二甲双胍对华法令诱导大鼠动脉钙化的影响及其机制初探

目的:研究二甲双胍(metformin,MET)对华法令(Warfarin,WFN)诱导大鼠动脉钙化的影响及其机制。方法:将28只SD大鼠随机分为正常对照组、8周(W)钙化组、8W钙化+8W MET 100 mg/kg治疗组、8W钙化+8W MET 200 mg/kg治疗组。采用Von Kossa染色法检测胸主动脉组织中钙结节;邻甲酚肽络合酮比色法测定颈总动脉组织中钙沉积含量;免疫组化染色检测血管壁中成骨基因Runx2及血管平滑肌标志物α-SMA的表达;Western Blot检测血管壁中Runx2及自噬标志物LC3II的表达。结果:WFN干预8 W后,大鼠动脉中钙沉积含量显著增加(P0.01),MET治疗组主动脉钙含量与钙化组相比均显著降低(P0.01)。Von Kossa染色可见钙化组(WFN组)动脉壁中层黑色连续钙盐沉积条带,而进行MET治疗后,黑色条带明显减少,对照组未见黑色条带。免疫组化显示钙化组血管壁Runx2表达阳性,棕褐色染色较深,而α-SMA表达则显著下降,基本未见棕褐色染色沉积;MET治疗后能够逆转上述趋势。Western Blot显示钙化组血管壁Runx2表达明显上升,MET治疗后Runx2表达被抑制。此外,钙化过程中伴随着自噬标志物LC3II表达轻度上升;随着MET浓度升高,血管壁中自噬水平呈剂量依赖性显著升高。结论:二甲双胍能够有效抑制大鼠动脉钙化,减轻血管平滑肌细胞由收缩表型向成骨样表型转换,其机制可能与诱导自噬有关。     

慢性肾功能衰竭继发高尿酸血症促进血管钙化

本文旨在探讨高尿酸血症对肾衰血管钙化的影响及其机制。使用腺嘌呤饮食喂养的方法建立慢性肾功能衰竭的大鼠模型。将8周龄雄性Wistar大鼠随机分为对照组(正常饮食+正常饮水)、肾衰组(腺嘌呤饮食+正常饮水)和肾衰+别嘌呤醇组(腺嘌呤饮食+别嘌呤醇饮水),喂养6周后进行取材,用邻甲酚酞络合铜法和Von Kossa染色法检测大鼠腹主动脉钙含量,用real-time PCR法检测大鼠腹主动脉成骨基因(Cbfα1、Msx2、Osx和Sox9)、平滑肌细胞标志物(SM22a和Acta2)和钙化抑制因子Opn和Mgp的m RNA表达水平。结果显示,肾衰组大鼠血磷、血肌酐、血清尿素氮和血尿酸水平均显著升高,别嘌呤醇治疗可显著降低肾衰大鼠血肌酐、血清尿素氮和血尿酸水平,但是不影响血磷水平。肾衰组大鼠的腹主动脉钙含量显著高于对照组(P0.05),肾衰+别嘌呤醇组显著低于肾衰组(P0.05);肾衰组大鼠腹主动脉Cbfα1、Msx2、Osx和Sox9的m RNA水平较对照组出现显著上调,而SM22a、Acta2、Mgp和Opn的m RNA水平出现下调,在肾衰+别嘌呤醇组Msx2、Osx、SM22a和Opn的m RNA表达变化被逆转。在高磷(3 mmol/L)诱导的血管平滑肌细胞(vascular smooth muscle cells,VSMC)钙化模型中,抗氧化剂Mn TMPy P显著抑制VSMC钙化,而别嘌呤醇则无显著作用;当尿酸水平达到6和7 mg/d L时可以显著加重高磷诱导的VSMC钙化(P0.05)。以上结果提示,慢性肾功能衰竭继发的高尿酸血症可以促进VSMC向成骨/成软骨样细胞转分化,进而加剧血管钙化的发生。     

