Inhibitory actions of mibefradil on steroidogenesis in mouse Leydig cells： involvement of Ca^2＋ entry via the T-type Ca^2＋ channel

Intracellular cAMP and Ca^2＋ are involved in the regulation of steroidogenic activity in Leydig cells, which coordinate responses to luteinizing hormone （LH） and human ehorionic gonadotropin （hCG）. However, the identification of Ca^2＋ entry implicated in Leydig cell steroidogenesis is not well defined. The objective of this study was to identify the type of Ca^2＋ channel that affects Leydig cell steroidogenesis. In vitro steroidogenesis in the freshly dissociated Leydig cells of mice was induced by hCG incubation. The effects of mibefradil （a putative T-type Ca^2＋ channel blocker） on steroidogenesis were assessed using reverse transcription （RT）-polymerase chain reaction analysis for the steroidogenic acute regulatory protein （STAR） mRNA expression and testosterone production using radioimmunoassay. In the presence of 1.0 mmol L-1 extracellular Ca^2＋, hCG at 1 to 100 IU noticeably elevated both StAR mRNA level and testosterone secretion （P 〈 0.05）, and the stimulatory effects of hCG were markedly diminished by mibefradil in a dose-dependent manner （P 〈 0.05）. Moreover; the hCG-induced increase in testosterone production was completely removed when external Ca^2＋ was omitted, implying that Ca entry is needed for hCG-induced steroidogenesis. Furthermore, a patch-clamp study revealed the presence of mibefradil-sensitive Ca^24- currents seen at a concentration range that nearly paralleled those inhibiting steroidogenesis. Collectively, Our data provide evidence that hCG-stimulated steroidogenesis is mediated at least in part by Ca^2＋ entry carried out by the T-type Ca^2＋ channel in the Leydig cells of mice.

富里酸和Ca^(2+)共存对嗜酸性氧化亚铁硫杆菌介导生成次生高铁矿物的影响

为揭示富里酸和Ca^(2+)共存对嗜酸性氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans)氧化酸性矿山废水(AMD)中的Fe^(2+)和形成次生高铁矿物的影响,分析了pH、Fe^(2+)氧化率、铁沉淀率以及次生高铁矿物矿相、基团等相关指标。结果表明,Ca^(2+)确实具有提高嗜酸性氧化亚铁硫杆菌氧化Fe^(2+)的能力。低质量浓度(0.2 g/L)的富里酸对嗜酸性氧化亚铁硫杆菌活性的提高具有促进作用,高质量浓度(0.4 g/L)的富里酸具有抑制作用,而增加Ca^(2+)反过来能够减弱高浓度富里酸对嗜酸性氧化亚铁硫杆菌的抑制作用。对形成的次生高铁矿物进行X射线衍射(XRD)和傅立叶红外光谱(FTIR)分析,结果表明高浓度富里酸促进了另一次生高铁矿物草黄铁矾的生成。

川芎嗪抑制ROCK的表达降低模拟失重大鼠血管Ca^(2+)敏感性

研究失重条件下血管平滑肌收缩性、Ca^(2+)敏感性及其调控通路RhoA-ROCK蛋白表达的变化,以及川芎嗪干预对其的影响。大鼠尾部悬吊模拟失重,在机体前、后部分别选取颈总动脉和肠系膜上动脉。在模拟失重大鼠颈总动脉中,由苯肾上腺素(PHE)或KCl诱发的血管收缩性和Ca^(2+)敏感性增强,RhoA激酶2(ROCK II)的表达、肌球蛋白磷酸酶靶亚基1(MYPT1)和肌球蛋白调节轻链(MLC)的磷酸化水平均上升,血管孵育Y-27632(ROCK特异性抑制剂)后可降低以上变化。模拟失重大鼠灌饲川芎嗪亦可降低以上变化。模拟失重后,大鼠肠系膜上动脉的收缩性和Ca^(2+)敏感性、ROCK II的表达、MYPT1和MLC的磷酸化水平降低,血管孵育Y-27632对以上变化无明显作用。模拟失重大鼠灌饲川芎嗪亦对以上变化无明显作用。结果表明,由RhoA-ROCK调控的血管平滑肌Ca^(2+)敏感性的变化可能是失重条件下机体前后部血管收缩性发生区域性重塑的关键因素。川芎嗪可抑制ROCK蛋白的表达,降低血管平滑肌升高的Ca^(2+)敏感性,从而纠正失重条件下机体前部血管收缩性的增强,但对失重条件下机体后部血管收缩性的减弱无恢复作用。

