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.     

盐酸坦索罗辛联合吲哚美辛栓治疗输尿管下段结石的初步研究

目的评价盐酸坦索罗辛与吲哚美辛栓联合治疗输尿管下段结石的临床疗效。方法 89例确诊为单纯输尿管下段结石(结石均小于8mm)的患者,随机分为三组:试验组,口服盐酸坦索罗辛(0.4mg/d)和吲哚美辛栓0.1g塞肛,1次/d;对照组1,口服中药排石颗粒4g/次,2次/d;对照组2,口服盐酸坦索罗辛0.4mg,1次/d;观察结石排出率、排出时间、肾绞痛发生率、药物不良反应发生率、干预治疗率等。结果试验组结石排出率为90.63%(29/32),对照组1和对照组2结石排除率分别为42.31%(11/26)和70.97%(22/31),试验组与对照组1比较,试验组与对照组2比较,两个对照组比较,差异均有统计学意义(χ2分别=4.76、3.94、15.65;P均<0.05)。试验组的结石排出时间为(8.73±2.15)d,对照组1和对照组2结石排出时间分别为(12.31±1.52)d和(9.52±1.61)d,试验组与对照组1比较,试验组与对照组2比较,两个对照组比较,差异均有统计学意义(t分别=6.77、-1.70、-7.35;P均<0.05)。试验组、对照组1、照组2的肾绞痛发生率分别为31.25%(10/32)、63.28%(17/26)、45.16%(14/31),试验组和对照组2以及对照组1和对照组2比较,差异均无统计学意义(χ2分别为1.29、2.33,P均>0.05);试验组和对照组1比较,差异有统计学意义(χ2=8.24,P<0.05)。三组患者均未发生严重的药物不良反应(χ2分别=0.33、0.23、0.35,P均>0.05)。结论联合应用盐酸坦索罗辛和吲哚美辛栓治疗输尿管下段结石安全、副作用小,能明显提高排石率,缩短排石时间,并能减少镇痛药物的应用。     

Complementarity of in vitro and in vivo models for the evaluation of gastro‐protective effects of pharmacological substances

Gastric mucosa is frequently exposed to various gastric irritants, and there is a continuing requirement to develop new gastro‐protective agents. This study compares the effects of three such agents, sucralfate, rebamipide, and cimetidine in both in vivo and in vitro indomethacin‐induced gastric damage models. For the in vivo approach, rats were orally administered sucralfate, rebamipide, and cimetidine at 300 mg/kg before an acute dose of indomethacin (30 mg/kg). Gastric lesions were then macroscopically examined. For the in vitro approach, gastric mucosal cells were incubated with sucralfate (3 and 5 mg/mL), rebamipide (0.3 and 1 mm ), and cimetidine (10 and 50 μg/mL) before exposure to indomethacin (3.8 mm ). The release of lactate dehydrogenase (LDH) and mitochondrial function were then measured. Sucralfate, rebamipide, and cimetidine displayed gastro‐protective effects in vivo (decreased number of gastric ulcers: ?50% P < 0.05, ?22% NS, and ?69% P < 0.05, respectively, and reduced length of gastric lesions: ?62% P < 0.05, ?29% NS, and ?70% P < 0.001, respectively). Cell damage induced by indomethacin in vitro was inhibited by sucralfate (LDH release) and by rebamipide and cimetidine (mitochondrial function and LDH release). In contrast, sucralfate accentuated the indomethacin‐induced decrease in mitochondrial function. Although cultured gastric cells offer a promising tool for evaluating the cytotoxic or protective effects of test compounds, data from in vivo models are still needed to confirm in vitro data. Using both approaches provides more comprehensive insight into the effects of test compounds on the gastric mucosa.     

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).