Necrostatin-1对雨蛙肽诱导的小鼠急性胰腺炎的保护作用

目的观察Necrostatin-1(Nec-1)对雨蛙肽诱导的小鼠急性胰腺炎(AP)的影响。方法 C57BL/6小鼠予以50μg/kg的雨蛙肽腹腔注射7次,间隔1 h,建立AP模型。于第1次雨蛙肽注射后3 h、6 h、9 h和12 h,取出胰腺组织,HE染色。Western blot检测RIP1和RIP3蛋白的表达;C57BL/6小鼠随机分为:Control组、Vehicle组、Nec-1组和Nec-1i组,予以50μg/kg的雨蛙肽腹腔注射10次,间隔1 h,第1次雨蛙肽注射2 h前,腹腔注射Nec-1(1 mg/kg,每6 h 1次)、Nec-1i或等体积溶剂和空白对照。于第1次雨蛙肽注射后12 h、18 h和24 h,收集血清和胰腺组织。检测血清淀粉酶和脂肪酶。HE染色评估胰腺损伤程度。Real-time RT-PCR检测胰腺组织IL-1β和TNF-αmRNA的表达。结果 RIP1和RIP3蛋白在雨蛙肽诱导的AP中逐渐升高,且与胰腺腺泡细胞的坏死呈正相关;应用Nec-1后,与Vehicle组和Nec-1i组比较,于12 h、18 h和24 h,小鼠血清淀粉酶和脂肪酶水平明显降低,胰腺组织病理评分也明显减少,同时胰腺组织中IL-1β和TNF-αmRNA的水平也明显降低。结论 Nec-1对实验性胰腺炎具有明显的保护作用;程序性坏死可能促进了急性胰腺炎的损伤。     

不同剂量槲皮素对高甘油三酯血症相关性急性胰腺炎大鼠胰腺病理的影响

目的:观察不同剂量的槲皮素对高甘油三酯血症(hypertriglyceridemia,HTG)相关性急性胰腺炎(acute pancreatitis,AP)大鼠胰腺病理的影响.方法:72只♂SD大鼠,随机分为12组:HTG组(n=6,高脂饮食2 wk:77%普通饲料+3%胆固醇+20%猪油)、HTG+AP组(n=6,高脂饮食2 wk,腹腔内注射雨蛙肽50冚g/kg×2次,间隔1 h)、HTG+AP+槲皮素组(n=24,高脂饮食2 wk后再分为4组,诱发AP后分别给予50、100、150、200 mg/kg槲皮素腹腔注射)、正常血脂组(n=6,正常饮食2 wk)、正常血脂+AP组(n=6,正常饮食2 wk,腹腔内注射雨蛙肽50冚g/kg×2次,间隔1 h)、正常血脂+AP+槲皮素组(n=24,正常饮食2 wk后再分为4组,诱发AP后分别给予50、100、150、200 mg/kg槲皮素腹腔注射).所有大鼠均于AP造模后9 h处死,测定血浆淀粉酶活性,评价胰腺病理变化.结果:大鼠高脂饮食2 wk后,血清甘油三酯和总胆固醇均较正常饮食组明显升高,且差异有统计学意义(P0.001).与正常血脂+AP组相比,HTG+AP组的血浆淀粉酶活性(23670.00 U/L±2053.13 U/L vs 13136.00 U/L±3536.95 U/L)和胰腺病理学评分(9.75分±0.94分vs 5.92分±1.32分)均明显升高,且差异有统计学意义(P=0.022;P0.001).正常血脂+AP+槲皮素组和HTG+AP+槲皮素组的血浆淀粉酶活性均呈剂量依赖性地下降,胰腺病理损伤也相应地减轻,槲皮素的这种保护效应对HTG+AP组尤为明显(HTG+AP100、150、200 mg/kg槲皮素干预组vs未给药组总评分:均P0.001;正常血脂+AP 100、150、200 mg/kg槲皮素干预组vs未给药组总评分:P=0.084,P=0.003,P0.001).其中,又以腺泡坏死和炎性浸润改善得最为显著(HTG+AP 100、150、200 mg/kg槲皮素干预组vs未给药组坏死评分:均P0.001;炎性浸润评分:P=0.008,P=0.006,P=0.001).结论:槲皮素可以减轻大鼠HTG相关性AP的胰腺病理损伤,尤其是腺泡坏死和炎性浸润,且这一保护作用呈现剂量依赖性.     