外源Ca^(2+)对玉米幼苗镉胁迫的缓解效应

本试验以玉米为研究材料,对玉米幼苗进行一定浓度的镉(20mg/L)胁迫,通过对玉米幼苗施加不同浓度的Ca^(2+)(0、2、5、8mmol/L),研究Ca^(2+)对镉胁迫下玉米幼苗的叶绿素、MDA、可溶性糖、可溶性蛋白、游离脯氨酸和POD活性6个生理生化指标的影响。结果表明:施加外源Ca^(2+)显著增加了镉胁迫下南瓜幼苗叶片中叶绿素含量(P<0.05),而MDA含量、可溶性糖含量、游离脯氨酸含量以及POD活性均显著降低(P<0.05),对可溶性蛋白的缓解作用不显著(P>0.05)。其中以5mmol/LCa^(2+)的缓解效果最好。

复合混凝剂中Ca^(2+)对高溶解态磷坑塘水混凝效果的影响

文章以Ca^(2+)改性后的AlCl_(3)和PACl作为混凝剂进行混凝实验,通过出水浊度、余铝、总磷、絮体粒径及有机物组成等分析改性后混凝剂的作用机理,探讨混凝剂中存在的Ca^(2+)对高磷坑塘水混凝效果的影响机制。浊度去除方面,低投加量下Ca^(2+)可明显降低出水浊度。当投加量为0.10 mmol/L时,PACl出水浊度下降了33.52 NTU,AlCl_(3)出水浊度下降了28.72 NTU。余铝去除方面,当AlCl_(3)作混凝剂时,Ca^(2+)可以通过增加混凝剂的电中和能力来降低出水余铝浓度。溶解态磷去除方面,Ca^(2+)与磷酸根反应或通过压缩双电层和吸附电中和作用来增加溶解态磷的去除率。当投加量为0.20 mmol/L时,Ca^(2+)浓度为0.9 mmol/L,PACl溶解态磷去除率较未改性前提高16.1%。絮体粒径方面,Ca^(2+)可以促进颗粒之间脱稳凝聚,增加絮体粒径和分形维数,AlCl_(3)在0.20 mmol/L投加量下絮体粒径增加79μm。并且Ca^(2+)的加入使AlCl_(3)生成的絮体抗剪切能力更强,但是絮体受到破坏后更不容易恢复,使PACl形成絮体强度因子和恢复因子变大。有机物去除方面,Ca^(2+)可以提高有机物的去除率。对于0.20 mmol/L投加量,Ca^(2+)为0.06 mmol/L投加量条件下AlCl_(3)改性后混凝剂荧光响应值由1100降至800。

Ca^(2+)/CaN/NFAT信号通路在肿瘤中的研究进展

肿瘤成为危害人类健康的主要原因,其发生发展与免疫炎症密切相关.Ca^(2+)/CaN/NFAT信号通路是目前已知的免疫炎症通路,可促进多种免疫细胞活化,介导免疫炎症因子释放,进而影响机体的免疫功能.目前,Ca^(2+)/CaN/NFAT信号通路在肿瘤中的存在和作用越来越受关注.多项研究表明与该通路与肿瘤发生发展密切相关.故本文从Ca^(2+)/CaN/NFAT信号通路概述、信号通路对肿瘤发生发展的影响、信号通路对肿瘤免疫炎症的作用及与信号通路相关药物在防治肿瘤中的作用四方面进行综述,以期为临床防治肿瘤提供新视角新靶点.