普罗布考对动脉粥样硬化大鼠平滑肌细胞凋亡及p53和Fas蛋白表达的影响

目的探讨普罗布考防治动脉粥样硬化（AS）的机制。方法选用雄性大鼠,复制大鼠AS模型,随机分为动脉粥样硬化模型组、普罗布考组和正常对照组。大鼠造模成功后给予普罗布考治疗,6周后处死大鼠,采用流式细胞术检测平滑肌细胞凋亡率及凋亡相关基因p53和Fas蛋白的表达。结果模型组大鼠血管平滑肌细胞凋亡率明显高于对照组（P〈0．05）,p53和Fas蛋白的表达增强（P％0．05）,主动脉壁可肉眼观测典型斑块。普罗布考组大鼠平滑肌细胞的凋亡率明显低于模型组（P〈0．05）,p53和Fas蛋白表达下调（P〈0．05）,主动脉斑块面积较模型组减小明显。结论普罗布考通过调节p53和Fas蛋白表达来调节AS大鼠平滑肌细胞的凋亡。     

运动训练对大鼠主动脉应力和NF-κB及c-fos表达的影响

目的：探讨不同强度运动训练对大鼠主动脉应力和NF-κB及c-fos表达的影响。方法：采用跑台训练方式,建立大鼠有氧运动和疲劳运动模型,运用免疫组织化学SABC法研究不同强度运动训练对大鼠主动脉VEC和VSMC中NF-κB及c-fos表达的影响。结果：胸主动脉血压的变化与对照组比较,有氧训练和疲劳训练均显著性升高（P〈0．05）,但有氧训练与疲劳训练之间差异不显著。与对照组比较,有氧训练大鼠VEC和VSMC中NF-κB和c-fos均显著性下调表达,胸主动脉张开角显著性升高（P〈0．05）,疲劳训练大鼠VEC和VSMC中NF-κB和c-fos均显著性上调表达（P〈0．05）。与有氧运动组比较,疲劳运动大鼠VEC和VSMC中NF-κB和c-fos表达与胸主动脉张开角升高均具有显著性差异（P〈0．05）。表明不同强度运动大鼠主动脉VEC中NF-κB和c-fos表达变化趋势不同,疲劳训练组表达的增加趋势更显著。结论：运动可引起大鼠主动脉VEC和VSMC NF-κB及c-fos表达的变化,且与运动强度关系密切。有氧训练引起的慢性剪切应力变化可使主动脉VEC和VSMC NF-κB加及c-fos显著性下调表达,与VEC和VSMC维持血管功能的稳态有关;疲劳训练可导致壁面摩擦剪切力、周向应力增加,过度的剪切力作用可引起主动脉VEC和VSMC NF-κB及c-fos显著性上调表达,血管张开角显著增加,血管发生非均匀生长,引起血管结构与功能的重塑。     

黄芪对高血压大鼠血管重构中内质网应激反应的影响

目的研究黄芪对高血压大鼠血管重构中内质网应激反应(ERS)的影响,并探讨其血管保护的分子机制。方法将140只大鼠分为对照组、模型组、干预组。采用腹主动脉狭窄术建立高血压大鼠模型,干预组大鼠腹腔注射黄芪注射液8 g/(kg·d)。各组术后1、2、4、6周时采用鼠尾动脉测压法测量大鼠血压,测量血管肌层厚度、Western blot检测CRT和caspase-12的表达、TUNEL法检测血管平滑肌细胞(VSMC)凋亡率。结果模型组术后VSMC形态改变,血压、动脉血管壁肌层厚度和VSMC凋亡率可时间依赖性增大,ERS分子CRT在术后1、2周表达显著升高,4、6周表达降低,而caspase-12分子2周以后表达才升高,且随时间推迟,这种高表达越显著。黄芪干预对比模型组,VSMC形态有一定改善,血压、血管壁肌层厚度和VSMC凋亡率均显著降低,6周时降低幅度最大,同时黄芪能抑制CRT的早期高表达,能抑制caspase-12的高表达,这种抑制作用随着时间的推迟越明显。结论黄芪对高血压大鼠有一定降压作用,可改善血管重构,其机制可能与其调节ERS保护性和促凋亡因子有关。     

