地塞米松脂质体壳聚糖胶囊对结肠炎大鼠肿瘤坏死因子的抑制作用

目的:评价地塞米松(DSP)脂质体冻干剂壳聚糖胶囊对结肠炎大鼠血清肿瘤坏死因子(TNF-α)活性的抑制作用.方法:用2,4,6-三硝基苯磺酸钠(TNBS)诱导大鼠结肠炎,建立反应结肠炎症程度的血清TNF-α和大鼠结肠重量与大鼠重量比值(C/B)的测定方法,观察口服各种DSP制剂对TNF-α的抑制程度.结果:灌肠后第5天时TNF-α达到最高值:(27.4±6.2)ng·L-1(n=10);各种DSP制剂均有降低TNBS诱导的实验性大鼠结肠炎血清TNF-α的作用,但DSP脂质体壳聚糖胶囊具有最好的抑制作用,各种剂量下血清TNF-α的值分别降低到(ng·L-1,n=10):30 μg:(18.9±3.3);60 μg:(12.2±3.0);90 μg:(8.6±2.2).结论:DSP脂质体冻干剂壳聚糖胶囊较DSP粉剂壳聚糖胶囊和其他DSP制剂在治疗结肠炎中可能具有更大的应用前景,值得进一步开发研究.     

4-氨基水杨酸口服结肠定位包衣片的体内药效学研究(英文)

目的：评价自制的4-氨基水杨酸口服结肠定位包衣片在大鼠体内的药效学。方法：以2,4,6-三硝基苯磺酸(TNBS)的乙醇溶液灌肠,制作溃疡性结肠炎大鼠模型。将大鼠随机分为7组：健康对照组,TNBS对照组,TNBS+低剂量(包衣片)组,TNBS+中剂量(包衣片)组,TNBS+高剂量(包衣片)组,TNBS+柳氮磺吡啶(SASP)组,TNBS+中剂量非包衣片组。大鼠连续给药5 d后,对结肠部位的大体形态学改变、组织学损伤以及组织髓过氧化物酶(MPO)活性进行评分或测定,评价4-氨基水杨酸(4-ASA)结肠定位包衣片在大鼠体内的药效学。结果：以TNBS灌肠可一次性制作大鼠溃疡性结肠炎模型。与4-ASA非包衣片组相比,包衣片组或SASP组显示,给药后动物结肠的大体形态改变评分、组织损伤评分及MPO活性均有所降低；高剂量组对各指标的影响与SASP无显著差异(P>0．05)。结论：与非包衣片比较,自制的4-氨基水杨酸口服结肠定位包衣片在治疗大鼠溃疡性结肠炎方面有更好的疗效。     

肠泻停胶囊对实验性结肠炎大鼠的影响

目的:探讨肠泻停胶囊对三硝基苯磺酸(TNBS)诱导大鼠实验性结肠炎的影响.方法:120只SD大鼠随机分为6组,每组20只:正常组、模型组、阳性对照组、肠泻停胶囊高、中、低组.除正常对照组未行造模外,其余五组大鼠均采用TNBS造模.肠泻停胶囊高、中、低组分别灌胃给药(1.80,0.90,0.45 g /kg体质量)、阳性对照组灌胃给予地塞米松剂量(0.2 mg/kg体质量)、其余组灌胃给予0.5%羧甲基纤维素钠溶液连续3 周、4周;观察肠泻停胶囊对大鼠腹泻率、死亡率、血白细胞计数、淋巴细胞百分率、脾脏及胸腺重量、组织形态学评分、组织MPO活性的影响.结果:经肠泻停胶囊干预后各剂量组动物腹泻明显缓解,腹泻率降低,动物死亡率降低,外周血WBC、LYM值及组织MPO值降低,剖检可见结肠组织溃疡面积明显缩小,水肿缓解,坏死减轻,未见肠壁增厚.结论:肠泻停胶囊连续给药,对实验性结肠炎治疗作用显著.     