Ca^(2+)-ATPase参与植物耐盐性调控的研究进展

盐胁迫下细胞质Ca^(2+)浓度升高,细胞会激活Ca^(2+)调节的靶酶或者与Ca^(2+)高度亲和的受体蛋白,其中,与Ca^(2+)高度亲和的受体蛋白中,植物钙泵(Ca^(2+)-ATPase)是P型ATP酶,包含内质网Ca^(2+)-ATPases与质膜Ca^(2+)-ATPas‐es,通过主动运输将Ca^(2+)从细胞质转移到质外体或细胞器。大量研究表明,植物的耐盐性在很大程度上与其维持钙泵即Ca^(2+)-ATPase活性的能力有关。多种植物Ca^(2+)-ATPase对盐胁迫表现出敏感性,并受到外源Ca^(2+)的保护,表明外源钙处理与Ca^(2+)-ATPase活性可能在盐胁迫下的细胞内钙稳态和信号转导中起重要作用。该研究概述了植物Ca^(2+)-ATPase类型、结构与性质,亚细胞定位Ca^(2+)-ATPase及外源钙与亚细胞定位Ca^(2+)-ATPase参与植物耐盐调控研究进展,重点对质膜、液泡膜、核膜、内质网及高尔基体Ca^(2+)-ATPases参与植物耐盐调控的研究进展进行了综述,并提出展望。该研究为了解植物耐盐性生理及分子机制提供帮助,同时为作物耐盐栽培提供新思路。

β-细辛醚通过抑制TRPV4的表达缓解谷氨酸诱导的Ca^(2+)超载

谷氨酸处理会导致Ca^(2+)超载,瞬时受体电位香草素受体4(transient receptor potential vanilloid 4,TRPV4)在其中的作用及可能机制尚不清楚。β-细辛醚能快速透过血脑屏障,对兴奋性毒性具有较强的神经保护作用。文章以高分化的PC12细胞为研究对象,探究β-细辛醚(15、30、60μmol/L)预处理4 h,40 mmol/L谷氨酸处理实时记录对PC12细胞Ca^(2+)浓度的影响,采用钙成像技术检测Ca^(2+)浓度的变化;采用实时荧光定量聚合酶链式反应(polymerase chain reaction,PCR)、Western Blot及免疫荧光技术检测TRPV4的mRNA和蛋白的表达;采用Lipofectiamine 2000脂质体实验转染TRPV4-siRNA和pEX-3-TRPV4,观察沉默和过表达TRPV4对谷氨酸引起Ca^(2+)超载的影响。结果表明:与正常对照组相比,谷氨酸处理5 min可诱导Ca^(2+)超载,显著提高TRPV4的mRNA和蛋白的表达;与模型组相比,β-细辛醚能够剂量依赖性地降低谷氨酸诱导的Ca^(2+)超载和TRPV4的表达;沉默TRPV4抑制细胞Ca^(2+)超载;过表达TRPV4则部分逆转β-细辛醚抑制谷氨酸诱导的Ca^(2+)超载。该研究证明,谷氨酸处理PC12细胞5 min通过上调TRPV4的表达诱导Ca^(2+)超载,β-细辛醚作为TRPV4的拮抗剂,是一种潜在的抑制兴奋性毒性的药物。

活血益肾法联合西药对踝关节骨折术后血清Ca^(2+)、ALP及踝关节功能的影响

目的探讨活血益肾法联合西药对踝关节骨折术后血清Ca^(2+)、碱性磷酸酶(ALP)及踝关节功能的影响。方法选取踝关节骨折术后患者105例,随机分为试验组(n=53)和对照组(n=52)。对照组给予地塞米松治疗,试验组在对照组的基础上加用活血益肾法治疗。比较两组临床疗效、血清Ca^(2+)、ALP、踝关节功能水平变化情况、临床症状改善情况及不良反应发生情况。结果治疗后,试验组踝关节功能恢复优良率显著高于对照组;治疗后,试验组血清Ca^(2+)、ALP水平高于对照组;治疗后,两组肌力背伸、跖屈和背伸关节主动活动度、跖屈关节主动活动度水平均升高,且试验组高于对照组;治疗后,试验组临床各症状改善时间均显著低于对照组;以上组间差异均有统计学意义(P<0.05)。治疗期间,两组不良反应比较差异无统计学意义(P>0.05)。结论踝关节骨折术后应用活血益肾法联合西药效果显著,可有效改善患者血清Ca^(2+)、ALP及踝关节功能水平。

