13C-caffeine breath test identifies single nucleotide polymorphisms associated with caffeine metabolism

We performed a caffeine (N-3-methyl-¹³C) breath test (CafeBT) to determine whether it can be employed to identify caffeine metabolism-associated single nucleotide polymorphisms. The study included 130 healthy adults (mean age: 21.9 years). Saliva was collected using an Oragene®•DNA saliva collection kit. Breath samples were collected from the subjects. The subjects orally ingested 100 mg ¹³C-caffeine dissolved in distilled water. Subsequently, breath samples were collected in bags every 10 min for a total of 90 min. An analysis of ¹³CO₂ in the expired breath was performed by infrared spectroscopy, and the sum of Δ¹³CO₂ over 90 min (S^90m) was calculated. DNA from saliva samples was genotyped using TaqMan® SNP Genotyping for the following genes: cytochrome P4501A2: rs762551, rs2472297, aryl-hydrocarbon receptor (rs4410790), and adenosine A_2A receptor (rs5751876). All subjects had the genotype CC in rs2472297 alleles. No significant difference was observed in S^90m among the genotypes of rs762551 and rs5751876; however, a significant difference was found in S^90m among the genotypes of rs4410790 (C > T). Our findings suggest that the N-3 demethylation of caffeine is dependent on the rs4410790 allele and that CafeBT may be used to determine rs4410790 genotypes.     

枸橼酸咖啡因与氨茶碱对早产儿神经行为发育影响分析

目的 比较枸橼酸咖啡因和氨茶碱对早产儿神经行为发育的影响,为早产儿早期合理干预提供临床依据。方法 选择2014年6月-2018年6月在中国西电集团医院新生儿科住院接受枸橼酸咖啡因治疗的原发性呼吸暂停(AOP)早产儿62例作为研究组;2011 年 12 月-2014年5月同在中国西电集团医院新生儿科住院采用氨茶碱治疗的AOP患儿69例作为对照组。出生3~8 d及矫正胎龄40周时分别行头颅MRI检查,评估脑白质损伤(WMD)并进行两组比较;6个月及12个月时应用Gesell婴幼儿发育量表测试比较两组神经行为发育水平。结果 两组患儿在性别、出生胎龄及体重、产前孕母糖皮质激素应用及分娩方式、5 min Apgar评分、辅助通气和表面活性物质的应用等方面差异均无统计学意义(P>0.05)。首次头颅MRI显示两组患儿WMD差异无统计学意义(P>0.05)。矫正胎龄40周时头颅MRI显示,咖啡因组WMD较氨茶碱组明显改善,差异有统计学意义(P<0.05);且6个月及12个月时Gessell婴幼儿发育量表测试,随访至校正6月龄时,咖啡因组患儿的大运动、精细运动及个人-社交评分均高于氨茶碱组(P<0.05);随访至校正12月龄时,咖啡因组患儿的大运动、精细运动、语言、适应性评分均高于氨茶碱组(P<0.05)。结论 枸橼酸咖啡可明显改善AOP患儿6个月及12个月时的神经行为发育。     

枸橼酸咖啡因对新生大鼠缺氧缺血性脑损伤后神经细胞增生与凋亡及长期学习能力的影响

目的研究枸橼酸咖啡因(CC)对新生大鼠缺氧缺血性脑损伤(HIBD)后神经细胞增生与凋亡及长期学习记忆能力的影响。方法 7日龄SD新生大鼠随机分为假手术组、HIBD组、CC组,每组16只;HIBD组及CC组经左颈总动脉结扎并缺氧制作HIBD模型,CC组在HI前、HI后0 min、24 h、48 h、72 h给予CC 20 mg/kg腹腔注射,假手术组和HIBD组分别在同一时间点给予等量生理盐水腹腔注射;同时从生后10日龄起腹腔注射5-溴脱氧尿嘧啶核苷(Brd U)标记新生细胞,剂量50 mg/kg,每12 h 1次,共5次。12日龄时每组随机选取8只大鼠处死,免疫组织化学染色检测海马齿状回颗粒下层5-溴脱氧尿嘧啶核苷(Brd U)和海马CA1区活化半胱氨酸天冬氨酸蛋白酶(活化Caspase-3)的表达情况,TUNEL法测定海马CA1区神经细胞凋亡情况,其余各组大鼠28日龄时行Y迷宫学习和记忆能力测试。结果三组新生大鼠脑组织中均可见Brd U阳性细胞,差异有统计学意义(F=101.38,P0.01);HIBD组、CC组Brd U阳性细胞数均较假手术组增多,差异有统计学意义(P0.05)。三组新生大鼠海马CA1区均可见活化Caspase-3阳性细胞,差异有统计学意义(F=379.77,P0.01);CC组活化Caspase-3阳性细胞较HIBD组显著减少,但仍多于假手术组,差异有统计学意义(P0.05)。三组新生大鼠海马CA1区中均可见TUNEL阳性细胞,差异有统计学意义(F=505.92,P0.01),以HIBD组最多,假手术组最少,两两比较差异均有统计学意义(P0.05)。Y迷宫实验结果,各组大鼠达标所需训练总次数的差异有统计学意义(F=32.05,P0.01),以HIBD组所需训练次数最多。24 h后正确反应率在三组间的差异也有统计学意义(F=24.99,P0.01),以HIBD组大鼠正确反应率最低。结论枸橼酸咖啡因可以改善HIBD大鼠长期学习记忆力,其机制可能与通过减少缺氧缺血后神经细胞的凋亡有关。     

The effect of caffeine abstinence on sleep among habitual caffeine users with poor sleep