体外培养主动脉细胞钙化作用研究

目的：初步探讨主动脉钙化的机制。方法：外殖块法分离培养家兔主动脉平滑肌细胞,进行vonKossa染色以显示钙化,生化法检测细胞内外不溶性钙。放兔法测定培养上清骨钙素的含量。RT-PCR方法检测X型胶原mR-NA的表达。结果：25-羟基胆固醇（A组）和β-甘油磷酸盐（B组）分别培养的传代细胞均可见多个细胞结节,且细胞结节von kossa染色阳性;而对照组（C组）无细胞结节形成,von kossa染色阴性,前二者每孔不溶性钙沉积量,以及培养上清中骨钙素含量明显高于后者,且A、B两组X型胶原mRNA的表达为阳性。结论：25-羟基胆固醇、β-甘油磷酸盐均可促进主动脉中膜细胞发生钙化,主动脉中膜在钙化过程中与成骨细胞相似,骨钙素分泌增多且有X型胶原mRNA的表达。     

EMT促进自发性高血压大鼠主动脉纤维化

目的:探讨内皮间充质转化(endothelial-to-mesenchymal transition,EMT)是否促进自发性高血压大鼠主动脉纤维化及降压治疗对其干预作用。方法:将24只雄性自发性高血压大鼠(spontaneously hypertensive rats,SHR)随机分为SHR组(S组,蒸馏水灌胃)、哌唑嗪组(P组,哌唑嗪5 mg/(Kg×d)灌胃)和氯沙坦组(L组,氯沙坦10 mg/(Kg×d)灌胃),8只雄性WKY大鼠(W组,蒸馏水灌胃)作为对照组,各组分别干预8周后,通过Masson染色检测各组大鼠主动脉纤维化程度,免疫荧光染色及Western Blot检测各组大鼠主动脉I型胶原、CD31及FSP1蛋白表达变化。结果:Western blot及Masson染色显示从L组、P组到S组大鼠主动脉I型胶原含量依次增多,且主动脉壁厚度均显著大于W组(P0.05),但P组与L组大鼠主动脉管壁厚度无统计学差异(P=0.818);免疫荧光染色表明,各组高血压大鼠主动脉均存在FSP1及CD31共表达(FSP+CD31+)细胞,且从L组、P组到S组FSP+CD31+细胞依次增多;Western Blot检测表明,从W组、L组、P组到S组,CD31蛋白表达量逐渐降少,FSP1蛋白表达量依次增加,差异有统计学意义(P0.05)。结论:EMT可能参与了自发性高血压大鼠的主动脉纤维化,氯沙坦和哌唑嗪可能通过抑制EMT减轻主动脉纤维化。     

Gαq/11 和 PDGF依赖性信号转导通路在大鼠主动脉再狭窄中的作用

采用大鼠主动脉球囊内皮剥脱术制备主动脉狭窄模型,观察Gαq/11和PDGF信号转导通路在大鼠主动脉球囊损伤后狭窄时血管平滑肌细胞(VSMC)增殖和迁移中的作用.实验分假手术组、损伤1 d组和损伤14 d组,观察形态学变化,检测血管紧张素转换酶(ACE)活性和主动脉磷脂酶C(PLC)活性,用免疫印迹法测定主动脉血小板源生长因子(PDGF)受体β和Gαq/11蛋白含量.结果显示:损伤1 d,主动脉内皮完全剥脱,VSMC无明显增殖和迁移,内膜无增厚.与假手术组比较,ACE活性增加382.7%(P＜0.01),PDGF受体β表达和PLC活性无明显变化,Gαq/11蛋白含量下降20.0%(P＜0.05).损伤14 d组,主动脉局部有新生内皮出现,中层VSMC大量增殖并向内膜下迁移,内膜显著增厚.ACE活性、PDGF受体β表达和PLC活性分别较假手术组升高420.2%(P＜0.01)、85.0%(P＜0.05)和186.2%(P＜0.05),Gαq/11蛋白表达下降33.1%(P＜0.01).结果提示,PDGF介导的信号转导通路可能是再狭窄时VSMC增殖的重要信号转导机制.     

