重组人内抑素对大鼠佐剂性关节炎的影响

目的　观察重组人内抑素对大鼠佐剂性关节炎 (adju vantarthritis,AA)的影响及其作用机制。方法　用福氏完全佐剂 (CFA)诱导大鼠AA模型 ,MTT法检测脾淋巴细胞增殖反应 ,IL 1、IL 2活性的检测采用小鼠胸腺细胞增殖法 ,用放免法检测滑膜细胞培养上清液中IL 1和TNF α水平。结果　CFA致炎后d10 ,AA大鼠出现继发性炎症 ,给予不同剂量的内抑素 0 1、0 5、2 5mg·kg- 1·d- 1,sc ,连续 7d。结果发现 ,内抑素对AA大鼠的继发性足肿胀有抑制作用 ;进一步研究表明内抑素明显抑制AA大鼠过高的ConA诱导的脾细胞增殖反应 ,降低脾细胞IL 2的产生 ;对腹腔巨噬细胞(peritonealmacrophage ,PMΦ)产生过高的IL 1有抑制作用 ;另外 ,内抑素也可明显抑制AA大鼠滑膜细胞产生过高的IL 1和TNF水平。结论　重组人内抑素对AA大鼠具有治疗作用 ,其机制可能与其调节机体异常的免疫有关     

重组人内抑素对大鼠佐剂性关节炎继发性炎症的抑制作用

目的　观察重组人内抑素对大鼠佐剂性关节炎 (AA)继发性炎症是否有抑制作用。方法　用足容积测量仪检测足爪容积并对关节炎症程度进行目测评分 ;HE染色检测非致炎侧膝关节的病理改变。结果　福氏完全佐剂 (CFA)致炎后d 1 0 ,继发性炎症出现 ,同时给予不同剂量的内抑素(0 1、0 5、2 5mg·kg- 1 ·d- 1 ,sc) ,连续 7d。结果发现 ,内抑素 2 5mg·kg- 1 对AA大鼠的继发性足肿胀有明显的抑制作用 ;致炎后d 30病理检查显示AA大鼠的关节内滑膜增厚 ,滑膜衬层细胞增生并伴有新生血管形成 ,滑膜细胞有不同程度的脂肪变性 ;软骨及骨遭到破坏 ,且透明软骨内基质降解及新生血管形成。给予不同剂量的内抑素对AA大鼠关节组织的上述病理改变有不同程度的改善作用 ,内抑素 2 5mg·kg- 1 对AA大鼠的关节病理损伤有完全的抑制作用 ,同时可见滑膜组织内大量胶原沉积和滑膜细胞的脂肪变性。结论　重组人内抑素对大鼠佐剂性关节炎的继发性炎症有显著的抑制作用 ,并能阻止关节炎的病理改变     

重组人内抑素诱导佐剂性关节炎大鼠滑膜细胞凋亡的研究

目的以弗氏完全佐剂诱导的佐剂性关节炎(AA)大鼠为动物模型,观察重组人内抑素诱导AA大鼠滑膜细胞凋亡的作用。方法分离、培养滑膜细胞,噻唑蓝(MTT)比色法检测重组人内抑素体内用药对滑膜细胞增殖的影响;用缺口末端标记法(TUNEL)及流式细胞仪检测重组人内抑素对AA滑膜细胞凋亡的影响。结果AA大鼠表现为滑膜细胞异常增殖,rh-End(1.25、2.5、5.0mg·kg-1)体内治疗给药及rh-End6.25～50mg·L-1浓度范围体外给药可抑制AA大鼠成纤维样滑膜细胞的增殖反应。rh-End体内用药可诱导AA大鼠滑膜组织及细胞的凋亡,rh-End体外用药作用48h,AnnexinV/PI双染色法观察到大鼠AAFLS凋亡率为1.77%,rh-End(25mg·L-1)可诱导AA大鼠FLS的凋亡,其凋亡率为6.67%。结论rh-End可不同程度抑制FLS的增殖反应,并诱导其凋亡的发生,减轻AA大鼠增生性滑膜炎。上述结果提示炎症增生的滑膜细胞可能是rh-End的另一个作用靶点。     