miR-361介导PI3K/Akt/Ca^(2+)通路对大鼠肥大细胞增殖情况及组胺与IL-4表达水平的影响

目的:探讨miR-361介导PI3K/AKt/Ca^(2+)通路对活化大鼠肥大细胞增殖情况及组胺与白细胞介素-4(IL-4)表达水平的影响。方法:将大鼠肥大细胞(RBL-2H3细胞)分为miR-361过表达组、miR-361抑制组、miR-361对照组和正常对照组,miR-361过表达组、miR-361抑制组、miR-361对照组细胞分别转染miR-361过表达、miR-361抑制及miR-361无义序列寡核苷酸片段,测定并比较各组细胞miR-361 mRNA、组胺、IL-4及PI3K/AKt/Ca^(2+)通路相关蛋白水平。结果:miR-361过表达组细胞miR-361 mRNA相对表达水平高于miR-361抑制组、miR-361对照组及正常对照组,差异均有统计学意义(P<0.05)。CCK-8法检测结果显示,miR-361过表达组24、48、72 h细胞增值率均低于miR-361抑制组、miR-361对照组及正常对照组,差异均有统计学意义(P<0.05)。ELISA法检测结果显示,miR-361过表达组细胞组胺和IL-4水平的表达水平低于miR-361抑制组、miR-361对照组及正常对照组,差异均有统计学意义(P<0.05)。Western印迹法检测结果显示,miR-361过表达组细胞p-AKT、AKT、p-PI3K、PI3K蛋白表达水平均高于miR-361抑制组、miR-361对照组及正常对照组,差异均有统计学意义(P<0.05)。结论:miR-361过表达可抑制大鼠肥大细胞的增殖,降低组胺与IL-4的表达水平,其作用机制可能与其能够提高PI3K/AKt/Ca^(2+)通路活性有关。

STIM1和Orai1/TRPCs在右心衰竭Ca^(2+)重塑中的作用

本文总结右心衰竭心肌细胞动作电位变化、心肌细胞内T管和内质网的改变,详细阐述了钙调神经磷酸酶(Calcineurin)-活化细胞活因子(NFAT)通路对心肌收缩力的作用。重点描述基质相互作用因子1(STIM1)和钙释放激活钙通道蛋白1(Orai1)/瞬时受体电位通道(TRPCs)通路在右心室心肌细胞兴奋收缩耦联、肥大及右心室成纤维细胞迁移增强方面的机制。

Ca^(2+)对茅苍术丛生芽诱导及生理特征的影响

为提高茅苍术组培苗品质,以信阳野生茅苍术种子获取的初代无菌试管苗为试材,采用植物组织培养试验方法,研究不同浓度Ca^(2+)对茅苍术丛生芽诱导及生理特征的影响。结果表明:Ca^(2+)浓度为1.50 mmol/L时茅苍术丛生芽增殖数最高,为16.36个;Ca^(2+)浓度为3.00 mmol/L时丛生芽总干重最高,为0.37 g;Ca^(2+)浓度为6.00 mmol/L时丛生芽总鲜重、单芽平均叶片数最高,分别为3.19 g、5.44个;Ca^(2+)浓度为12.00 mmol/L时茅苍术丛生芽的株高最高,为493.95 mm;Ca^(2+)浓度为24.00 mmol/L时叶绿素a含量、叶绿素b含量、类胡萝卜素含量、叶绿素总量和丛生芽SOD总活性最高,分别为1.54 mg/g、0.40 mg/g、0.36 mg/g、1.94 mg/g和329.36 U/g。在茅苍术组织培养过程中可根据不同需求选择相应的Ca^(2+)浓度。

Synergistic strengthening mechanism of Ca^(2+)-sodium silicate to selective separation of feldspar and quartz