趋化因子受体CX3CR1调控人主动脉瓣膜间质细胞成骨分化的作用研究

目的:探究趋化因子受体CX3CR1调控人主动脉瓣膜间质细胞成骨分化的作用和机制,为钙化性主动脉瓣膜疾病的早期干预和治疗提供新思路。方法:取非钙化主动脉瓣(3例)和钙化主动脉瓣(5例),免疫组织化学染色检测成骨相关转录因子Runx2、骨桥蛋白OPN和骨钙蛋白OCN的表达;取3例非钙化的主动脉瓣,采用胶原酶连续消化法分离人主动脉瓣膜间质细胞,观察细胞形态及生长状态,并采用细胞免疫荧光进行表型鉴定。对成骨诱导培养的人主动脉瓣膜间质细胞分别过表达和干扰趋化因子受体CX3CR1,平行设置CM组、OM组和negative control+OM组,采用qPCR和Western blot检测Runx2、OPN和OCN的表达,Western blot检测AKT和p-AKT的表达。茜素红S染色评价晚期钙结节形成情况。结果:临床标本显示钙化的主动脉瓣较非钙化的主动脉瓣高表达CX3CR1(P 0. 05);成功分离人主动脉瓣膜间质细胞,α-SMA和Vimentin阳性,vWF阴性。与CM、OM、negative control组比较,CX3CR1+OM组Runx2、OPN和p-AKT表达上调(P 0. 05),且茜素红S染色可见明显钙结节;与CM、OM、negative siRNA control+OM组比较,si CX3CR1+OM组Runx2、OPN和p-AKT表达下调(P 0. 05),且茜素红S染色可见钙结节减少。结论:趋化因子受体CX3CR1可能通过AKT信号通路促进人主动脉瓣膜间质细胞成骨分化。     

人参皂甙Rb1对环磷酰胺所致卵巢早衰大鼠的保护作用

目的探讨人参皂甙Rb1对化疗所致卵巢早衰(Premature ovarian failure,POF)大鼠的Bcl-2及Bax基因表达的影响。方法选用30只2-3月龄具有正常动情周期雌性Wistar大鼠随机分3组,分别为对照组,治疗组和模型组。采用免疫组化和免疫印迹方法分别观察和半定量检测POF大鼠卵巢凋亡调节基因Bcl-2和Bax的蛋白表达情况,同时对比观察人参皂甙Rb1对其表达的影响。结果模型组卵巢颗粒细胞Bax蛋白的平均光密度值较对照组明显增高(P0.05);模型组Bcl-2蛋白的表达量较对照组明显下调(P0.05)。Rb1治疗后Bcl-2蛋白表达明显升高,Bax蛋白表达明显下调(P0.05)。结论 Rb1可能是通过下调bax蛋白水平减少卵巢颗粒细胞的凋亡,对化疗所致卵巢早衰起到治疗的作用,进而延缓卵巢衰老。     

Taurine restores Axl/Gas6 expression in vascular smooth muscle cell calcification model

Our previous studies demonstrated that taurine inhibits osteoblastic differentiation of vascular smooth muscular cells (VSMCs) via the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) signaling pathway, but the underlying mechanism is not elucidated. The tyrosine kinase receptor Axl and its ligand growth arrest-specific protein 6 (Gas6) are expressed in VSMCs. Axl/Gas6 signaling system is known to inhibit VSMCs calcification. We herein showed that taurine partially restored Axl and Gas6 expression in β-glycerophosphate (β-GP)-induced VSMC calcification model. Taurine also induced activation of ERK, but not other two MAPKs including c-jun N-terminal Kinase (JNK) and p38 in VSMCs. Either knockdown of the taurine transporter (TAUT) or treatment with the ERK-specific inhibitor PD98059 blocked the activation of ERK by taurine and abolished taurine-induced Axl/Gas6 expression and calcium deposition reduction in β-GP-induced VSMC calcification model. These results demonstrate for the first time that taurine stimulates expression of Axl and Gas6 via TAUT/ERK signaling pathway in β-GP-induced VSMC calcification model.     