重组人内抑素对佐剂性关节炎大鼠滑膜组织凋亡相关基因表达的影响

目的:观察重组人内抑素(rh-end)对佐剂性关节炎大鼠滑膜组织p53、fas和bcl-2基因表达的影响。方法:采用SYBR GreenⅠ实时荧光定量PCR方法,定量检测该药对佐剂性关节炎大鼠滑膜组织p53、fas和bcl-2 mRNA表达的影响。结果:与模型组相比,rh-end治疗组大鼠滑膜组织fas和bcl-2 mR- NA表达水平增加,p53 mRNA表达水平降低,差异有统计学意义(P<0.01)。结论:rh-end可经fas介导诱导佐剂性关节炎大鼠滑膜细胞凋亡,且不依赖于p53,不能被bcl-2所抑制。     

重组人内抑素对佐剂性关节炎大鼠滑膜细胞增殖及产生细胞因子的影响

目的观察重组人内抑素对体外培养佐剂性关节炎大鼠滑膜细胞的增殖功能及产生细胞因子的影响,探讨其治疗佐剂性关节炎的作用机制。方法采用弗氏完全佐剂(CFA)诱导的佐剂性关节炎(AA)大鼠模型,用足容积法测量关节肿胀度;采用collagenasetypeⅡ消化法分离、培养滑膜细胞,MTT法检测重组人内抑素体内用药对滑膜细胞增殖的影响;放免法检测滑膜细胞上清液中IL-1β、TNF-α的含量。结果CFA致炎后d10,AA大鼠出现继发性炎症,此时皮下注射重组人内抑素(1·25,2·5,5mg·kg-1),连续7d,对照组于d10给予MTX(1·0mg·kg-1)。各剂量组能明显抑制AA大鼠的继发性足肿胀;重组人内抑素体内用药可明显减少AA大鼠滑膜细胞数量,抑制滑膜细胞的增殖,并呈剂量依赖关系;各剂量组均可明显抑制AA大鼠滑膜细胞产生过高的IL-1β和TNF-α。结论重组人内抑素抑制AA大鼠滑膜细胞过度的增殖及过高的细胞因子是其治疗AA的途径之一。     

重组人内抑素对佐剂性关节炎大鼠滑膜组织血管内皮细胞生长因子表达的影响

目的:观察重组人内抑素对佐剂性关节炎大鼠滑膜组织血管内皮细胞生长因子(vascular endo-thelial cell growth factor,VEGF)的表达及其作用。方法:采用弗氏完全佐剂(Freund’s complete adjuvant,CFA)诱导的佐剂性关节炎(adjuvant-induced arthri-tis,AA)大鼠模型,用足容积法测量关节肿胀度;免疫组织化学法检测血管内皮细胞生长因子的表达及其对微血管密度的影响。结果:弗氏完全佐剂致炎后d 10,佐剂性关节炎大鼠出现继发性炎症,皮下注射给予不同剂量的重组人内抑素(1.25、2.5、5 mg.kg-1),连续7 d,对照组于d 10给予氨甲喋呤(Methotrexate,MTX,1.0 mg.kg-1)。重组人内抑素各剂量组能明显抑制AA大鼠的继发性足肿胀,重组人内抑素各剂量组可不同程度减少AA大鼠关节滑膜组织高表达的VEGF,还可使滑膜组织的微血管密度减少。结论:重组人内抑素可显著抑制AA大鼠滑膜组织VEGF的表达及微血管密度,该作用可能与抑制血管翳的生成及减少滑膜细胞表面VEGF的表达有关。     

重组人内抑素对佐剂性关节炎大鼠成纤维样滑膜细胞周期和PCNA表达的影响

目的观察重组人内抑素(rhEndostatin)对佐剂性关节炎(AA)大鼠成纤维样滑膜细胞(FLS)细胞周期及增殖细胞核抗原(PCNA)表达的影响,探讨rhEndostatin抑制AAFLS增殖的分子机制。方法制备AA大鼠模型,应用流式细胞仪分析rhEndostatin对AA FLS细胞周期的影响;分别采用实时荧光定量PCR和Western blot方法,定量分析rhEn-dostatin对AA大鼠滑膜组织PCNA mRNA及蛋白表达的影响。结果与正常组相比,AA FLS G1期细胞减少(P<0.01),S期和G2/M期细胞增加(P<0.01);AA大鼠滑膜组织PCNA mRNA和蛋白表达增加(P<0.05,P<0.01)。与AA模型组相比,rhEndostatin治疗组细胞G1期比例增加(P<0.01),S期和G2/M期细胞比例减少(P<0.01);滑膜组织PCNA mRNA和蛋白表达减少(P<0.01)。结论rhEn-dostatin可引起AA FLS细胞周期G1期阻滞,该作用可能与其抑制AA FLS PCNA表达有关。     