Inhibitors are important for flotation separation of quartz and feldspar.In this study,a novel combined inhibitor was used to separate quartz and feldspar in near-neutral pulp.Selective inhibition of the combined inhibitor was assessed by micro-flotation experiments.And a series of detection methods were used to detect differences in the surface properties of feldspars and quartz after flotation reagents and put forward the synergistic strengthening mechanism.The outcomes were pointed out that pre-mixing combined inhibitors were more effective than the addition of Ca^(2+)and SS in sequence under the optimal proportion of 1:5.A concentrate from artificial mixed minerals that was characterized by a high quartz grade and a high recovery was acquired,and was found to be 90.70wt% and 83.70%,respectively.It was demonstrated that the combined inhibitor selectively prevented the action of the collector and feldspar from Fourier-transform infrared(FT-IR)and adsorption capacity tests.The results of X-ray photoelectron spectroscopy(XPS)indicated that Ca^(2+)directly interacts with the surface of quartz to increase the adsorption of collectors.In contrast,the chemistry property of Al on the feldspar surface was altered by combined inhibitor due to Na^(+)and Ca^(2+)taking the place of K^(+),resulting in the composite inhibitor forms a hydrophilic structure,which prevents the adsorption of the collector on the surface of feldspar by interacting with the Al active site.The combination of Ca^(2+)and SS synergically strengthens the difference of collecting property between quartz and feldspar by collector,thus achieving the effect of efficient separation.A new strategy for flotation to separate quartz from feldspar in near-neutral pulp was provided.

Performance and enhanced oil recovery efficiency of an acid-resistant polymer microspheres of anti-CO_(2) channeling in low-permeability reservoirs

CO_(2) flooding is a vital development method for enhanced oil recovery in low-permeability reservoirs,However,micro-fractures are developed in low-permeability reservoirs,which are essential oil flow channels but can also cause severe CO_(2) gas channeling problems.Therefore,anti-gas channeling is a necessary measure to improve the effect of CO_(2) flooding.The kind of anti-gas channeling refers to the plugging of fractures in the deep formation to prevent CO_(2) gas channeling,which is different from the wellbore leakage.Polymer microspheres have the characteristics of controllable deep plugging,which can achieve the profile control of low-permeability fractured reservoirs.In acidic environments with supercritical CO_(2),traditional polymer microspheres have poor expandability and plugging properties.Based on previous work,a systematic evaluation of the expansion performance,dispersion rheological properties,stability,deep migration,anti-CO_(2) channeling and enhanced oil recovery ability of a novel acid-resistant polymer microsphere(DCNPM-A)was carried out under CQ oilifield conditions(salinity of85,000 mg/L,80℃,pH=3).The results show that the DCNPM-A microsphere had a better expansion performance than the traditional microsphere,with a swelling rate of 13.5.The microsphere dispersion with a concentration of 0.1%-0.5%had the advantages of low viscosity,high dispersion and good injectability in the low permeability fractured core.In the acidic environment of supercritical CO_(2),DCNPM-A microspheres showed excellent stability and could maintain strength for over 60 d with less loss.In core experiments,DCNPM-A microspheres exhibited delayed swelling characteristics and could effectively plug deep formations.With a plugging rate of 95%,the subsequent enhanced oil recovery of CO_(2) flooding could reach 21.03%.The experimental results can provide a theoretical basis for anti-CO_(2)channeling and enhanced oil recovery in low-permeability fractured reservoirs.

A Research Progress of CO_(2)-Responsive Plugging Channeling Gels

In the heterogeneous reservoirs,CO_(2) flooding easily leads to CO_(2) gas channeling,which can seriously affect sweeping efficiency and reduce oil recovery.After thoroughly investigating the advantages and shortcomings of various CO_(2) plugging technologies,this paper focuses on the feasibility of improving conventional water-alternating gas(WAG)through CO_(2)-responsive gel materials.Based on the different chemical reaction mechanisms between the unique chemical structure and CO_(2),changes in the material’s physical and chemical properties can respond to CO_(2).The feasibility of utilizing these property changes for CO_(2)-responsive plugging is explored.Various CO_(2)-responsive gels and gel nanoparticles have been extensively researched in different fields,such as energy,medicine,and biology.This paper surveys the molecular structures,chemical compositions,response mechanisms,and changes of these CO_(2)-responsive gels,aiming to draw insights into the carbon dioxide-enhanced oil recovery(CO_(2)-EOR)field.Finally,the key issues and future development direction of CO_(2)-responsive plugging gels were analyzed.