HPLC同时测定乳癖消颗粒中三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1的含量

目的观察4-氨基水杨酸钠(4-ASANa)结肠靶向微丸对溃疡性结肠炎大鼠的干预作用及其机制。方法采用三硝基苯磺酸(TNBS)法制备大鼠实验性溃疡性结肠炎模型,通过4-ASANa结肠靶向微丸干预14d,观察大鼠结肠大体形态损伤、组织学变化和白细胞介素-1B(IL-1B)、肿瘤坏死因子-α(TNF-α)、超氧化物歧化酶(SOD)、丙二醛(MDA)、髓过氧化物酶(MPO)等指标的变化。结果4-ASANa结肠靶向微丸能减轻溃疡性结肠炎的病理损伤,显著降低溃疡性结肠炎大鼠血清IL-1B和TNF-a含量,差异具有统计学意义(P<0.05),降低结肠组织中MDA和MPO含量,而提高SOD活性,差异具有统计学意义(P<0.05),对化学因素引起的结肠炎有较好的治疗作用。结论4-ASANa结肠靶向微丸对TNBS诱导的大鼠结肠炎具有治疗作用,抑制IL-1B和TNF-a的增殖和表达,减轻自由基的损害可能是作用机制之一。     

同型半胱氨酸调控MEK-ERK-MLCK通路影响结肠炎大鼠肠黏膜通透性的实验研究

目的探讨同型半胱氨酸(homocysteine,Hcy)是否通过调控MEK-ERK-MLCK通路影响结肠炎大鼠肠黏膜通透性的机制。方法 SD大鼠分为4组,A组为正常对照组(NS皮下注射+NS灌肠),B组为正常对照+Hcy注射组(Hcy皮下注射+NS灌肠),C组为TNBS模型组(NS皮下注射+TNBS灌肠),D组为TNBS模型+Hcy注射组(Hcy皮下注射+TNBS灌肠)。建立高Hcy血症的实验性结肠炎大鼠模型,实验结束时取大鼠结肠组织病理学检查,并进行结肠匀浆检测MPO活性,采用Western blot方法检测大鼠小肠组织中MEK、ERK、p-ERK、MLCK、p-MLCK的蛋白表达水平,采用RT-q PCR方法检测大鼠小肠组织中MLCK mRNA表达。结果与正常对照组及模型对照组相比,TNBS模型+Hcy皮下注射组大鼠DAI及HI评分增高,结肠匀浆MPO活性增高,小肠黏膜组织MEK、ERK、p-ERK、MLCK、p-MLCK蛋白表达水平增加,MLCK mRNA相对表达量增加。结论 Hcy增加实验性结肠炎大鼠肠黏膜通透性,可能与调控MEK-ERKMLCK信号通路有关。     

4-氨基水杨酸钠结肠靶向微丸对溃疡性结肠炎大鼠免疫机制影响

目的 观察 4-氨基水杨酸钠(4-ASANa)结肠靶向微丸对溃疡性结肠炎大鼠的干预作用及其机制。方法 采用三硝基苯磺酸(TNBS)法制备大鼠实验性溃疡性结肠炎模型,通过4-ASANa结肠靶向微丸干预14 d,观察大鼠结肠大体形态损伤、组织学变化和白细胞介素-1B(IL-1B)、肿瘤坏死因子-α(TNF-α)、超氧化物歧化酶(SOD)、丙二醛(MDA)、髓过氧化物酶(MPO)等指标的变化。结果 4-ASANa结肠靶向微丸能减轻溃疡性结肠炎的病理损伤,显著降低溃疡性结肠炎大鼠血清IL-1B和TNF-a含量,差异具有统计学意义(P<0.05),降低结肠组织中MDA和MPO含量,而提高SOD活性,差异具有统计学意义(P<0.05),对化学因素引起的结肠炎有较好的治疗作用。结论 4-ASANa结肠靶向微丸对TNBS诱导的大鼠结肠炎具有治疗作用,抑制IL-1B和TNF-a的增殖和表达,减轻自由基的损害可能是作用机制之一。     

