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Kynurenic Acid Levels and Kynurenine Aminotransferase I, II and III Activities in Ganglia, Heart and Liver of Snail Helix Pomatia. 螺螺神经节、心脏和肝脏犬尿氨酸水平及犬尿氨酸转氨酶I、II和III活性
Q3 CELL BIOLOGY Pub Date : 2023-08-18 DOI: 10.33594/000000643
Carina Kronsteiner, Halina Baran, Berthold Kepplinger
<p><strong>Background/aims: </strong>Kynurenic acid (KYNA), a tryptophan metabolite along the kynurenine pathway, is an endogenous antagonist of glutamate ionotropic excitatory amino acid (EAA) receptors and the α7 nicotinic acetylcholine receptor (nAChR). The involvement of KYNA in various pathological conditions and during the aging process is significant. KYNA synthesis from L-kynurenine (L-KYN), through the action of several kynurenine aminotransferases (KATs), is present in the central nervous system (CNS) and periphery of mammals. We were interested in investigating the ability of the brain and peripheral organs of Helix pomatia snails to synthesize KYNA, in an in vitro study. In comparative studies between rat and snail, we looked for the synthesis of KYNA in the liver. We then looked for an effect of age on KYNA synthesis.</p><p><strong>Methods: </strong>Ten shell parameters of the Helix pomatia snail were used to establish an Age Rating Scale (ARS), i.e. body weight, shell weight, shell length, width and height, shell opening length and width, lip width, number of shell turns and external shell growth rings. An age of the snails was determined according to the ARS and the snails were divided into three groups, i.e. young, middle and old age. Homogenates of dissected regions, i.e. cerebral ganglia (CG), subpharyngeal ganglia (SG) consisting of pedal, visceral and pleural ganglia, heart and liver, were examined. KYNA was measured by high performance liquid chromatography (HPLC) and KAT activities were measured by an enzymatic method.</p><p><strong>Results: </strong>With respect to ARS, an evaluation of the age of the snails between young (1-2 years), middle (5-7 years) and old (9-13 years) showed significant differences (p<0.001). Analysis of KYNA levels in different snail tissues, i.e. CG, SG, heart and liver, showed an occurrence in the low femtomolar range. Marked and significant increases of KYNA were found in the liver of middle and old age groups. In the SG, KYNA decreased significantly with age. There were no differences in KYNA levels between groups in CG and heart. The lowest KAT activity was found in CG and SG (5 pmol/mg/h), while in heart and liver the values were visibly higher (between 8 and 80 pmol/mg/h). Only in the liver, and exceptionally only for KAT I, the activity increased significantly with age, i.e. up to 14 years. No age-related changes in KAT I, II and III activities were found in CG and SG. Snail liver shows a different pattern of KAT activities compared to the rat liver.</p><p><strong>Conclusion: </strong>Regions of the CNS and periphery of the snail Helix pomatia are able to synthesize KYNA due to KAT activities. In the snail liver, KAT I activity increased with age. Notably, there was no age-related increase in KAT activities in the heart and especially in the CNS of Helix pomatia, indicating significant differences from mammals. A moderate KYNA metabolism in the Helix pomatia snail in the periods studied, up t
背景/目的:犬尿氨酸(KYNA)是一种内源性谷氨酸嗜离子性兴奋性氨基酸(EAA)受体和α7烟碱乙酰胆碱受体(nAChR)的拮抗剂,是沿犬尿氨酸途径的色氨酸代谢物。KYNA参与各种病理条件和衰老过程是重要的。l -犬尿氨酸(L-KYN)通过多种犬尿氨酸转氨酶(KATs)的作用合成KYNA,存在于哺乳动物的中枢神经系统(CNS)和外周。在体外研究中,我们感兴趣的是研究螺旋螺的大脑和外周器官合成KYNA的能力。在大鼠和蜗牛的比较研究中,我们寻找肝脏中KYNA的合成。然后我们寻找年龄对KYNA合成的影响。方法:采用螺壳质量、壳重、壳长、壳宽、壳高、壳开长、壳开宽、壳唇宽、壳转数、壳外生长环等10个参数,建立年龄评定量表(ARS)。根据ARS测定钉螺年龄,并将钉螺分为青年、中年和老年3组。解剖区域的匀浆,即大脑神经节(CG),咽下神经节(SG)组成的踏板,内脏和胸膜神经节,心脏和肝脏,进行了检查。采用高效液相色谱法测定KYNA,酶法测定KAT活性。结果:在ARS方面,幼龄(1 ~ 2岁)、中年(5 ~ 7岁)和老年(9 ~ 13岁)钉螺的年龄评价存在显著差异(p)。结论:由于KAT活性的存在,中枢神经系统区域和钉螺尾部螺旋外围能够合成KYNA。在蜗牛肝脏中,KAT I活性随着年龄的增长而增加。值得注意的是,在心脏中,特别是在螺旋鱼的中枢神经系统中,KAT活性没有与年龄相关的增加,这表明与哺乳动物有显著差异。在长达14年的研究期间,螺旋螺的适度KYNA代谢可能是一种生理现象,可以保护器官免受因高KYNA水平而可能出现的功能不足,正如所建议的那样。因此,寻找能够调节蜗牛体内KYNA浓度的因子是合理的。
