Flavocoxid is as effective as naproxen for managing the signs and symptoms of osteoarthritis of the knee in humans: a short-term randomized,double-blind pilot study

Flavocoxid (Limbrel), a proprietary mixture of flavonoid molecules (baicalin and catechin), was tested against a traditional nonsteroidal anti-inflammatory drug, naproxen, for the management of the signs and symptoms of moderate osteoarthritis (OA) in humans. Discomfort and global disease activity were used as the primary end points, and safety assessments were also taken for both treatments as a secondary endpoint. In this double-blind study, 103 subjects were randomly assigned to receive either flavocoxid [500 mg twice daily (BID)] or naproxen (500 mg BID) in a 1-month onset of action trial. Outcome measures included the short Western Ontario and McMaster University Osteoarthritis Index, subject Visual Analogue Scale for discomfort and global response, and investigator Visual Analogue Scale for global response and fecal occult blood. Both flavocoxid and naproxen showed significant reduction in the signs and symptoms of knee OA (P ≤ .001). There were no statistically detectable differences between the flavocoxid and naproxen groups with respect to any of the outcome variables. Similarly, there were no statistically detectable differences between the groups with respect to any adverse event, although there was a trend toward a higher incidence of edema and nonspecific musculoskeletal discomfort in the naproxen group. In this short-term pilot study, flavocoxid was as effective as naproxen in controlling the signs and symptoms of OA of the knee and would present a safe and effective option for those individuals on traditional nonsteroidal anti-inflammatory drugs or cyclooxygenase-2 inhibitors. A low incidence of adverse events was reported for both groups.     

Immunohistochemical localization of prostaglandin H synthase isoenzyme proteins in the gingival tissue of patients with periodontitis

Prostaglandins are inflammatory mediators that are believed to play an important role in the pathophysiology of periodontal disease. Prostaglandin H synthase (PGHS, EC 1.14.99.1) is the rate-limiting enzyme in prostaglandin biosynthesis. The enzyme exists as two separately encoded isoforms. PGHS-1 which is constitutively expressed and PGHS-2 which is induced by inflammatory stimuli. This is the first report describing the expression of the isoenzymes in gingival tissue from patients diagnosed with adult periodontitis. Tissue was fixed in an alcohol-based fixative and embedded in paraffin. Methods were developed using immunohistochemical controls, such that embedded sections could be processed and stained using isoform-specific antibodies and a peroxidase-antiperoxidase immunohistochemistry technique. Along with populations of mononuclear inflammatory cells, endothelial cells and fibroblasts, the gingival epithelial cell layer appears to be a rich and important source of prostaglandin production within the periodontium of patients with periodontitis as detected by this newly developed immunohistochemical staining technique for PGHS-1 and PGHS-2.     

Placentally derived prostaglandin E2 acts via the EP4 receptor to inhibit IL-2-dependent proliferation of CTLL-2 T cells

A number of immunomodulatory molecules are present in the placenta, including cytokines, prostaglandins, progesterone and indoleamine 2,3-dioxygenase. An undefined factor capable of down-regulating T-cell activity has recently been reported [1] as being produced by short-term cultures of placental fragments. By careful repetition of these studies we have confirmed that chorionic villi isolated from term placenta produce a low molecular weight, heat stable factor capable of inhibiting the IL-2-dependent proliferation of mouse CTLL-2 cells. This activity was not due, however, to a previously unknown immunosuppressive molecule, but rather to prostaglandin E2 (PGE2). Expression of cyclooxygenase (COX)-2 was detected in the syncytiotrophoblast of chorionic villi explants using immunohistochemistry. Culture of the explants in the presence of the COX-1/COX--2 inhibitors indomethacin and diclofenac, or with the COX-2-selective inhibitor DFP, blocked the production of the immunosuppressive factor. The immunosuppressive activity was restored by adding PGE2 to the supernatants obtained from diclofenac-inhibited explants. A number of different receptors are involved in mediating the biological effects of prostaglandins. By utilizing selective antagonists of individual receptors, we have established that the immunosuppressive effect of PGE2 on CTLL-2 cells is exerted via the EP4 receptor. Thus, addition of an EP4-selective antagonist, but not of EP1 or EP3 antagonists, abolished the immunosuppressive effect of PGE2 on CTLL-2 cells. This may have implications for attempts to selectively manipulate T-cell responses.     