5-氨基水杨酸治疗结肠炎的抗氧化作用及对细胞因子表达的影响

目的探讨5-氨基水杨酸(5-ASA)灌肠治疗对三硝基苯磺酸(TNBS)诱导大鼠结肠炎的抗氧化作用及对细胞因子表达的影响。方法建立TNBS结肠炎模型,给予5-ASA灌肠治疗,同时设正常对照组和模型组。于治疗后1、2周,评价结肠大体、组织学损伤及髓过氧化物酶(MPO)活性,检测超氧化物歧化酶(SOD)活性、丙二醛(MDA)水平,逆转录-多聚酶链反应(RT-PCR)检测结肠组织白细胞介素(IL)-1β和肿瘤坏死因子(TNF)-αmRNA表达水平。结果5-ASA灌肠治疗能明显降低结肠炎大鼠的大体、组织学评分及MPO活性(P<0.05),升高SOD活性(P<0.05),降低MDA水平(P<0.05),降低IL-1β和TNF-α表达水平(P<0.05)。结论5-ASA灌肠治疗TNBS诱导的大鼠结肠炎疗效显著,其机制与抗氧化作用及抑制IL-1β和TNF-α的表达有关。     

具备高同型半胱氨酸血症的实验性结肠炎模型的建立

目的建立具备高同型半胱氨酸血症(HHcy)的实验性结肠炎模型,初步探讨Hcy对结肠炎症损伤的影响。方法将SD大鼠分为4组:正常对照组(NS灌肠+皮下注射NS)、正常对照+Hcy注射组(NS灌肠+皮下注射Hcy)、TN-BS模型对照组(TNBS/乙醇灌肠+皮下注射NS)、TNBS模型+Hcy注射组(TNBS/乙醇灌肠+皮下注射Hcy)。以TN-BS/乙醇混合溶液(100 mg·kg-1TNBS加入等体积NS)灌肠1次制备结肠炎模型后,采用皮下注射同型半胱氨酸(0.03μmol·kg-1,每天2次,间隔8 h,连续30 d)造成HHcy模型。实验结束后通过高效液相荧光法(HPLC-FD)检测血浆和结肠粘膜Hcy水平,进行DAI评分并取结肠组织进行HE染色评分,结肠组织匀浆检测MPO活性。结果与正常对照组比较,TNBS模型对照组大鼠DAI和HI评分增高(P<0.01),结肠组织MPO活性增高(P<0.01)。与TNBS模型对照组比较,TNBS模型+Hcy注射组大鼠血浆和结肠粘膜Hcy水平明显增高(P<0.01),DAI和HI评分明显增高(P<0.01),结肠组织MPO活性明显增高(P<0.01)。结论皮下注射同型半胱氨酸可以建立具备高同型半胱氨酸血症的实验性结肠炎模型,可用于探讨Hcy在结肠炎中的作用机制。     

芪茶益肠胶囊对大鼠化学性、免疫性结肠炎的影响

目的：探讨芪茶益肠胶囊对大鼠化学性、免疫性结肠炎的影响。方法：分别用醋酸和2，4，6-三硝基苯磺酸（TNBS）造大鼠化学性结肠炎和免疫性结肠炎模型，然后测定大鼠结肠黏膜组织形态和黏膜损伤指数并与柳氮磺吡啶比较。结果：芪茶益肠胶囊对减轻大鼠结肠黏膜急性炎症细胞浸润、抑制肉芽组织形成，降低损伤指数均有明显的作用。结论：芪茶益肠胶囊对实验动物溃疡性结肠炎有一定的治疗作用。     