{"title":"Kynurenic Acid Levels and Kynurenine Aminotransferase I, II and III Activities in Ganglia, Heart and Liver of Snail Helix Pomatia.","authors":"Carina Kronsteiner,&nbsp;Halina Baran,&nbsp;Berthold Kepplinger","doi":"10.33594/000000643","DOIUrl":"https://doi.org/10.33594/000000643","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Background/aims: &lt;/strong&gt;Kynurenic acid (KYNA), a tryptophan metabolite along the kynurenine pathway, is an endogenous antagonist of glutamate ionotropic excitatory amino acid (EAA) receptors and the α7 nicotinic acetylcholine receptor (nAChR). The involvement of KYNA in various pathological conditions and during the aging process is significant. KYNA synthesis from L-kynurenine (L-KYN), through the action of several kynurenine aminotransferases (KATs), is present in the central nervous system (CNS) and periphery of mammals. We were interested in investigating the ability of the brain and peripheral organs of Helix pomatia snails to synthesize KYNA, in an in vitro study. In comparative studies between rat and snail, we looked for the synthesis of KYNA in the liver. We then looked for an effect of age on KYNA synthesis.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;Ten shell parameters of the Helix pomatia snail were used to establish an Age Rating Scale (ARS), i.e. body weight, shell weight, shell length, width and height, shell opening length and width, lip width, number of shell turns and external shell growth rings. An age of the snails was determined according to the ARS and the snails were divided into three groups, i.e. young, middle and old age. Homogenates of dissected regions, i.e. cerebral ganglia (CG), subpharyngeal ganglia (SG) consisting of pedal, visceral and pleural ganglia, heart and liver, were examined. KYNA was measured by high performance liquid chromatography (HPLC) and KAT activities were measured by an enzymatic method.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;With respect to ARS, an evaluation of the age of the snails between young (1-2 years), middle (5-7 years) and old (9-13 years) showed significant differences (p&lt;0.001). Analysis of KYNA levels in different snail tissues, i.e. CG, SG, heart and liver, showed an occurrence in the low femtomolar range. Marked and significant increases of KYNA were found in the liver of middle and old age groups. In the SG, KYNA decreased significantly with age. There were no differences in KYNA levels between groups in CG and heart. The lowest KAT activity was found in CG and SG (5 pmol/mg/h), while in heart and liver the values were visibly higher (between 8 and 80 pmol/mg/h). Only in the liver, and exceptionally only for KAT I, the activity increased significantly with age, i.e. up to 14 years. No age-related changes in KAT I, II and III activities were found in CG and SG. Snail liver shows a different pattern of KAT activities compared to the rat liver.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusion: &lt;/strong&gt;Regions of the CNS and periphery of the snail Helix pomatia are able to synthesize KYNA due to KAT activities. In the snail liver, KAT I activity increased with age. Notably, there was no age-related increase in KAT activities in the heart and especially in the CNS of Helix pomatia, indicating significant differences from mammals. A moderate KYNA metabolism in the Helix pomatia snail in the periods studied, up t","PeriodicalId":9845,"journal":{"name":"Cellular Physiology and Biochemistry","volume":"57 4","pages":"279-297"},"PeriodicalIF":0.0,"publicationDate":"2023-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10088933","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A High-Fat Diet Induces Cardiac Damage in Obesity-Resistant Rodents with Reduction in Metabolic Health. 高脂肪饮食诱导抗肥胖啮齿动物心脏损伤,降低代谢健康。
Q3 CELL BIOLOGY Pub Date : 2023-08-16 DOI: 10.33594/000000642
Janete Corrêa Cardoso, Vinicius Valois Pereira Martins, Amanda Rangel Madureira, Suellem Torezani Sales, Filipe Martinuzo Filetti, Camila Renata Corrêa, Breno Valentim Nogueira, Ana Paula Lima-Leopoldo, André Soares Leopoldo

