Metabolism and Digestive Disease, grant DK-30110, and the Institute of Mental Health, grant MH-00663, a Research Grant from the Psychoneuroimmunology Program at UCLA, and the Direction General de Investigacion Cientificas Y Técnicas (DGICYT PM—0124 to Dr. E. Saperas).     

环氧合酶-2抑制剂对大鼠骨关节炎模型软骨中血管内皮生长因子表达的影响

目的研究环氧合酶-2抑制剂(吲哚美辛)对大鼠骨关节炎模型关节软骨中血管内皮生长因子(VEGF)表达的影响。方法健康雄性wistar大鼠30只,随机均分为对照组、骨性关节炎组(OA组)和吲哚美辛处理组。利用膝关节注射4%的木瓜蛋白酶溶液的方法制作骨关节炎模型。采用关节炎评分法评定各组大鼠平均关节炎指数(MAI),免疫组化方法与Western blot方法检测关节软骨中VEGF蛋白的表达变化。结果 OA模型组随着造模的时间延长,关节出现了重度红肿现象,对照组关节无任何异常变化;与OA模型组比较,吲哚美辛组的关节炎症反应呈现消退现象。VEGF蛋白在对照组大鼠关节软骨仅有微量表达,而在术后8周OA组大鼠关节软骨表达则显著升高(P0.01);与OA组相比,吲哚美辛组大鼠关节软骨中VEGF蛋白表达显著降低(P0.01)。结论吲哚美辛对骨关节炎的抑制作用可能与下调VEGF蛋白的表达相关。     

自体血注射联合吲哚美辛巴布膏治疗顽固性网球肘的临床疗效观察

目的观察自体血注射联合吲哚美辛巴布膏治疗顽固性网球肘的临床疗效。方法顽固性网球肘患者90例,采用数字表法将90例患者随机分为观察组(45例,自体血注射联合吲哚美辛巴布膏)和对照组(45例,局部封闭)。观察两组患者治疗前及治疗后不同时期疼痛程度和肘关节功能变化。采用视觉模拟评分法(visual analogue scale,VAS)、Mayo肘关节功能评分(Mayo elbow performance score,MEPS)和Nirschl分级评分评定治疗效果。结果观察组治疗后VAS评分、Nirschl分级评分和MEPS评分T1、T2和T3与治疗前T0比较,差异有统计学意义(P0.05)。对照组治疗后VAS评分T1、T2与治疗前T0比较,差异有统计学意义(P0.05);Nirschl分级评分T1和T2与治疗前T0比较,差异有统计学意义(P0.05);MEPS评分T1与治疗前T0比较,差异有统计学意义(P0.05)。观察组治疗后VAS评分、Nirschl分级评分和MEPS评分T2、T3与对照组T2、T3比较,差异有统计学意义(P0.05)。结论两组患者近期疗效均稳定,自体血注射联合吲哚美辛巴布膏治疗顽固性网球肘的中远期疗效明显优于局部封闭治疗。