Caffeine is the most widely used psychoactive substance in the world and is known to disrupt healthy sleep. However, very few studies have directly tested the effect of caffeine abstinence on sleep, and these have yielded inconsistent findings. The purpose of the present study was to examine changes in sleep following caffeine abstinence and examine the extent to which characteristics of habitual caffeine use moderated this change. Participants included 66 healthy, young adults with habitual caffeine use and poor sleep. During the 2‐week baseline, sleep was assessed using wrist actigraphy and daily caffeine use was assessed with bedtime diaries. Eligible participants then completed 1 week of caffeine abstinence, during which sleep was measured with wrist actigraphy. Multilevel models found no significant differences between either mean levels or growth trajectories of total sleep time or sleep efficiency between baseline and caffeine abstinence. Mean levels of sleep onset latency also did not differ between baseline and caffeine abstinence. A small but significant quadratic effect was observed, such that sleep onset latency decreased during the first few days of caffeine abstinence, then increased to levels above baseline. Characteristics of caffeine use did not moderate changes in sleep between baseline and caffeine abstinence. These data suggest that abstaining from caffeine may not result in long‐term sleep improvement for habitual caffeine users, which contradicts the common sleep health recommendation. The present findings encourage more rigorous investigation of the effectiveness of caffeine restriction on sleep.     

Early Renal Injury Induced by Caffeine Consumption in Obese,Diabetic ZSF1 Rats

Our previous studies indicate that prolonged caffeine consumption exacerbates renal failure in nephropathy associated with the metabolic syndrome. Reduced activity of the antioxidant defense system and beneficial effects of antioxidant therapy have been reported in diabetic rats and humans. The purpose of this study was to examine the early renal effects of caffeine consumption and the effects of concomitant antioxidant therapy in young obese, diabetic ZSF1 rats. Eleven-week-old male ZSF1 rats were randomized to drink tap water, caffeine (0.1%), tempol (1 mmol/L), or a solution containing caffeine and tempol for nine weeks. Caffeine significantly reduced body weight and glycosuria (weeks 2–9), improved glucose tolerance (week 9), had no effect on elevated plasma triglycerides, plasma cholesterol (week 9) and blood pressure (week 9), and significantly increased plasma cholesterol level (weeks 5 and 9). Yet, as early as after two weeks, caffeine greatly augmented proteinuria and increased renal vascular resistance (RVR) and heart rate (HR: week 9). Tempol had no effects on metabolic status and development of proteinuria, did not alter caffeine-induced metabolic changes and early proteinuria, and attenuated caffeine-induced increase in HR and RVR. Immunohistochemical analysis revealed significant glomerular and interstitial inflammation, proliferation, and fibrosis in control animals. Caffeine augmented the influx of glomerular and interstitial macrophages (ED1+ cells) influx, glomerular and tubular proliferative response, and glomerular collagen IV content. Tempol abolished the exacerbation of renal inflammation, proliferation, and fibrosis induced by caffeine. In conclusion, in nephropathy associated with the metabolic syndrome, caffeine—most likely through the interaction with adenosine receptors and interference with anti-inflammatory and/or glomerular hemodynamic effects of adenosine—augments proteinuria and stimulates some of the key proliferative mechanisms involved in glomerular remodeling and sclerosis. Tempol does not prevent early renal injury (i.e., proteinuria) induced by caffeine, yet abolishes late renal inflammatory, proliferative, and fibrotic change induced by chronic caffeine consumption in obese ZSF1 rats.     

咖啡因对辐射诱导细胞凋亡和G2／M期阻滞的影响

用形态学,DNA电泳和流式细胞仪等方法观察γ射线诱发BAF3细胞G_2/M期阻滞和凋亡的作用,以及咖啡因对其的影响。结果显示,5Gyγ射线照射后6小时引起细胞G_2/M期阻滞,G_2/M期比例为49.10％,12小时升至52.00％,无细胞凋亡发生。照前分别加入1mmol/L及5mmol/L咖啡因,照后12小时G_2/M阻滞程度降低,比例降至39.42％和15.49％,形态学检测细胞凋亡比例为8.33±1.53％和18.33±1.76％,流式细胞仪检测细胞凋亡比例为18.78％和43.81％,DNA电泳可见细胞凋亡特有的梯状图谱。结论提示,G_2/M期阻滞抑制能促进辐射诱导的细胞凋亡。     

Protecting effect of caffeine against vinblastine (an anticancer drug) induced genotoxicity in mice

Vinblastine a DNA non-intercalating agent has wide application against several human neoplasms, and found to cause cytogenotoxicity. In this study, clastogenotoxicity of vinblastine (1.5?mg/kg b w) and its prevention by caffeine at different doses (25, 50 and 100?mg/kg b w) administered intraperitoneally was assessed in in vivo mice. It was found that micronucleus level had decreased significantly (up to 28.8%) in 100?mg caffeine treated group at 30?h post treatment. However, it did not exhibit protective effect against chromosomal aberration in spaermatogonial cells at 24?h post treatment. The frequencies of aberrant primary spermatocytes had decreased significantly in 25 and 100?mg caffeine at 4th week of post treatment. Similarly, in 100?mg of caffeine administered, abnormal sperm level had reduced (4.01%) significantly at 8th week post treatment. Thus, caffeine decreased the vinblastine induced chromosomal aberrations and mitotic index in bone marrow cells. In conclusion, this study shows that caffeine exerts protective effect against vinblastin induced cytogenotoxicity. Further studies on molecular mechanism are interesting in order to develop it as an effective drug in cancer chemotherapy.