Background/aims: Obesity resistance is associated with the complex interaction of stringent and environmental factors that confer the ability to resist mass gain and body fat deposition, even when eating high-calorie diets. Considering that there are numerous gaps in the literature on the metabolic processes that explain Obesity resistance, specifically in relation to oxidative stress, the purpose of the study was to investigate whether obesity-resistant (OR) rats develop elevated reactive oxygen species in cardiac tissue.

Methods: Wistar rats were initially randomized into two groups: a standard diet (SD) and a high-fat diet (HFD) group. The SD and HFD groups were further divided into control (C), OR, and obese prone (OP) subgroups based on body weight. This criterion consisted of organizing the animals in each group in ascending order according to body weight (BW), and the cutoff point was identified in the animals by terciles: 1) lower BW; 2) intermediate BW; and 3) higher BW. Rats were sacrificed on the 14th week, and serum and organs were collected. Nutritional assessment, food profiles, histological analysis, comorbidities, and cardiovascular characteristics were determined.

Results: BW showed a significant difference between the standard diet and high-fat diet groups in the 4th week of the experimental protocol, characterizing obesity. In the 4th week, after the characterization of Obesity resistance, there was a significant difference in BW between groups C, OP, and OR. The OP and OR groups showed a significant increase in caloric intake in relation to the C group. The OP group showed a significant increase in final BW, retroperitoneal fat pad mass, sum of corporal fat deposits and reactive oxygen species, in relation to groups C and OR. The area under the glycemic curve, insulin resistance index and basal glucose were elevated in the OP group in relation to the C. OP also promoted an increase in HOMA-IR when compared with C. OR rats showed a non-significant increase in insulin and HOMA-IR in OR vs. C (p = ~0.1), but no significant differences were observed between OP vs. OR for these parameters, suggesting that both groups suffered from decreased metabolic health. Total cardiac mass, left ventricular cross-sectional area, and cholesterol levels were significantly elevated in the OP and OR groups compared with the C group.

Conclusion: A high-fat diet induces cardiac damage in obesity-resistant rodents with reduction in metabolic health.

背景/目的:肥胖抵抗与严格和环境因素的复杂相互作用有关,这些因素赋予了抵抗体重增加和身体脂肪沉积的能力,即使在食用高热量饮食时也是如此。考虑到在解释肥胖抵抗的代谢过程,特别是与氧化应激有关的文献中存在许多空白,本研究的目的是研究肥胖抵抗(Obesity -resistant, OR)大鼠是否会在心脏组织中产生较高的活性氧。方法:将Wistar大鼠随机分为标准饮食组(SD)和高脂饮食组(HFD)。SD组和HFD组根据体重进一步分为对照(C)、OR和肥胖倾向(OP)亚组。该标准是按体重(BW)由高到低依次组织各组动物,并按以下顺序确定分界点:1)体重较低;2)中间体重;3)更高的体重。第14周处死大鼠,采集血清和脏器。确定了营养评估、食物概况、组织学分析、合并症和心血管特征。结果:在实验方案的第4周,标准饮食组和高脂肪饮食组的体重出现了显著差异,具有肥胖特征。第4周,在完成肥胖症抵抗特征后,C组、OP组和OR组的体重差异有统计学意义。与C组相比,OP组和OR组的热量摄入显著增加。与C组和OR组相比,OP组的最终体重、腹膜后脂肪垫质量、体脂肪沉积总量和活性氧含量均显著增加。与C相比,OP组的血糖曲线下面积、胰岛素抵抗指数和基础葡萄糖均升高。与C相比,OP组也促进了HOMA-IR的升高(p = ~0.1), OR大鼠的胰岛素和HOMA-IR与C相比无显著升高(p = ~0.1),但OP组与OR组在这些参数上没有显著差异,表明两组的代谢健康都有所下降。与C组相比,OP组和OR组的心脏总质量、左室横截面积和胆固醇水平均显著升高。结论:高脂肪饮食可引起抗肥胖啮齿动物心脏损伤,降低代谢健康。
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引用次数: 1
The Absence of an Na+/Ca2+Exchanger (NCX) in Bullfrog Proximal Tubules and Cellular pH is More Influential Than Cellular Ca2+ on Proximal Na Transport. 牛蛙近端小管和细胞pH中Na+/Ca2+交换剂(NCX)的缺失对近端Na转运的影响大于细胞Ca2+。
Q3 CELL BIOLOGY Pub Date : 2023-07-31 DOI: 10.33594/000000641
Yutaka Matsumura

Background/aims: The functional significance of the Na+/Ca2+ exchanger (NCX) in basolateral membranes in the proximal tubule remains controversial. The key factor in crosstalk between the apical and basolateral sides is not known.

Methods: We investigated the basolateral membranes, using double-barreled Ca2+ or pH ion-selective microelectrodes. We used doubly perfused bullfrog kidneys in vivo, and switched the basolateral solution (renal portal vein) to experimental solutions.

Results: In the control, cellular pH (pHi) was 7.33 ± 0.032 (mean ± SE, n = 7) and in separate experiments, cellular Ca2+ activity (aCai) was 249.6 ± 35.54 nM (n = 28). Changing to respiratory acidosis, pHi was significantly acidified by 0.123 pH units on average and the change of aCai was +53.1 nM (n = 9 ns). In metabolic acidosis, pHi was reduced by 0.151 while aCai was reduced by 143.4. Using the 30 mM K+ solution, pHi was increased by 0.233 while aCai was reduced by 203.9, with depolarization. Both ionomycin and ouabain caused aCai to increase. In the 0.5 mM Na+ solution (replaced with BIDAC Cl), pHi was reduced by 0.177. No changes in aCai (+49.8 ns) were observed although we recorded depolarization of 15.2 mV. In the 0.5 mM Na+ solution, replaced with raffinose, no changes in aCai (-126.4 ns) were observed with depolarization (6.5 ns).

Conclusion: Our results suggest that thermodynamic calculations of cellular Na+ concentration led to the conclusion that either a Na+/HCO3- exchanger (NBC) or NCX could be present in the same basolateral membrane. H+ ions are the most plausible key factor in the crosstalk.