外源Ca^(2+)对两种产脲酶细菌修复Cd-As复合污染水稻土的影响

微生物诱导碳酸盐沉淀(Microbially induced carbonate precipitation,MICP)技术已被广泛应用于土壤重金属污染修复。为促进MICP过程,提高土壤修复效果,以Cd-As复合污染的水稻土为研究对象,利用巴氏八叠球菌(Octococcus pasteurii)和蜡样芽孢杆菌(Bacillus cereus)两种产脲酶细菌,分析比较了外源添加氯化钙(CaCl2)对两种菌株固定土壤中Cd、As效果的影响,并对修复后土壤理化性质、酶活性及微生物多样性的变化进行了检测。结果表明:两种菌株均能固定土壤Cd、As,其中,蜡样芽孢杆菌对土壤Cd、As的固定效果更佳,与巴氏八叠球菌处理相比,蜡样芽孢杆菌处理下土壤有效态Cd、As含量分别降低了16.7%、11.1%;添加外源Ca^(2+)后,在两种细菌处理下有效态Cd、As含量均发生显著变化,分别降低了17.3%~22.2%、16.8%~26.7%,可见Ca^(2+)的添加能有效促进MICP过程,促进对Cd、As的固定。此外,与未添加Ca^(2+)处理相比,添加Ca^(2+)后,两种细菌处理下显著提高了土壤脲酶活性(52.6%~113.3%)、蔗糖酶活性(13.1%~28.9%)、碱解氮含量(3.4%~25.5%)、速效钾含量(2.1%~34.1%)以及微生物多样性,表明外源Ca^(2+)可有效提高土壤肥力及土壤生态功能。综上,基于MICP作用,可通过添加外源Ca^(2+)来增强产脲酶细菌对Cd-As复合污染土壤的修复效果,其中,蜡样芽孢杆菌修复效果更佳。

氧糖剥夺再灌注对神经元Wnt5a/FZD2/Ca^(2+)通路的影响

目的:探讨氧糖剥夺再灌注(OGD/Rep)后神经元细胞内Wnt5a/FZD2/Ca^(2+)通路的变化。方法:采用OGD/Rep模型构建神经元细胞局部缺血/再灌注损伤,分正常组、再灌注(0、3、6、9、12、24 h)组。免疫荧光鉴定皮层神经元细胞,CCK-8法确定神经元细胞OGD时间,Western blotting检测非经典Wnt/Ca^(2+)信号通路中Wnt5a、FZD2、IP3-R和p-CaMKⅡ/CaMKⅡ蛋白的变化,免疫荧光和酶标仪检测细胞内Ca^(2+)和活性氧(ROS)的含量。结果:MAP-2免疫荧光鉴定原代皮层神经元细胞纯度较高。与正常组比较,神经元细胞OGD 3 h后细胞存活率为55.46%;与正常组比较,OGD/Rep后非经典Wnt/Ca^(2+)信号通路相关蛋白Wnt5a(3、6、9、12、24 h)、FZD2(3、6、9 h)、IP3-R(6、9、12、24 h)和p-CaMKⅡ/CaMKⅡ(6 h)的相对表达量上调(P<0.05~P<0.01);与正常组比较,OGD/Rep后神经细胞内Ca^(2+)和ROS的表达量均升高(P<0.01)。结论:神经元细胞OGD/Rep能激活Wnt5a/FZD2/Ca^(2+)信号通路。

Tensor Decomposition-Based Channel Estimation and Sensing for Millimeter Wave MIMO-OFDM V2I Systems

An integrated sensing and communication(ISAC)scheme for a millimeter wave(mmWave)multiple-input multiple-output orthogonal frequency division multiplexing(MIMO-OFDM)Vehicle-to-Infrastructure(V2I)system is presented,in which both the access point(AP)and the vehicle are equipped with large antenna arrays and employ hybrid analog and digital beamforming structures to compensate the path loss,meanwhile compromise between hardware complexity and system performance.Based on the sparse scattering nature of the mmWave channel,the received signal at the AP is organized to a four-order tensor by the introduced novel frame structure.A CANDECOMP/PARAFAC(CP)decomposition-based method is proposed for time-varying channel parameter extraction,including angles of departure/arrival(AoDs/AoAs),Doppler shift,time delay and path gain.Then leveraging the estimates of channel parameters,a nonlinear weighted least-square problem is proposed to recover the location accurately,heading and velocity of vehicles.Simulation results show that the proposed methods are effective and efficient in time-varying channel estimation and vehicle sensing in mmWave MIMOOFDM V2I systems.