Immunohistochemical characteristics of duodenal adenomas in familial adenomatous polyposis with special reference to cell kinetics

The duodenum is the second most frequent site of cancer in patients with familial adenomatous polyposis (FAP). The main objective of this study was to evaluate the cell kinetics in duodenal and ampullary adenomas in FAP. The endoscopic and biopsy findings of duodenal adenomas in 22 FAP subjects and 18 non-FAP subjects were compared. Adenomas in FAP included 15 ampullary adenomas and 17 nonampullary adenomas. The cell kinetics was evaluated by immunohistochemistry for Ki-67, p53, bcl-2, and cyclooxygenase 2 (COX2), and the apoptotic index (AI) as determined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) method. Any correlations between the indices for cell kinetics and the endoscopic findings were identified. All 50 adenomas were histologically verified to be tubular adenoma with low-grade dysplasia. Neither the expression of Ki-67, p53, bcl-2, and COX2 nor the AI differed substantially between FAP and non-FAP subjects. In patients with FAP, duodenal adenoma tended to have a higher Ki-67-labeling index than the ampullary adenoma (54.3 +/- 11.3 versus 46.8 +/- 12.7; .05 < P < .1). In addition, the Ki-67-labeling index in endoscopically normal or slightly enlarged ampullary adenoma was significantly higher than that in markedly enlarged ampullary adenoma (51.8 +/- 11.4 versus 39.4 +/- 11.3; P < .05). Duodenal adenoma in FAP subjects was not found to have a higher proliferative activity or a smaller degree of apoptosis compared with those in non-FAP subjects. The smaller proliferative activity in larger ampullary adenoma may thus be related to the static nature of ampullary adenoma in FAP.     

人颈动脉粥样硬化斑块环氧化物酶-2及Ⅰ型前列腺素合成酶的表达

目的探讨环氧化物酶-2(cyclooxygenase type 2,COX-2)及Ⅰ型前列腺素合成酶(membrane associated prostaglandin E-1,mPGES-1)在人颈动脉粥样硬化斑块中的表达变化及作用机制。方法收集24例人颈动脉粥样硬化斑块标本和10例肠系膜动脉标本做对照组,应用免疫组织化学及逆转录PCR方法测定COX-2及mPGES-1mRNA表达水平,Western印记方法检测COX-2及mPGES-1的蛋白表达水平。比较不同程度动脉粥样硬化组织间COX-2、mPGES-1 mRNA表达水平及蛋白表达水平。结果颈动脉粥样硬化斑块组的免疫组织化学染色检测COX-2和mPGES-1呈阳性表达,斑块组COX-2 mRNA和mPGES-1 mRNA表达与对照组相比上调,差异有统计学意义(P<0.05);COX-2及mPGES-1 mRNA上调水平相关(P<0.05);颈动脉粥样硬化斑块的COX-2蛋白表达上调水平与对照组相比差异有统计学意义(P<0.05);颈动脉粥样硬化斑块COX-2、mPGES-1 mRNA及蛋白表达水平与病理损害程度有关,差异有统计学意义(P<0.05)。结论COX-2及mPGES-1基因表达水平上调可能是进展性动脉粥样硬化损害的关键因素。     

Reduced expression of cyclooxygenase (COX) in idiopathic pulmonary fibrosis and sarcoidosis

AIMS: To test the hypothesis that cyclooxygenase (COX)-1 or COX-2 expression is defective in lungs in idiopathic pulmonary fibrosis (IPF) and to characterize the cellular distribution. IPF is a progressive inflammatory lung disorder with an adverse prognosis. Previous work has shown that prostaglandin E2 (PGE2) regulates collagen deposition and fibroblast proliferation and a defect in COX regulation may contribute to the fibrosis that occurs in IPF. METHODS: Immunohistochemistry was utilized to determine COX immunoreactivity in lung sections from 25 IPF, six sarcoidosis and 14 control subjects. RESULTS: COX-1 and COX-2 expression in bronchiolar epithelial cells was significantly lower in IPF and sarcoidosis than in controls. No significant difference was found in COX-2 expression between macrophages in IPF and control sections, but COX-2 was reduced in macrophages in sarcoidosis compared with controls. CONCLUSIONS: These studies confirm COX-2 loss in bronchial epithelial cells but not macrophages in IPF, and show for the first time reduced constitutive COX-1 expression in epithelial cells and macrophages. Similar abnormalities were observed in sarcoidosis.     

Brain prostaglandin formation is increased by α-synuclein gene-ablation during global ischemia

We have previously demonstrated that α-synuclein (Snca) gene ablation reduces brain arachidonic acid (20:4n − 6) turnover rate in phospholipids through modulation of endoplasmic reticulum-localized acyl-CoA synthetase activity. Although 20:4n − 6 is a precursor for prostaglandin (PG), Snca effect on PG levels is unknown. In the present study, we examined the effect of Snca ablation on brain PG level at basal conditions and following 30 s of global ischemia. Brain PG were extracted with methanol, purified on C₁₈ cartridges, and analyzed by LC–MS/MS. We demonstrate, for the first time, that Snca gene ablation did not affect brain PG mass under normal physiological conditions. However, total PG mass and masses of individual PG were elevated ∼2-fold upon global ischemia in the absence of Snca. These data are consistent with our previously observed reduction in 20:4n − 6 recycling through endoplasmic reticulum-localized acyl-CoA synthetase in the absence of Snca, which may result in the increased 20:4n − 6 availability for PG production in the absence of Snca during global ischemia and suggest a role for Snca in brain inflammatory response.     