背景/目的:Na+/Ca2+交换器(NCX)在近端小管基底外侧膜中的功能意义仍然存在争议。在根尖侧和基底侧之间产生串扰的关键因素尚不清楚。方法:采用双管Ca2+或pH离子选择性微电极对基底外侧膜进行研究。我们在体内使用双灌注牛蛙肾脏,并将基底外侧溶液(肾门静脉)转换为实验溶液。结果:对照组细胞pH (pHi)为7.33±0.032 (mean±SE, n = 7),单独实验细胞Ca2+活性(aCai)为249.6±35.54 nM (n = 28)。pHi转变为呼吸性酸中毒,平均显著酸化0.123 pH单位,aCai变化+53.1 nM (n = 9 ns)。代谢性酸中毒时,pHi降低0.151,aCai降低143.4。在30 mM K+溶液中,pHi增加了0.233,aCai减少了203.9。离子霉素和沃卡因均引起aCai升高。在0.5 mM Na+溶液中(用BIDAC Cl代替),pHi降低了0.177。虽然我们记录了15.2 mV的去极化,但aCai (+49.8 ns)没有变化。在0.5 mM Na+溶液中,用棉子糖代替,去极化(6.5 ns)时aCai (-126.4 ns)没有变化。结论:我们的研究结果表明,细胞内Na+浓度的热力学计算得出结论,Na+/HCO3-交换剂(NBC)或NCX可能存在于同一基底外侧膜中。氢离子是串扰中最可信的关键因素。
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引用次数: 0
De Novo Cloning and Functional Characterization of a Mechanosensitive Piezo-Like Ion Channel in the Crayfish. 小龙虾机械敏感压电离子通道的克隆与功能表征。
Q3 CELL BIOLOGY Pub Date : 2023-07-29 DOI: 10.33594/000000640
Bora Ergin, Berk Saglam, Kaan Arslan, Nazlı Coskun Beyatli, Zihni Ekim Taskiran, Turgut Bastug, Nuhan Purali

Background/aims: Mechanosensitive ion channels are the principal elements in the transduction of mechanical force to neural activity. To date, considerably fewer studies have been published about the molecular and structural properties of mechanosensitive channels. Piezo channels are the only ion channel family in eukaryotes which is selectively gated by the membrane tension. Piezo channels have been described in mammals and some other eukaryotes. However, not much information is available for the crustaceans.

Methods: Conventional cloning methods were used to clone the putative PIEZO channel mRNA in crayfish ganglia samples. HEK293T cells were transfected by the plasmid of the cloned gene for functional studies. The CDS of the mRNA translated into the protein sequence and three-dimensional structure of the channel has been calculated.

Results: An mRNA, 9378 bp, was firstly cloned from crayfish which codes a 2674 residues protein. The cloned sequence is similar to the piezo channel mRNAs reported in the other species. The sequence of the coded protein has been analyzed, and some functional domains have been identified. A three-dimensional structure of the coded protein was successfully calculated in reference to mouse piezo 1 channel protein data. A plasmid with a fluorescent protein indicator was synthesized for heterologous expression in HEK293T cells. The evoked calcium response to mechanical stimulation was not different from those observed in the control cells. However, the transfected cells were more sensitive to the gating modifier YODA-1.

Conclusion: Based on the apparent similarity in sequence, structure and functional properties to other known piezo channels, it has been proposed that cloned mRNA may code a piezo-like ion channel in crayfish.