Anti-adjuvant arthritis of recombinant human endostatin in rats via inhibition of angiogenesis and proinflarnmatory factors

AIM: To investigate the profile of endostatin on adjuvant arthritis (AA) and angiogenesis blockade in synovitis. METHODS: The model of rat AA was induced by injection of intradermal complete Freund's adjuvant (CFA). Hind paw volume of rat was measured by volume meter and the activities of interleukin-1 (IL-1) and IL-2 were measured by the assay of thymocytes proliferation. IL-1β and tumor necrosis factor-a (TNF-α) produced     

Anti-adjuvant arthritis of recombinant human endostatin in rats via inhibition of angiogenesis and proinflammatory factors

AIM: To investigate the profile of endostatin on adjuvant arthritis (AA) and angiogenesis blockade in synovitis. METHODS: The model of rat AA was induced by injection of intradermal complete Freund's adjuvant (CFA). Hind paw volume of rat was measured by volume meter and the activities of interleukin-1 (IL-1) and IL-2 were measured by the assay of thymocytes proliferation. IL-1beta and tumor necrosis factor-alpha (TNF-alpha) produced by synoviocytes was estimated with radioimmunoassay. The number of new blood vessels in knee joint synovium was counted under microscope by hematoxylin and eosin (HE) staining. RESULTS: The secondary inflammation of AA rats appeared on the 10th day after injection of CFA. The therapeutic administration of endostatin (0.1, 0.5, and 2.5 mg/kg/d, sc, plus 7 d) was given from that time (d 10). It was found that endostatin significantly inhibited the secondary paw swelling and the number of new blood vessels in the synovium of AA rats. Endostatin significantly decreased the production of IL-1 derived from both peritoneal macrophages and synoviocytes and IL-2 from splenocytes, especially at the dose of 2.5 mg/kg. This effect of endostatin also was seen on TNF-alpha produced by synoviocytes. CONCLUSION: The recombinant human endostatin had an inhibitory effect on rat AA, which was related to its anti-angiogenesis and inhibition of proinflammatory cytokines.     

Relationship of blood markers to disease severity and drug efficacy in rat adjuvant arthritis.

Rat adjuvant arthritis (AA) was used as a model to evaluate several blood markers as possible predictive indicators of drug efficacy. AA was induced in Sprague-Dawley rats by the injection of complete Freund's adjuvant into the right hind foot pad. The rats were dosed p.o. from day 18 to day 31 with levamisole (10 mg/kg), indomethacin (1 mg/kg), diclofenac sodium (0.5 & 1 mg/kg), and prinomide (10 & 20 mg/kg). Disease severity was assessed by paw circumference on day 31. The following blood markers were analyzed: hyaluronate by ELISA, prostaglandin E2 by RIA, ESR by micro-dispette, total PMN by Technicon H-1, and albumin by BCG dye. Blood marker correlation (r) to disease severity was: hyaluronate (0.71), prostaglandin E2 (0.58), ESR (0.52), PMN (0.58), and albumin (-0.71). The relative rank order of drug efficacy (indomethacin, diclofenac sodium, and prinomide) did not differ using the change in paw circumference (day 31-day 17) or blood markers. Levamisole exacerbated the disease as measured by all the above parameters. Thus, these blood markers provide additional information for the statistical evaluation of drugs in rat adjuvant arthritis.     