JAM-C单抗对小鼠急性坏死性胰腺炎的抑制作用

目的:观察JAM-C单克隆抗体对小鼠急性坏死性胰腺炎(ANP)模型胰腺和全身炎症的抑制作用.方法:采用雨蛙素和脂多糖联合腹腔注射的方法建立小鼠重症急性胰腺炎模型.生理盐水对照组(NS组):小鼠给予腹腔注射无菌生理盐水(10 mL/kg),共注射6次,间隔1 h;ANP模型组(ANP组):小鼠给予腹腔注射雨蛙素(50μg...     

肝纤维化能够保护小鼠抵抗D-GalN/LPS诱导的致死性损伤

目的:证明四氯化碳(carbon tetrachloride,CCl4)诱导的肝纤维化小鼠对致死性D-氨基半乳糖/脂多糖(D-galactosamine and lipopolysaccharide,D-GalN/LPS)攻击的耐受性.方法:建立CCl4诱导的肝纤维化小鼠模型,于纤维化6 wk时以致死剂量的D-GalN(700 mg/kg)/LPS(50μg/kg)进行攻击,以同样处理的正常小鼠作为对照,即实验共分为4组:正常对照组(Nor)、急性损伤组(Nor+D-GalN/LPS)、肝纤维化组(Fib)、肝纤维化+急性攻击组(Fib+D-GalN/LPS).根据攻击前后小鼠生存率、转氨酶水平及肝组织学的变化来评估正常和纤维化小鼠对致死性D-GalN/LPS损伤的耐受性.结果:生存分析显示,Fib+D-GalN/LPS组的生存率显著高于Nor+D-GalN/LPS组(100%vs20%).血清转氨酶结果表明,Fib+D-GalN/LPS组肝损伤程度明显轻于Nor+D-GalN/LPS组,其sALT水平分别为(6630 U/L±1675 U/L)和(22429 U/L±5446 U/L)(P<0.01).接受攻击的纤维化和正常小鼠的sALT分别升高了14.3倍和455.9倍.肝组织学检查结果也证明,接受致死性D-GalN/LPS攻击的纤维化小鼠的肝损伤较同样处理的正常小鼠明显减轻.结论:CCl4诱导的肝纤维化可保护小鼠抵抗致死性D-GalN/LPS损伤的攻击.     

急性胰腺炎胰腺腺泡细胞不同死亡方式与细胞内酶释放的关系

目的 观察轻重程度不同的体外急性胰腺炎(AP)胰腺腺泡细胞发生凋亡、胀亡的状况及细胞内酶的释放情况,探讨两者的关系。方法 以两步酶消化法分离胰腺腺泡细胞,分为4组。AP组加入雨蛙素0.1μg/ml,细菌脂多糖(LPS)组加入雨蛙素及LPS( 10 mg/L),奥曲肽(OCT)组加入雨蛙素及OCT(100ng/ml),对照组加培养液。应用丫碇橙(AO)和溴乙锭(EB)双染色法检测腺泡细胞的凋亡及胀亡,采用比色法检测培养上清中淀粉酶及乳酸脱氢酶( LDH)含量。结果 对照组、AP组、LPS组、OCT组的细胞凋亡指数分别为2.2±0.4、6.4±0.6、4.6±0.4、11.2±1.2;胀亡指数分别为3.0±0.4、17.2±1.6、23.0±2.2、12.8±1.4;LDH的分泌量分别为(2180 ±240)、(8060±930)、(9460±920)、(6860±740) U/dl;淀粉酶的分泌量分别为(1750±190)、(3820±460)、(4420±480)、(2260±260) U/L。AP组、LPS组、OCT组的上述4项指标均显著高于对照组(P值均＜0.05);LPS组的胀亡指数及LDH和淀粉酶分泌量均较AP组显著增加(P值均＜0.05),而凋亡指数则较AP组显著减少(P＜0.05);OCT组的凋亡指数较AP组显著增多(P＜0.05),而胀亡指数及LDH、淀粉酶分泌均较AP组显著减少(P值均＜0.05)。结论 诱导AP胰腺腺泡细胞凋亡,并减少细胞胀亡的发生,可减少腺泡细胞内酶的释放。     