背景/目的:机械敏感离子通道是机械力向神经活动传导的主要因素。迄今为止,发表的关于机械敏感通道的分子和结构特性的研究相当少。压电通道是真核生物中唯一一种由膜张力选择性门控的离子通道家族。在哺乳动物和其他一些真核生物中也发现了压电通道。然而,关于甲壳类动物的信息并不多。方法:采用常规克隆方法克隆小龙虾神经节中推定的PIEZO通道mRNA。用克隆基因质粒转染HEK293T细胞进行功能研究。计算了翻译成蛋白质的mRNA的CDS序列和通道的三维结构。结果:从小龙虾中首次克隆到全长9378 bp的mRNA,该mRNA编码2674个残基。克隆的序列与其他物种中报道的压电通道mrna相似。对编码蛋白的序列进行了分析,并确定了一些功能域。参考小鼠压电1通道蛋白数据,成功计算出编码蛋白的三维结构。合成了带荧光蛋白指示剂的质粒,并在HEK293T细胞中进行了异源表达。机械刺激引起的钙反应与对照细胞无明显差异。然而,转染后的细胞对门控修饰物YODA-1更敏感。结论:基于其序列、结构和功能特性与其他已知压电通道的相似性,克隆的mRNA可能在小龙虾中编码一个压电样离子通道。
{"title":"De Novo Cloning and Functional Characterization of a Mechanosensitive Piezo-Like Ion Channel in the Crayfish.","authors":"Bora Ergin,&nbsp;Berk Saglam,&nbsp;Kaan Arslan,&nbsp;Nazlı Coskun Beyatli,&nbsp;Zihni Ekim Taskiran,&nbsp;Turgut Bastug,&nbsp;Nuhan Purali","doi":"10.33594/000000640","DOIUrl":"https://doi.org/10.33594/000000640","url":null,"abstract":"<p><strong>Background/aims: </strong>Mechanosensitive ion channels are the principal elements in the transduction of mechanical force to neural activity. To date, considerably fewer studies have been published about the molecular and structural properties of mechanosensitive channels. Piezo channels are the only ion channel family in eukaryotes which is selectively gated by the membrane tension. Piezo channels have been described in mammals and some other eukaryotes. However, not much information is available for the crustaceans.</p><p><strong>Methods: </strong>Conventional cloning methods were used to clone the putative PIEZO channel mRNA in crayfish ganglia samples. HEK293T cells were transfected by the plasmid of the cloned gene for functional studies. The CDS of the mRNA translated into the protein sequence and three-dimensional structure of the channel has been calculated.</p><p><strong>Results: </strong>An mRNA, 9378 bp, was firstly cloned from crayfish which codes a 2674 residues protein. The cloned sequence is similar to the piezo channel mRNAs reported in the other species. The sequence of the coded protein has been analyzed, and some functional domains have been identified. A three-dimensional structure of the coded protein was successfully calculated in reference to mouse piezo 1 channel protein data. A plasmid with a fluorescent protein indicator was synthesized for heterologous expression in HEK293T cells. The evoked calcium response to mechanical stimulation was not different from those observed in the control cells. However, the transfected cells were more sensitive to the gating modifier YODA-1.</p><p><strong>Conclusion: </strong>Based on the apparent similarity in sequence, structure and functional properties to other known piezo channels, it has been proposed that cloned mRNA may code a piezo-like ion channel in crayfish.</p>","PeriodicalId":9845,"journal":{"name":"Cellular Physiology and Biochemistry","volume":"57 4","pages":"226-237"},"PeriodicalIF":0.0,"publicationDate":"2023-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9897309","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Autophagy Inhibition Increased Sensitivity of Pancreatic Cancer Cells to Carbon Ion Radiotherapy. 抑制自噬可提高胰腺癌细胞对碳离子放疗的敏感性
Q3 CELL BIOLOGY Pub Date : 2023-07-19 DOI: 10.33594/000000639
Makoto Sudo, Hiroko Tsutsui, Shuhei Hayashi, Koubun Yasuda, Keiko Mitani, Nana Iwami, Makoto Anzai, Toshiro Tsubouchi, Mitsuaki Ishida, Sohei Satoi, Tatsuaki Kanai, Seiko Hirono, Etsuro Hatano, Jiro Fujimoto

Background/aims: Pancreatic cancer has the poorest survival rate among all cancer types. Therefore, it is essential to develop an effective treatment strategy for this cancer.

Methods: We performed carbon ion radiotherapy (CIRT) in human pancreatic cancer cell lines and analyzed their survival, apoptosis, necrosis, and autophagy. To investigate the role of CIRT-induced autophagy, autophagy inhibitors were added to cells prior to CIRT. To evaluate tumor formation, we inoculated CIRT-treated murine pancreatic cancer cells on the flank of syngeneic mice and measured tumor weight. We immunohistochemically measured autophagy levels in surgical sections from patients with pancreatic cancer who received neoadjuvant chemotherapy (NAC) plus CIRT or NAC alone.

Results: CIRT reduced the survival fraction of pancreatic cancer cells and induced apoptotic and necrotic alterations, along with autophagy. Preincubation with an autophagy inhibitor accelerated cell death. Mice inoculated with control pancreatic cancer cells developed tumors, while those inoculated with CIRT/autophagy inhibitor-treated cells showed significant evasion. Surgical specimens of NAC-treated patients expressed autophagy comparable to control patients, while those in the NAC plus CIRT group expressed little autophagy and nuclear staining.

Conclusion: CIRT effectively killed the pancreatic cancer cells by inhibiting their autophagy-inducing abilities.