甘草酸二铵抗大鼠溃疡性结肠炎作用及相关机制研究

目的:研究甘草酸二铵(DG)对2,4,6-三硝基苯磺酸(TNBS)诱导的大鼠溃疡性结肠炎的作用及可能作用途径。方法:以柳氮磺吡啶(SASP,225 mg.kg-1)为阳性对照,研究甘草酸二铵结肠给药(2,10,50 mg.kg-1,qd,共给药7 d)对TNBS性溃疡性结肠炎大鼠的结肠单位长度重量、结肠黏膜损伤评分、髓过氧化物酶(MPO)、结肠组织丙二醛(MDA)和超氧化物歧化酶(SOD)以及血清白介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)含量的影响;由HE染色、扫描和透射电镜观察结肠组织病理学变化;由免疫组化观察结肠组织环氧酶-2(COX-2)和细胞间黏附分子-1(ICAM-Ⅰ)表达的变化。结果:模型动物结肠组织发生明显病理学变化,伴有充血、溃疡以及大量的炎性细胞浸润,DG组均不同程度改善结肠组织病理学变化,降低单位长度结肠重量,并呈剂量依赖关系。与正常组比较,模型组大鼠结肠组织中MPO活性和MDA含量显著升高,SOD活性明显下降;DG能显著逆转模型组MPO,MDA和SOD的变化。与模型组比较,DG组结肠组织COX-2和ICAM-Ⅰ表达下调,血清中IL-1β和TNF-α表达量也有不同程度下降。结论:甘草酸二铵能有效改善溃疡性结肠炎大鼠的结肠炎症反应,其作用机制与抗氧化、降低促炎性细胞因子水平等有关。     

重组人粒细胞集落刺激因子壳聚糖胶囊在大鼠体内的吸收与药理活性

目的：评价重组人粒细胞集落刺激因子(rhG-CSF)壳聚糖胶囊经大鼠口服后的升白细胞作用和生物利用度。方法：于rhG-CSF冻干粉剂壳聚糖胶囊中加入不同量的胆酸钠(SC)后，在轻度乙醚麻醉下通过聚乙烯管给正常大鼠和环磷酰胺(CPA)所致白细胞降低大鼠口服胶囊，观察大鼠白细胞总数(BTL)经时变化规律。用氮兰四唑蓝(MTT)比色法测定各种rhG-CSF制剂的经时血药浓度，并计算曲线下面积和相对生物利用度。结果：口服rhG-CSF壳聚糖胶囊能升高大鼠BTL，同时对CPA诱发的大鼠BTL降低具有恢复作用，加入SC能产生协同作用。口服rhG-CSF壳聚糖胶囊的生物利用度分别为静脉注射的8．6％和皮下注射的17．8％。结论：口服rhG-CSF壳聚糖胶囊在升高白细胞作用中具有潜在的应用前景。     

Study on the colon specific delivery of prednisolone using chitosan capsules

In this study, we examined the effectiveness of chitosan capsules for the colon-specific delivery of prednisolone in rats. We also evaluated the effectiveness and side effects of prednisolone using chitosan capsules compared with the conventional dosage form (gelatin capsules). We found a significant increase in the concentration of prednisolone in the large intestinal mucosa when prednisolone was administered orally using chitosan capsules, as compared with the case using gelatin capsules. On the other hand, the plasma concentrations of prednisolone after oral administration using chitosan capsules were much lower than those in the case of gelatin capsules. We also assessed the effectiveness of prednisolone for the healing of trinitrobenzene sulfonic acid-induced colitis by measuring myeloperoxidase (MPO) activity and colon wet weight/body weight (C/B) ratio. MPO activities and C/B ratios were significantly reduced when prednisolone was administered orally using chitosan capsules, in comparison with the case of gelatin capsules. Moreover, the weight of the thymus, which is an index of the side effects of prednisolone, markedly decreased after oral administration of prednisolone using gelatin capsules, whereas its weight did not change as much when prednisolone was administered orally using chitosan capsules. These findings indicate that chitosan capsules might be useful for the colon-specific delivery of prednisolone and its enhanced effectiveness for the healing of colitis in rats. Moreover, chitosan capsules might be also effective in reducing the side effects of prednisolone due to its decreased intestinal transfer to the systemic circulation.     