米诺环素对黑质急性炎症的保护作用及与自噬相关性研究

目的观察米诺环素对脂多糖(Lipopolysaccharide,LPS)模型小鼠黑质急性炎症的保护作用,并探讨其作用机制。方法选取3月龄C57BL雄性小鼠,设立对照组、LPS组、中剂量米诺环素组(50 mg M组)、高剂量米诺环素组(75 mg M组)及75 mg M+LPS组、50 mg M+LPS组、25 mg M+LPS组,其中LPS组为单次腹腔注射LPS(5 mg/kg)。随后进行爬杆试验的行为学检测;并采用免疫组化法观察黑质部位小胶质细胞数量,免疫印迹法检测小鼠黑质部位α-突触核蛋白(α-synuclein)、LC3-Ⅱ、P62和HDAC6蛋白表达量。结果与LPS组相比,75mg M+LPS组小鼠爬杆速度下降更显著(P0. 05),25 mg M+LPS组及50 mg M+LPS组小鼠爬杆速度则较LPS组明显加快(P0. 01); 25 mg M+LPS组及50 mg M+LPS组小鼠黑质部位小胶质细胞的活化数量减少,且α-synuclein、LC3-Ⅱ、p62、HDAC6蛋白水平下降(P0. 01),而75 mg M+LPS组α-synuclein、LC3-Ⅱ、p62、HDAC6蛋白水平有所增加(P0. 05)。结论低、中剂量米诺环素不仅能减轻LPS单次腹腔注射造成的小鼠黑质部位急性炎症反应,而且能缓解该部位出现的自噬功能障碍,减少p62和α-synuclein黑质部位的积聚,改善小鼠的运动能力。     

吡咯列酮对雨蛙肽诱导急性胰腺炎氧化应激过程的作用

目的 探讨过氧化物酶体增殖物激活受体(PPAR)γ激动剂吡咯列酮在雨蛙肽诱导大鼠急性胰腺炎中对氧化应激产物的影响及保护作用.方法 30只雄性SD大鼠随机分为对照组、雨蛙肽+不同剂量吡咯列酮组、雨蛙肽组、雨蛙肽+吡咯列酮+GW9662组.每组6只.急性胰腺炎造模30 min后处死大鼠,光镜下观察胰腺组织病理学变化,测定各组大鼠胰腺组织质量与体重比,比色法检测胰腺组织髓过氧化物酶(MPO)活性、丙二醛(MDA)和一氧化氮合酶(NOS)及组织诱导型一氧化氮合酶(iNOS)含量.结果 与对照组比较,雨蛙肽组胰腺组织水肿严重胰腺净重/体重(0.0072比0.0042)],MPO活性、MDA、NOS及iNOS含量升高(P＜0.01).与雨蛙肽组比较,吡咯列酮20 mg/kg及40 mg/kg组胰腺损伤减轻,胰腺净重/体重、MPO活性、MDA和NOS及iNOS含量降低(P＜0.05);与吡咯列酮40 mg/kg组比较,PPARγ拮抗剂GW9662逆转了吡咯列酮的保护作用(P＜0.05).结论 在雨蛙肽诱导的大鼠急性胰腺炎发病中,胰腺腺泡细胞的氧化应激损伤起了重要的作用,PPARγ激动剂吡咯列酮预先干预,通过降低氧化应激过程,对雨蛙肽诱导的大鼠急性胰腺炎有一定的保护作用.     

JAM-C在小鼠急性坏死性胰腺炎模型不同组织中的表达

目的:研究连接黏附分子C(JAM-C)在小鼠急性坏死性胰腺炎(ANP)模型胰腺、肾脏和肺脏组织中的表达.方法:采用雨蛙素和脂多糖联合腹腔注射的方法建立小鼠ANP模型.模型组(ANP组)小鼠腹腔内注射雨蛙素(50 μg/kg),连续6次,每次间隔1 h,在末次注射雨蛙素后,即向小鼠腹腔内注射脂多糖(LPS)(10 mg/kg);对照组小鼠腹腔内注射等体积生理盐水.在末次注射3 h后,用眼球取血法收集血液,检测血清淀粉酶,并取胰腺、肾脏、肺脏组织,通过Western blot印迹杂交法检测JAM-C在这些组织上的表达.结果:JAM-C在ANP组的胰腺、肺脏和肾脏上均有着高表达,大于生理盐水对照组3倍以上(0.608±0.133 vs 0.176±0.024,0.718± 0.148 vs 0.160±0.027,0.654±0.085 vs 0.166±0.039,均P<0.05).结论:在小鼠ANP模型中,JAM-C在胰腺、肾脏和肺脏上的表达均明显增高,提示JAM-C在ANP发病过程中起到重要的作用.     