背景/目的:胰腺癌是所有癌症类型中生存率最低的。因此,开发一种有效的治疗策略对这种癌症至关重要。方法:对人胰腺癌细胞系进行碳离子放射治疗(CIRT),分析其存活、凋亡、坏死和自噬情况。为了研究CIRT诱导的自噬的作用,在CIRT之前向细胞中添加自噬抑制剂。为了评估肿瘤的形成,我们将cirt处理过的小鼠胰腺癌细胞接种在同基因小鼠的腹部,并测量肿瘤的重量。我们用免疫组织化学方法测量了接受新辅助化疗(NAC)加CIRT或单独接受NAC的胰腺癌患者手术切片中的自噬水平。结果:CIRT降低胰腺癌细胞的存活率,诱导凋亡和坏死改变,并伴有自噬。用自噬抑制剂预孵育加速细胞死亡。接种对照胰腺癌细胞的小鼠出现肿瘤,而接种CIRT/自噬抑制剂处理的细胞的小鼠出现明显的逃避。NAC治疗患者的手术标本表达与对照组相当,而NAC + CIRT组的手术标本表达很少自噬和核染色。结论:CIRT通过抑制胰腺癌细胞的自噬诱导能力有效杀伤胰腺癌细胞。
{"title":"Autophagy Inhibition Increased Sensitivity of Pancreatic Cancer Cells to Carbon Ion Radiotherapy.","authors":"Makoto Sudo, Hiroko Tsutsui, Shuhei Hayashi, Koubun Yasuda, Keiko Mitani, Nana Iwami, Makoto Anzai, Toshiro Tsubouchi, Mitsuaki Ishida, Sohei Satoi, Tatsuaki Kanai, Seiko Hirono, Etsuro Hatano, Jiro Fujimoto","doi":"10.33594/000000639","DOIUrl":"10.33594/000000639","url":null,"abstract":"<p><strong>Background/aims: </strong>Pancreatic cancer has the poorest survival rate among all cancer types. Therefore, it is essential to develop an effective treatment strategy for this cancer.</p><p><strong>Methods: </strong>We performed carbon ion radiotherapy (CIRT) in human pancreatic cancer cell lines and analyzed their survival, apoptosis, necrosis, and autophagy. To investigate the role of CIRT-induced autophagy, autophagy inhibitors were added to cells prior to CIRT. To evaluate tumor formation, we inoculated CIRT-treated murine pancreatic cancer cells on the flank of syngeneic mice and measured tumor weight. We immunohistochemically measured autophagy levels in surgical sections from patients with pancreatic cancer who received neoadjuvant chemotherapy (NAC) plus CIRT or NAC alone.</p><p><strong>Results: </strong>CIRT reduced the survival fraction of pancreatic cancer cells and induced apoptotic and necrotic alterations, along with autophagy. Preincubation with an autophagy inhibitor accelerated cell death. Mice inoculated with control pancreatic cancer cells developed tumors, while those inoculated with CIRT/autophagy inhibitor-treated cells showed significant evasion. Surgical specimens of NAC-treated patients expressed autophagy comparable to control patients, while those in the NAC plus CIRT group expressed little autophagy and nuclear staining.</p><p><strong>Conclusion: </strong>CIRT effectively killed the pancreatic cancer cells by inhibiting their autophagy-inducing abilities.</p>","PeriodicalId":9845,"journal":{"name":"Cellular Physiology and Biochemistry","volume":"57 4","pages":"212-225"},"PeriodicalIF":0.0,"publicationDate":"2023-07-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9841372","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Unprecedentedly High Level of Intracellular Vitamin C and DNA Epigenetic Marks in Prostate: Relevant for Male Fertility? 前列腺细胞内前所未有的高水平维生素C和DNA表观遗传标记:与男性生育能力有关?
Q3 CELL BIOLOGY Pub Date : 2023-06-19 DOI: 10.33594/000000638
Jolanta Guz, Ewelina Zarakowska, Pawel Mijewski, Aleksandra Wasilow, Justyna Szpotan, Marek Foksinski, Bartosz Brzoszczyk, Daniel Gackowski, Piotr Jarzemski, Ryszard Olinski

Background/aims: Seminal plasma composition is affected by the physiological state of the prostate, the major male reproductive gland. Semen components, like vitamin C, can modulate sperm function. Vitamin C is an effective scavenger of free radicals and is an essential component of enzymes such as TET proteins involved in the DNA demethylation process. In the present study, a broad range of parameters which may influence the metabolic state of the prostate gland were analysed including blood and prostate tissue vitamin C, epigenetic DNA modifications and 8-oxo-7,8-dihydro-2'-deoxyguanosine in DNA of leukocytes and prostate tissues.

Methods: The experimental material were tissue samples from patients with benign prostatic hyperplasia (BPH), normal/marginal prostate tissues from prostate cancer patients, leukocytes from healthy donors, and blood plasma from BPH patients and healthy donors. We applied ultra-performance liquid chromatography methods with mass spectrometry and/or UV detection.

Results: We found an unprecedentedly high level of intracellular vitamin C in all analysed prostatic tissues (benign prostatic hyperplasia and normal, marginal ones), a value much higher than in leukocytes and most human tissues. DNA epigenetic patterns in prostate cells are similar to other soft tissues like the colon, however, its uniqueness is the unprecedentedly high level of 5-(hydroxymethyl)-2'-deoxyuridine and a significant increase in 5-formyl-2'-deoxycytidine value compared to aforementioned tissues. Moreover, the level of 8-oxo-7,8-dihydro-2'-deoxyguanosine, an established marker of oxidative stress, is significantly higher in prostate tissues than in leukocytes and many previously studied soft tissues.

Conclusion: Our results pointed out that prostatic vitamin C (regarded as the main supplier of the vitamin C to seminal plasma) and the DNA modifications (which may be linked to the regeneration of prostate epithelium) may play important role to maintain the prostate health.