葡萄籽原花青素对复发性结肠炎大鼠血清抗氧化能力及NO含量的影响

目的：观察葡萄籽原花青素（GSPE）对复发性结肠炎大鼠血清抗氧化能力及NO含量的影响,初步探讨葡萄籽原花青素治疗复发性结肠炎的作用机制。方法：直肠给予雄性Wistar大鼠80mg/kg2,4,6-三硝基苯磺酸（TNBS）/50%乙醇溶液复制结肠炎模型,在第16天时,用30mg/kgTNBS/50%乙醇溶液诱导结肠炎复发的模型。大鼠第二次致炎24h后,分别应用GSPE低、中、高剂量（100、200、400mg/kg）灌胃对其进行治疗,并以柳氮磺吡啶（SASP,500mg/kg）作为阳性对照。连续给药7d后处死所有大鼠,取结肠标本评价结肠湿重指数,生化法检测血清中髓过氧化物酶（MPO）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）和一氧化氮合酶（iNOS）活力及丙二醛（MDA）、谷胱甘肽（GSH）和NO含量。结果：与模型对照组比较,GSPE各剂量组大鼠体重下降程度较轻（P〈0.05或P〈0.01）,结肠湿重指数明显降低（P〈0.05或P〈0.01） 大鼠血清中MPO和iNOS活力及MDA和NO含量均明显降低（P〈0.05或P〈0.01） 大鼠血清中SOD和GSH-Px活力及GSH含量明显升高（P〈0.05或P〈0.01）。结论：GSPE可能通过提高复发性结肠炎大鼠血清抗氧化能力,抑制NO生成,来减轻复发性结肠炎炎症反应。     

龙胆苦苷对结肠炎小鼠的治疗作用

目的探究龙胆苦苷对结肠炎小鼠的治疗作用。方法采用2,4,6-三硝基苯磺酸(TNBS)-乙醇溶液诱导结肠炎小鼠模型,测定龙胆苦苷给药后对小鼠体质量、结肠指数、胸腺质量、结肠组织中髓过氧化物酶(MPO)活性以及血清炎症细胞因子TNF-α和IL-8的影响,并通过免疫组化法观察龙胆苦苷对小鼠结肠组织中环氧合酶-2(COX-2)蛋白表达的影响。结果给药1周后,龙胆苦苷各剂量组小鼠体质量与模型组相比显著改善;小鼠结肠指数、结肠组织中MPO活性以及炎性细胞因子TNF-α和IL-8与模型组相比均显著降低(P<0.05);免疫组化结果显示,龙胆苦苷能降低结肠炎小鼠组织中环氧合酶-2(COX-2)的蛋白表达。结论龙胆苦苷能修复TNBS-乙醇溶液诱导的小鼠结肠损伤,对小鼠结肠炎有显著的治疗作用。     

Effect of intracolonic benzalkonium chloride on trinitrobenzene sulphonic acid-induced colitis in the rat

Background:

We investigated the effects of benzalkonium chloride (BAC) on trinitrobenzene sulphonic acid (TNBS)-induced colitis in rats.

Methods:

TNBS was administered intrarectally before and/or after BAC treatment. In the first study, the effects of treatment with BAC 6, 12 or 24 h after TNBS were examined. In the second study, animals were treated with BAC before, after or before and after TNBS, and were examined 7 days later. The severity of colitis was assessed by macroscopic and histological scoring of the colonic damage and by determination of colonic myeloperoxidase (MPO) activity. Macrophages and CD4⁺ and CD8⁺ T cells were examined by immuno-histochemistry.