盐诱导法高效表达重组人内抑素

内抑素是 1996年底由Ｆｏｌｋｍａｎ研究组首次发现的一种血管生成抑制剂 ,1997年 1月 ,Ｃｅｌｌ杂志报道 :内抑素特异性抑制内皮细胞增殖 ,并对血管生成与肿瘤生长具有很强抑制作用〔3〕。 1997年 11月 ,Ｎａｔｕｒｅ〔6〕 杂志又相继报道了Ｆｏｌｋ ｍａｎ研究组的内抑素研究的新成果 ,内抑素多次使用不引起小鼠副作用 ,说明这是一种反复用药后动物不产生耐药性的药物。由于内抑素具有广谱抗瘤效应、低毒性、可直接针对内皮细胞并且不产生药物耐受性 ,内抑素可能有着较大的临床应用前景。通过基因工程方法大量获取重组人内抑素将对该新型…     

人血管内皮抑素腺病毒注射液抑制脉络膜新生血管实验研究

目的探讨人血管内皮抑素腺病毒(Ad-Es)注射液对半导体激光诱导的BN大鼠脉络膜新生血管(CNV)的抑制作用。并比较不同给药方式的治疗效果,从而明确药物疗效及最佳给药途径,为进一步基因治疗临床试验奠定基础。方法雄性BN大鼠40只,右眼眼底采用半导体激光光凝方式建立CNV模型,随机分为4组,每组10只(10只眼)。光凝后21 d,A组于玻璃体腔内注射Ad-Es 0.01 mL;B组于球周注射Ad-Es 0.01 mL;C组于玻璃体腔内注射生理盐水0.01 mL;D组于球周注射生理盐水0.01 mL。观察给药后7 d各组的荧光眼底血管造影(FFA)荧光素渗漏情况,应用激光共焦显微镜下脉络膜血管平铺法测量各组CNV面积,光镜下观察组织细胞学变化,免疫组化检测CD105的表达。结果FFA晚期图像A组和B组视网膜光凝斑荧光素渗漏得到明显抑制,A组抑制效果更强;激光共焦显微镜下观察CNV面积,A组和B组CNV面积显著低于C、D组,A组CNV面积低于B组;组织病理学观察显示C、D组光凝区纤维血管较A、B组明显增多,A组低于B组;A、B组CNV相关表达因子CD105表达下降,A组较B组表达下降更明显。结论 Ad-Es注射液可以有效抑制动物模型的CNV表达,玻璃体腔注射给药抑制效果优于球周注射给药,为治疗CNV提供了一种新的可能途径。     

Determination of reactivity of resistance blood vessels in the isolated perfused legs of animals with inflammation as exemplified in adjuvant arthritis

Vasodepression was found ex vivo in the isolated perfused hind legs of rats, mice and guinea-pigs with paw inflammation using maximum pressure amplitude, EAm, pD2-value and intrinsic sensitivity (i.s.) as the test parameters of dose-response curves of vasopressor substances (noradrenaline, lys- and arg-vasopressin, angiotensin II, substance P, Na2-ATP). Vasodepression is strong in the anaphylactic and dextran paw edema, moderate in the carrageenin paw edema and adjuvant arthritis, but weak in the pertussis vaccine and kaolin paw edema. It is partly long-lasting, does not closely correlate with edema strength and can also be shown in the contralateral non-inflamed leg. Thus, a vasoreactivity depressing factor(s) must be liberated from the site of inflammation and reach the general circulation. Here, the method is described using the adjuvant arthritis as an example.     

木瓜苷通过抑制滑膜细胞功能减轻大鼠佐剂性关节炎

目的：观察木瓜苷(GCS)对大鼠佐剂性关节炎(AA)的作用,分析该作用与改善滑膜细胞的形态和功能之间的联系,方法：弗氏完全佐剂诱导大鼠AA,足容积法测量AA大鼠继发侧足容积,对关节疼痛和多发性关节炎进行评分,用11型胶原酶和胰蛋白酶消化和分离滑膜细胞,透射电镜进行滑膜细胞的超微结构观察,小鼠胸腺细胞增殖法测定滑膜细胞培养上清液中的IL—l水平,放射免疫法测定其中TNFα和PGE2含量．结果：GCS(60,120mg／kg)连续灌胃8天,可明显减轻大鼠AA的继发性足肿胀、炎性疼痛反应和多发性关节炎指数,改善滑膜细胞形态的异常并降低其高分泌的IL-1、TNFα和PGE2含量;GCS(120mg／kg)在减轻继发性炎症、疼痛和抑制滑膜细胞分泌PGE,方面明显优于阿克他利．结论：GCS具有减轻大鼠AA的作用,该作用的发挥可通过改善滑膜细胞异常的超微结构并抑制其过度分泌IL-1、TNFα和PGE2。     

Responses of human skin blood vessels to synthetic histamine analogues

1 The vascular responses of human skin to two synthetic analogues of histamine, 2-methyl histamine (an H₁-receptor agonist) and 4-methyl histamine (an H₂-receptor agonist) have been studied in vivo.