背景/目的:精液成分受男性主要生殖腺体前列腺的生理状态影响。精液成分,如维生素C,可以调节精子的功能。维生素C是一种有效的自由基清除剂,是参与DNA去甲基化过程的TET蛋白等酶的重要组成部分。在本研究中,分析了可能影响前列腺代谢状态的广泛参数,包括血液和前列腺组织维生素C,表观遗传DNA修饰和白细胞和前列腺组织DNA中的8-氧-7,8-二氢-2'-脱氧鸟苷。方法:实验材料为良性前列腺增生(BPH)患者的组织标本、前列腺癌患者的正常/边缘前列腺组织、健康供者的白细胞、BPH患者和健康供者的血浆。我们采用了超高效液相色谱法与质谱和/或紫外检测。结果:在所分析的前列腺组织(良性前列腺增生和正常边缘前列腺组织)中,我们发现细胞内维生素C水平空前高,远高于白细胞和大多数人体组织。前列腺细胞的DNA表观遗传模式与结肠等其他软组织相似,但其独特之处在于其5-(羟甲基)-2′-脱氧尿苷的水平前所未有地高,5-甲酰基-2′-脱氧胞苷的值与上述组织相比显著增加。此外,前列腺组织中的8-氧-7,8-二氢-2'-脱氧鸟苷水平明显高于白细胞和许多先前研究过的软组织。结论:前列腺维生素C(被认为是精浆中维生素C的主要供应者)和DNA修饰(可能与前列腺上皮的再生有关)可能在维持前列腺健康中起重要作用。
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引用次数: 2
Role of O-GlcNAcylation in Breast Cancer Biology. o - glcn酰化在乳腺癌生物学中的作用。
Q3 CELL BIOLOGY Pub Date : 2023-06-10 DOI: 10.33594/000000633
Karolina Kozal, Anna Krześlak

Breast cancer is the most common type of cancer in women. It has been extensively researched over the past decades, but the underlying mechanisms of its growth, proliferation, invasion, and metastasis require further investigation. Dysregulation of O-GlcNAcylation which is one of the most abundant post-translational modifications, impacts on the malignant features of breast cancer. O-GlcNAcylation is broadly recognized as a nutrient sensor and participates in cells' survival and death. Through its involvement in protein synthesis and energy metabolism, especially glucose metabolism, O-GlcNAcylation enables adaptation to a hostile environment. It supports the migration and invasion of cancer cells and may be crucial for breast cancer metastasis. This review summarizes the current state of knowledge about O-GlcNAcylation in breast cancer: the origins of its dysregulation, its effect on the different aspects of breast cancer biology, and the potential utility in diagnostics and therapy.

乳腺癌是女性中最常见的癌症类型。在过去的几十年里,人们对其进行了广泛的研究,但其生长、增殖、侵袭和转移的潜在机制需要进一步研究。o - glcnac酰化的失调是最丰富的翻译后修饰之一,影响着乳腺癌的恶性特征。o - glcnac酰化被广泛认为是一种营养传感器,参与细胞的生存和死亡。通过参与蛋白质合成和能量代谢,特别是葡萄糖代谢,o - glcn酰化使其能够适应恶劣的环境。它支持癌细胞的迁移和侵袭,可能是乳腺癌转移的关键。本文综述了o - glcn酰化在乳腺癌中的研究现状:其失调的起源,对乳腺癌生物学不同方面的影响,以及在诊断和治疗中的潜在应用。
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引用次数: 0
A Model of Interaction Between Apocynin and NADPH Oxidase Enzyme to Analyze the Possible Targets Responsible for Inhibition by Computational Analysis 一个罗布麻苷与NADPH氧化酶相互作用的模型,通过计算分析来分析可能的抑制目标
Q3 CELL BIOLOGY Pub Date : 2023-06-10 DOI: 10.33594/000000632
Vaibhav Gandhi, Ishan Wadi, T. Gupta, Divya Jindal, Pranav Pancham, Ashok Tiwari, S. Jha, R. Tiwari, Silpi Chanda, Chakresh Kumar, Jain, Manisha Singh
Background/Aims: A multi-component enzyme system called NADPH oxidase (NOX) helps innate immunity by generating reactive oxygen species (ROS). NOX hyperactivation has been associated w several diseases. This enzyme is a membrane-bound complex made up of six subunits when it is active. These enzymatic subunits are considered to be potent inhibitors of enzyme activity and good targets for reducing oxidative stress. Methods: The present study aimed to analyze the possible targets: the different subunits of NOX, for their interactions with apocynin to identify its possible mechanism of inhibition for NOX, using in silico tools. Monomer, dimer, and trimer of apocynin were docked to various subunits of NOX. Results: Comparable glide scores were obtained when the monomer and dimer of apocynin were docked with p47phox complete subunit of NOX and were better than in comparison to trimer. Free Energy of Binding (FEB) was highest in the case of the trimer (-37.4 Kcal/mol), followed by the dimer (-21.2 Kcal/mol) and monomer (-18.2 Kcal/mol). Dimer obtained the highest glide score of 8.25 (FEB =-25.1 Kcal/mol) with p67phox-isoform 2. The PH domain of p47phox and the SH3 domain of p67phox have their own best binding energy with dimmer. While molecular docking with Rac-Zn-GD, P, dimer, and trimer have shown comparable FEB. The residues, on which the ligands were found to interact, were of major significance being present in those domains that vicinity to inhibit or activate the complex and are important for the protein structure and functioning. MDS studies have confirmed the findings that the Apocynin trimer molecule has superior stability and interactions with the enzyme complex. Conclusion: It can be concluded from the study that trimer and dimer have better interactions in terms of FEB with p67phox and p47phox, indicating the reported findings in the literature.
背景/目的:一种名为NADPH氧化酶(NOX)的多组分酶系统通过产生活性氧(ROS)来帮助先天免疫。NOX过度活化与多种疾病有关。这种酶是一种膜结合的复合物,在活性时由六个亚基组成。这些酶亚基被认为是酶活性的有效抑制剂和减少氧化应激的良好靶点。方法:本研究旨在使用计算机工具分析可能的靶标:NOX的不同亚基,以及它们与罗布麻素的相互作用,以确定其对NOX的可能抑制机制。罗布麻素的单体、二聚体和三聚体与NOX的不同亚基对接。结果:当罗布麻素的单体和二聚体与NOX的p47phox完全亚基对接时,获得了可比较的滑动得分,并且比三聚体更好。在三聚体(-37.4Kcal/mol)的情况下,结合自由能(FEB)最高,其次是二聚体(-21.2Kcal/mmol)和单体(-18.2Kcal/mmol)。二聚体与p67phox同种型2的滑翔得分最高,为8.25(FEB=-25.1Kcal/mol)。p47phox的PH结构域和p67phox的SH3结构域具有各自的最佳结合能和二聚体。虽然与Rac-Zn-GD、P、二聚体和三聚体的分子对接显示出相当的FEB。发现配体在其上相互作用的残基存在于那些邻近的结构域中,具有抑制或激活复合物的重要意义,并且对蛋白质结构和功能很重要。MDS研究证实了夹竹桃素三聚体分子具有优异的稳定性以及与酶复合物的相互作用。结论:从研究中可以得出结论,三聚体和二聚体在FEB方面与p67phox和p47phox具有更好的相互作用,表明了文献中报道的结果。
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引用次数: 0
Role of Nutritional Supplements on Gut-Muscle Axis Across Age: a Mini-Review. 营养补充剂对肠道-肌肉轴的作用:一项小型综述。
Q3 CELL BIOLOGY Pub Date : 2023-05-14 DOI: 10.33594/000000628
Ricardo Aparecido Baptista Nucci, Victor Abou Nehmi Filho, Wilson Jacob-Filho, José Pinhata Otoch, Ana Flávia Marçal Pessoa