CpG DNA induces cyclooxygenase-2 expression and prostaglandin production

Unmethylated CpG motifs found in bacterial DNA are potent activators of the innate and acquired immune systems, and rapidly induce the production of proinflammatory cytokines. We hypothesized that CpG DNA may also elicit the production of prostaglandins (PG), which are central lipid mediators of the immune and inflammatory response. To test our hypothesis, we stimulated murine spleen cells and RAW 264.7 murine macrophage cells with CpG DNA and assessed the effects on the PG synthesis pathway. Compared to control, DNA-containing CpG motifs induced >5-fold increase in PGE (2) production and rapidly up-regulated cyclooxygenase-2 (COX-2) at both the mRNA and protein level. CpG DNA was an extremely strong inducer of COX-2 as concentrations as low as 3 ng/ml induced COX-2 protein expression. The CpG DNA-induced PGE (2) down-regulated the immune response elicited by CpG. Blockade of PGE (2) production with selective COX-2 inhibitors or neutralizing anti-PGE (2) antibody markedly enhanced IFN-gamma secretion in vitro from CpG DNA-stimulated spleen cells. Moreover, selective COX-2 inhibition increased CpG DNA-induced IFN-gamma secretion in vivo. Inhibition of COX-2 also increased CpG DNA-induced lytic activity of NK cells. Taken together, these data indicate that DNA containing CpG motifs is a potent inducer of COX-2 and PGE (2) production. CpG-induced PG may subsequently down-regulate the immune and inflammatory responses elicited by the CpG DNA.     

吲哚美辛对乙酸诱导大鼠胃溃疡形成和愈合的影响

目的利用乙酸诱导大鼠胃溃疡模型研究吲哚美辛对化学诱导胃溃疡形成和愈合的影响,探讨其可能机制。方法雄性SD大鼠,体重160～180g。分两组,即单纯乙酸诱导胃溃疡作为对照组和乙酸诱导胃溃疡加吲哚美辛处理组,各时间点每组均8只。乙酸诱导胃溃疡后1、3和7d用RT-PCR和Westernblotting分别检测胃黏膜中环氧合酶(COX)和诱导型一氧化氮合酶(iNOS)的表达。用ELISA测定胃黏膜中PGE2量反映COX活性。同时研究吲哚美辛对iNOS表达、活性及胃黏膜损伤的影响,以溃疡面积来评估胃黏膜损伤程度。结果RT-PCR结果显示乙酸诱导大鼠胃溃疡后,COX2mRNA表达明显升高;以溃疡基底部为明显,3d最高,7d下降。胃黏膜PGE2合成也明显增高。吲哚美辛能抑制胃黏膜PGE2合成,处理组溃疡面积1d时为(52.6±6.1)mm2,小于对照组的(71.8±5.8)mm2(P＜0.05),且周围充血水肿较轻;3d时两组溃疡大小无差异,但吲哚美辛处理组溃疡基底部厚度为(11±0.5)mm,薄于对照组的(20±0.8)mm(P＜0.01);7d时吲哚美辛组溃疡而积为(35.4±3.5)mm2,大于对照组的(24.8±3.2)mm2(P＜0.05)。此外吲哚美辛能降低胃黏膜iNOS的表达及活性。结论吲哚美辛能减轻大鼠溃疡形成初期炎症反应,使组织免受进一步损伤,但使溃疡加深、延缓溃疡愈合。这一作用除和PGE2合成减少有关外,可能尚和抑制iNOS表达及活性有关。     

康莱特注射液对肺癌A549细胞环氧化酶作用的研究

目的:研究康莱特注射液对IL-1β刺激后A549肺癌细胞环氧化酶(COX)表达的抑制作用。方法:用RT-PCR方法检测100~300μL.mL^-1不同含量康莱特注射液作用前后A549肺癌细胞COX-1,COX-2 mRNA表达变化,同时用Western-blot检测COX-2蛋白表达变化。结果:A549细胞在康莱特注射液作用前后COX-1的mRNA表达无明显变化,但COX-2的mRNA和蛋白表达随着药物作用浓度增加而逐渐降低。结论:康莱特注射液能选择性抑制A549肺癌细胞COX-2表达。