Results:

When BAC was instilled into the colon 6, 12 or 24 h after TNBS, weight loss and macroscopic and histological features of the colon were similar to that of controls (TNBS alone). In contrast, MPO activity was significantly reduced in all three groups post-treated with BAC. In the groups examined 7 days after TNBS treatment, rats post-treated with BAC exhibited increased weight gain and significantly reduced macroscopic damage and MPO activity compared to the TNBS control group. Rats pre-treated with BAC exhibited less macroscopic damage of the colon than rats receiving only TNBS, but histological damage, MPO and weight gain were unchanged from TNBS controls. Immunohistochemistry revealed that BAC pre-treatment increased the numbers of macrophages and T cells in the colon. After TNBS treatment, macrophage accumulation was evident in the colon, but T cells were scarce. However, these cells were preserved or enhanced in the colonic mucosa in TNBS-treated rats that had been pre-treated with BAC.

Conclusions:

Treatment with BAC, particularly after induction of colitis, produces a significant reduction in the severity of tissue injury and inflammation through mechanisms that are not fully understood.

     

黄芩汤抗 TNBS诱导的小鼠溃疡性结肠炎活性研究

目的：研究黄芩汤水煎液中主要成分及对2,4,6－三硝基苯磺酸钠（ TNBS）造模balb／c小鼠的溃疡性结肠炎的作用。方法 HPLC－MS法分析水煎液成分。雄性balb／c小鼠随机分为空白对照组,TNBS模型组,柳氮磺嘧啶阳性对照组（500 mg · kg－1）,黄芩汤水煎液1,2,4 g· kg－1治疗组。 TNBS造模7 d后,研究各组小鼠的结肠病理评分、微观评分、体质量、小鼠结肠长度和血清髓过氧化物酶（ MPO）。结果与标准品对照,黄芩汤水煎液中含有复方中主要成分,黄芩汤水煎液可使TNBS造模balb／c小鼠的体质量明显增加,改善身体的一般状况和小鼠结肠黏膜的损伤,下调MPO。结论黄芩汤水煎液中含有各药物的主要成分,对TNBS诱导的小鼠溃疡性结肠炎具有较好的治疗作用。     

PARP inhibition reduces acute colonic inflammation in rats

Poly(ADP-ribose) polymerases (PARP) comprise a family of enzymes which catalyse poly(ADP-ribosyl)ation of DNA-binding proteins. Multiple researches indicate the importance of PARP in promoting cell recruitment and thereby inducing organ injury in various forms of inflammation, such as colitis. We have evaluated the effects of two PARP inhibitors, nicotinamide and 1,5-dihydroxyisoquinoline, in acute colitis induced by trinitrobenzensulfonic acid (TNBS) in rats. Nicotinamide (20-40 mg/kg) and 1,5-dihydroxyisoquinoline (4-8 mg/kg) were administered 48, 24 and 1 h prior to the induction of colitis as well as 24 h later. 48 h after colitis induction the lesions were blindly scored and quantified as ulcer index. Histological study and colonic inflammation were assessed by gross appearance and myeloperoxidase (MPO) activity. Prostaglandin E2 (PGE2) synthesis and, cyclooxygenase-1 and cyclooxygenase-2 expressions by Western blotting and immunohistochemistry were also performed. Inflammation following TNBS induction was characterized by increased colonic wall thickness, oedema, diffuse inflammatory cells infiltration in the mucosa and necrosis. Furthermore, increased MPO activity, cyclooxygenase-2 expression and PGE2 synthesis were significantly augmented after TNBS instillation. On the contrary, treatment with 1,5-dihydroxyisoquinoline significantly reduced the degree of colon injury and also caused a substantial reduction in the rise in MPO activity, in the increase of staining for cyclooxygenase-2, as well as in the up-regulation of PGE2 caused by TNBS in the colon. Although nicotinamide significantly did not reduce macroscopic damage, it decreased both MPO activity and PGE2 colonic levels. In conclusion, we demonstrated that PARP inhibition can exert beneficial effects in experimental colitis and may, therefore, be useful in the treatment of ulcerative colitis.