干姜吴茱萸水煎液对大鼠结肠平滑肌细胞L型Ca^(2+)通道的影响

目的:探讨干姜吴茱萸水煎液对大鼠结肠平滑肌收缩及平滑肌细胞L型Ca^(2+)通道的影响机制。方法:采用0、0.01、0.1、1、10 g/L干姜吴茱萸(1∶1)水煎液依次作用于正常结肠平滑肌,10 g/L水煎液及L型Ca^(2+)通道激动剂BAY-K-8644依次作用于氯化乙酰胆碱(Ach)诱导后的正常平滑肌,检测平滑肌收缩张力、频率和振幅;全细胞膜片钳记录L型钙电流;Western blot检测各组细胞中L-型电压依赖钙离子通道α1C亚基(α1c)和钙调蛋白(CaM)蛋白表达情况。结果:1、10 g/L水煎液组能显著降低离体结肠平滑肌的收缩张力(P<0.05),对频率及振幅并无显著影响(P>0.05);与0 g/L水煎液组相比,Ach促进平滑肌收缩张力、频率和振幅显著增加(P<0.05);与Ach组相比,10 g/L水煎液显著降低平滑肌的收缩张力、频率和振幅(P<0.05);与10 g/L水煎液组相比,BAY-K-8644显著增加平滑肌的收缩张力、频率和振幅(P<0.05);与0 g/L水煎液组相比,Ach可显著增加ICa-L峰值电流密度(P<0.05);与Ach组相比,10 g/L水煎液显著降低ICa-L峰值电流密度(P<0.05);与10 g/L水煎液组相比,BAY-K-8644可显著增加ICa-L峰值电流密度(P<0.05);与对照组相比,Ach组结肠平滑肌细胞Ca^(2+)相对荧光强度峰值、α1c、CaM表达显著增加(P<0.05);Ach组相比,10 g/L组Ca^(2+)相对荧光强度峰值、α1c和CaM表达水平显著降低(P<0.05);与10 g/L组相比,BAYK-8644组Ca^(2+)相对荧光强度峰值、α1c、CaM表达显著增加(P<0.05)。结论:干姜吴茱萸水煎液可抑制大鼠结肠平滑肌收缩,并阻滞L型Ca^(2+)通道。

基于Ca^(2+)-CaMKⅡ信号通路的济川煎治疗阳虚便秘的作用及分子机制

目的基于Ca^(2+)-CaMKⅡ通路观察济川煎对阳虚便秘大鼠的治疗作用及分子机制。方法采用白醋+活性炭冰水联合复方地芬诺酯片复制阳虚便秘大鼠模型。将模型成功大鼠随机分为5组,即模型组、莫沙必利组(1.88 mg·kg^(-1))、济川煎高(8.26 g·kg^(-1))、中(4.31 g·kg^(-1))、低(2.16 g·kg^(-1))剂量组,灌胃给药每日1次,连续7 d。末次给药,记录大鼠状态并评分,进行排便功能测定;分离模型大鼠结肠平滑肌细胞并鉴定,FCM检测结肠平滑肌细胞内Ca^(2+)浓度变化;采用ELISA法检测大鼠血清中cAMP、cGMP含量及其比值;采用RT-PCR及Western blot法检测大鼠结肠平滑肌细胞CaM、CaMKⅡ亚型的表达情况。结果济川煎可明显缓解模型大鼠腹胀,摄食、饮水量、排便减少及体质量降低的症状(P<0.05),评分明显升高(P<0.01);能明显缩短首粒排便时间(P<0.01),增加排便粒数(P<0.05);济川煎还可明显增加模型大鼠血清中cAMP含量(P<0.05),减少cGMP含量(P<0.01),cAMP/cGMP比值明显增加(P<0.01);明显降低大鼠结肠平滑肌细胞内Ca^(2+)浓度(P<0.01);PCR及Western blot检测结果显示,济川煎可明显降低CaM及CaMKⅡβ、γ、δ亚型蛋白表达(P<0.01)。结论济川煎对阳虚便秘具有明显的治疗作用,可能与其调节Ca^(2+)-CaMKⅡ信号通路有关。