2 Both compounds evoked dose-related erythema, 2-methyl histamine but not 4-methyl histamine causing erythema mediated by an axon reflex, thus suggesting that the axon reflex and direct vasodilator action of histamine are due to H₁ and H₂ actions respectively.

3 The H₁-receptor antagonist chlorpheniramine inhibited erythema due to 2-methyl histamine. Cimetidine, an H₂-receptor antagonist, had no effect on the reaction to 2-methyl histamine. In contrast, the erythema reaction to 4-methyl histamine was suppressed by both cimetidine and chlorpheniramine.

4 Although both histamine analogues caused wealing, this was not dose-related within the dose range used, and neither chlorpheniramine nor cimetidine caused detectable suppression of wealing responses to either histamine analogue.

5 These results lend further support to the view that human skin blood vessels possess H₂ as well as H₁ receptors.

     

培美曲塞、顺铂联合重组人血管内皮抑制素治疗肺腺癌

目的观察培美曲塞、顺铂联合重组人血管内皮抑制素注射液（恩度）治疗肺腺癌的疗效与安全性。方法选择经病理及细胞学诊断明确的晚期肺腺癌初治患者52例,根据住院号随机分为观察组27例,对照组25例,观察组给予培美曲塞＋顺铂联合恩度治疗,对照组仅给予培美曲塞＋顺铂治疗,评价两组疗效及安全性。结果观察组客观有效率（48．1％）明显高于对照组（36．0％）（x2=7．82,P〈0．05）;观察组疾病控制率（92．6％）明显高于对照组（84．0％）（x2=4．05,P〈0．05）;观察组中位肿瘤进展时间（7．5个月）明显长于对照组（5个月）（t=5．71,P〈0．05）;两组在治疗期间均出现相似的不良反应,主要为白细胞减少、血红蛋白减少、血小板减少、恶心呕吐、便秘、心律失常、出血、乏力等,两组在不良反应分级情况差异均无统计学意义（均P〉0．05）。结论培美曲塞、顺铂联合恩度治疗肺腺癌疗效较单用化疗药疗效显著,且未增加不良反应,具有安全性,可作为肺腺癌的一线用药。     

Modulation of eicosanoid-induced contraction of mouse and rat blood vessels by gingerols

The effects of the active principles of crude ginger (a traditional Sino-Japanese medicine), the gingerols, on the contractile responses to eicosanoids were compared using isolated mouse and rat blood vessels. Leukotrienes (LT) C4 and D4, a thromboxane (TX) A2 derivative (U-46619), prostaglandins (PG) F2 alpha, PGI2-Na, PGE2, the stable PGI2 derivative TRK-100, and PGD2 induced contraction in longitudinal segments of mouse mesenteric veins in that order of potency. Exogenous arachidonic acid and PGE1 did not cause contraction. The mesenteric veins also contracted in response to noradrenaline (NA) and phenylephrine (PhE), but not to clonidine. The gingerols alone relaxed the muscle transiently and then augmented the response to PGF2 alpha, PGE2, PGI2-Na, and TRK-100, but suppressed the response to PGD2, U-46619, LTC4, LTD4, NA and PhE. (+/-)-[6]-Gingerol also potentiated the PGF2 alpha-induced contraction in longitudinal segments of rat mesenteric vein and vena cava, but inhibited it in circular segments of rat aorta and longitudinal segments of mouse mesenteric arteries. These results showed that (+/-)-[6]- and (+/-)-[8]-gingerols potentiated the contraction induced by prostanoids (except PGD2) and inhibited that produced by PGD2, TXA2, and LT, suggesting the modulation of eicosanoids-induced responses by (+/-)-[6]- or (+/-)-[8]-gingerol.