Sarcopenia is a progressive skeletal muscle disorder associated with aging, resulting in loss of muscle mass and function. It has been linked to inflammation, oxidative stress, insulin resistance, hormonal changes (i.e. alterations in the levels or activity of hormones which can occur due to a variety of factors, including aging, stress, disease, medication, and environmental factors), and impaired muscle satellite cell activation. The gut microbiome is also essential for muscle health, and supplements such as probiotics, prebiotics, protein, creatine, and betaalanine can support muscle growth and function while also promoting gut health. Chronic low-grade inflammation is a leading cause of sarcopenia, which can activate signaling pathways that lead to muscle wasting and reduce muscle protein synthesis. Insulin resistance, hormonal changes, and impaired muscle satellite cell activation contribute to sarcopenia, and high levels of fat mass also play a role in the pathogenesis of sarcopenia. Resistance exercise and dietary supplementation have been shown to be effective treatments for sarcopenia. In addition, a combination of resistance exercise and supplementation has been shown to have a more significant beneficial effect on anthropometric and muscle function parameters, leading to a decrease in sarcopenic state. Thus, understanding the relationship between the gut microbiome and muscle metabolism is crucial for developing new treatments for sarcopenia across age groups.

肌少症是一种与衰老相关的进行性骨骼肌疾病,导致肌肉质量和功能的丧失。它与炎症、氧化应激、胰岛素抵抗、激素变化(即由于各种因素,包括衰老、压力、疾病、药物和环境因素,可能发生的激素水平或活性的改变)以及肌肉卫星细胞激活受损有关。肠道微生物群对肌肉健康也是必不可少的,益生菌、益生元、蛋白质、肌酸和β -丙氨酸等补充剂可以支持肌肉生长和功能,同时也促进肠道健康。慢性低度炎症是肌肉减少症的主要原因,它可以激活导致肌肉萎缩和减少肌肉蛋白质合成的信号通路。胰岛素抵抗、激素变化和肌肉卫星细胞激活受损可导致肌肉减少症,高水平的脂肪量也在肌肉减少症的发病机制中发挥作用。抗阻运动和膳食补充已被证明是有效的治疗肌肉减少症。此外,抗阻运动和补充相结合已被证明对人体测量和肌肉功能参数有更显著的有益影响,导致肌肉减少状态的减少。因此,了解肠道微生物组和肌肉代谢之间的关系对于开发跨年龄组肌肉减少症的新疗法至关重要。
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引用次数: 3
Expression of Concern. 表达关心。
Q3 CELL BIOLOGY Pub Date : 2023-04-30 DOI: 10.33594/000000624
{"title":"Expression of Concern.","authors":"","doi":"10.33594/000000624","DOIUrl":"https://doi.org/10.33594/000000624","url":null,"abstract":"","PeriodicalId":9845,"journal":{"name":"Cellular Physiology and Biochemistry","volume":"57 2","pages":"158"},"PeriodicalIF":0.0,"publicationDate":"2023-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9396645","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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Cellular Physiology and Biochemistry
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