雨蛙肽引起的鼠急性坏死性胰腺炎:丙谷胺、Benzotript和胰泌素的保护作用

急性坏死性胰腺炎炎症和坏死过程的起动机制仍不清楚,治疗上除支持疗法外亦无其它有效方法.本文描述在用雨蛙肽引起大鼠急性坏死性胰腺炎后的生化和组织结构改变的消退过程,并比较胰泌素与两种CCK受体拮抗剂,丙谷胺和benzotriPt的保护作用.材料和方法:实验用大鼠15只禁食16～18小时后,每小时接受不同剂量的雨蛙肽(5～200μg/kg)皮下或腹腔内注射共7次,对照组注射0.9%氯化钠,于首次注射后9小时全部杀死.在研究胰腺炎时间过程和药物保护作用的实验中,用雨蛙肽50μg/kg腹腔内注射共7次.杀死鼠的时间同前.每次给予雨蛙肽前半小时,皮下注射不同剂量的丙谷胺或benzotript或胰泌素.另作下述实验:1.首次雨蛙肽注射后12小时杀死动物,观察丙谷胺或胰泌素的作用.2.研究在第三次注射雨蛙肽后给予丙谷胺或benzotript 4次的作用.3.研究丙谷胺或胰泌素对小剂量雨蛙肽引起的胰腺炎的作用.对部分鼠进行较长时间的实验观察.结果:皮下或腹腔内注射大剂量雨蛙肽可引起血清     

鸢尾素对脂多糖诱导的小鼠炎性认知障碍的保护作用

目的 探讨鸢尾素(Irisin)对脂多糖(LPS)诱导的炎性认知障碍的作用及可能机制。方法 利用LPS腹腔注射C57BL/6J雄性小鼠建立炎性认知障碍小鼠模型。48只小鼠随机分为正常对照(Control)组、LPS组、LPS+多奈哌齐(Donepezil)组和LPS+Irisin组，每组12只。LPS+Donepezil组和LPS+Irisin组在LPS腹腔注射2 h后分别予小鼠灌胃3 mg/(kg·d)Donepezil、腹腔注射2 mg/(kg·d)Irisin, Control组腹腔注射相同体积的生理盐水。采用水迷宫评估各组认知功能；采用实时荧光定量聚合酶链反应(qRT-PCR)检测脑内白细胞介素(IL)-1β、IL-6和肿瘤坏死因子(TNF)-α mRNA相对表达水平；采用免疫荧光法观察Irisin对脑皮层和海马各区星形胶质细胞的激活水平；采用Western印迹检测各组磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)信号通路关键蛋白表达水平。结果 水迷宫结果：Irisin可使LPS模型小鼠2、3、4、5 d逃避潜伏期明显缩短，而目标象限停留时间、穿越原平台次数均显著增加(P&...     

The immunoneuroendocrine role of melatonin

Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.     

Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland

Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.     

Conservative management of perforated duodenal diverticulum： A case report and review of the literature

Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.     

Response of rat pineal melatonin to calcium, magnesium, and lithium is circadian stage dependent

Abstract: Herein we documented the response of pineal melatonin production to electrolytes known to be effective on pineal function in view of a possible circadian stage dependence. We studied the release of melatonin by perifused rat pineal glands at 2 different circadian stages corresponding to the middle of the light and dark periods, i.e., respectively, 7 and 19 HALO (Hours After Light Onset, L:D = 12:12). The initial efflux rates were, as expected, much higher in the perifusates of glands removed from rats sacrificed during the dark phase than of those removed during the light phase. After 3 hr of perifusion, melatonin release reached similar levels which were found constant up to the 8th hr of perifusion, whatever the circadian stage. Perifusion of the glands with physiological concentrations for the rat of calcium (5.2 mmol/1) and magnesium (1.34 mmol/1) resulted in a stimulatory effect on the pineal glands removed from rats sacrificed in the middle of the dark period (19 HALO), whereas no effects were observed on the pineal glands removed from rats sacrificed during the light (7 HALO). Lithium (0.28 and 0.55 mmol/1) was ineffective on melatonin release in pineal glands removed 7 and 19 HALO. Our results show differences in the initial efflux rates of melatonin and in the response of perifused pineal glands to calcium and magnesium according to the circadian stage.     

Presence of immunoreactive growth hormone and prolactin in the ovine pineal gland

Abstract: The use of antisera raised against bovine growth hormone (GH) and ovine prolactin (PRL) enabled the detection of related immunoreactive (ir) sequences of proteins in ovine pineal tissue. The isolation of PRL-like ir-material was accomplished using a 0.25 M ammonium sulphate (pH 5.5) extraction followed by ethanol precipitation, whereas the resulting 2.0 M ammonium sulphate (pH 7.0) precipitate contained a GH-like immunoreactivity. Gel chromatography of the GH-like immunoreactivity (Sephadex G-100) indicated the presence of several GH-like fragments ranging in the M_r range of 7,000 to 55,000. Analyses of the PRL-like ir-material found in pineal tissue on HPLC using a TSK 545-DEAE column led to the resolution into a single peak of immunoreactivity. A single peak of activity was also observed following chromatofocusing and hydrophobic interaction chromatography of the ir-peak from the TSK 545-DEAE column. The PRL-like ir-material inhibited the binding of [¹²⁵I]ovine PRL-S14 to anti-ovine PRL antibodies without showing an affinity for binding to anti-rat PRL or anti-bovine GH antibodies. Scatchard analysis of the binding of pineal PRL-like ir-material and pituitary ovine PRL-S14 to liver membranes from day-20 pregnant rats revealed similar affinity constants (K_a of 4.7 ± 0.2 × 10⁹ M^-1). In addition, the replication of Nb 2 Node rat lymphoma cells was stimulated by pineal PRL-like ir-material, an effect known to be specific for lactogenic hormones. The pineal PRL-like immunoreactivity appeared on sodium dodecyl sulfate polyacrylamide gels as a single major band of M_r 24,000. The functional status of PRL-and GH-like ir-material in the ovine pineal remains to be determined, but evidence is presented that the overall protein synthesis rate of the rat pineal responded to circulating concentrations of PRL.     

Microbiology of human immunodeficiency virus anorectal disease

PURPOSE: Individuals who are seropositive for the human immunodeficiency virus are at high risk for opportunistic infection and anorectal disorders. Little prospective information is available regarding anorectal pathogens in these patients. METHODS: One hundred sixty-three HIV-seropositive patients presented to the colorectal clinic between 1989 and 1992. Forty-seven (29 percent) patients were thought to have an infectious process and were prospectively studied using a standardized multiculture protocol. RESULTS: Mean age was 33 (range, 19–59) years. All were male; high-risk behavior accounted for 87 percent of HIV transmissions. Presenting complaints included anorectal pain (79 percent), pus per anum (28 percent), and blood per anum (26 percent). Examination revealed perianal tenderness (60 percent), condyloma (38 percent), perianal ulcers (38 percent), and anal fissures (34 percent). Sixty-six sets of cultures were performed; 28 patients had one set, 15 had two sets, and 4 had three sets. Thirty-two of these 47 patients (68 percent) had positive cultures including herpes (50 percent), cytomegalovirus (25 percent),Neisseria gonorrhoeae (16 percent), chlamydia (16 percent), acidfast bacilli (2 percent), and others (9 percent). Six of 32 patients with positive cultures had more than one organism cultured. Sixteen (50 percent) patients with positive cultures were treated medically, 8 (25 percent) were treated surgically and 8 (25 percent) were treated with both modalities. Sixty-one procedures were performed on 17 patients for condylomata. Eighteen patients had 20 procedures for abscesses, 50 percent of whom had positive cultures for other than common bowel flora; all improved. Fourteen patients underwent 33 procedures for perianal fistulas.Mycobacterium fortuitum was cultured from one patient who required 13 procedures for abscesses and fistulas. Forty-five (96 percent) patients were followed for an average of 12.5 months ±2.9 SEM (range, 1–94 months). Symptoms were improved or resolved in 22 of 32 (69 percent) patients with positive cultures and in 11 of 13 (84 percent) with negative cultures. CONCLUSIONS: Specific pathogens may often be identified in human immunodeficiency virus-seropositive patients with anorectal disorders if aggressively sought. Although patients without specific pathogens identified may be expected to improve with planned empiric treatment, positive identification allows more directed therapy.