Raloxifene attenuates Gas6 and apoptosis in experimental aortic valve disease in renal failure

Renal failure is associated with aortic valve calcification. Using our rat model of uremia-induced reversible aortic valve calcification, we assessed the role of apoptosis and survival pathways in that disease. We also explored the effects of raloxifene, an estrogen receptor modulator, on valvular calcification. Gene array analysis was performed in aortic valves obtained from three groups of rats (n = 7 rats/group): calcified valves obtained from rats fed with uremic diet, valves after calcification resolution following diet cessation, and control. In addition, four groups of rats (n = 10 rats/group) were used to evaluate the effect of raloxifene in aortic valve calcification: three groups as mentioned above and a fourth group fed with the uremic diet that also received daily raloxifene. Evaluation included imaging, histology, and antigen expression analysis. Gene array results showed that the majority of the altered expressed genes were in diet group valves. Most apoptosis-related genes were changed in a proapoptotic direction in calcified valves. Apoptosis and decreases in several survival pathways were confirmed in calcified valves. Resolution of aortic valve calcification was accompanied by decreased apoptosis and upregulation of survival pathways. Imaging and histology demonstrated that raloxifene significantly decreased aortic valve calcification. In conclusion, downregulation of several survival pathways and apoptosis are involved in the pathogenesis of aortic valve calcification. The beneficial effect of raloxifene in valve calcification is related to apoptosis modulation. This novel observation is important for developing remedies for aortic valve calcification in patients with renal failure.     

α-Lipoic acid attenuates vascular calcification via reversal of mitochondrial function and restoration of Gas6/Axl/Akt survival pathway

Vascular calcification is prevalent in patients with chronic kidney disease and leads to increased cardiovascular morbidity and mortality. Although several reports have implicated mitochondrial dysfunction in cardiovascular disease and chronic kidney disease, little is known about the potential role of mitochondrial dysfunction in the process of vascular calcification. This study investigated the effect of α-lipoic acid (ALA), a naturally occurring antioxidant that improves mitochondrial function, on vascular calcification in vitro and in vivo. Calcifying vascular smooth muscle cells (VSMCs) treated with inorganic phosphate (Pi) exhibited mitochondrial dysfunction, as demonstrated by decreased mitochondrial membrane potential and ATP production, the disruption of mitochondrial structural integrity and concurrently increased production of reactive oxygen species. These Pi-induced functional and structural mitochondrial defects were accompanied by mitochondria-dependent apoptotic events, including release of cytochrome c from the mitochondria into the cytosol, subsequent activation of caspase-9 and -3, and chromosomal DNA fragmentation. Intriguingly, ALA blocked the Pi-induced VSMC apoptosis and calcification by recovery of mitochondrial function and intracellular redox status. Moreover, ALA inhibited Pi-induced down-regulation of cell survival signals through the binding of growth arrest-specific gene 6 (Gas6) to its cognate receptor Axl and subsequent Akt activation, resulting in increased survival and decreased apoptosis. Finally, ALA significantly ameliorated vitamin D(3) -induced aortic calcification and mitochondrial damage in mice. Collectively, the findings suggest ALA attenuates vascular calcification by inhibiting VSMC apoptosis through two distinct mechanisms; preservation of mitochondrial function via its antioxidant potential and restoration of the Gas6/Axl/Akt survival pathway.     

MiR155 modulates vascular calcification by regulating Akt-FOXO3a signalling and apoptosis in vascular smooth muscle cells

microRNA-155 (miR155) is pro-atherogenic; however, its role in vascular calcification is unknown. In this study, we aim to examine whether miR155 regulates vascular calcification and to understand the underlying mechanism. Quantitative real-time PCR showed that miR155 is highly expressed in human calcific carotid tissue and positively correlated with the expression of osteogenic genes. Wound-healing assay and TUNEL staining showed deletion of miR155 inhibited vascular smooth muscle cell (VSMC) migration and apoptosis. miR155 deficiency attenuated calcification of cultured mouse VSMCs and aortic rings induced by calcification medium, whereas miR155 overexpression promoted VSMC calcification. Compared with wild-type mice, miR155^−/− mice showed significant resistance to vitamin D3 induced vascular calcification. Protein analysis showed that miR155 deficiency alleviated the reduction of Rictor, increased phosphorylation of Akt at S473 and accelerated phosphorylation and degradation of FOXO3a in cultured VSMCs and in the aortas of vitamin D3-treated mice. A PI3K inhibitor that suppresses Akt phosphorylation increased, whereas a pan-caspase inhibitor that suppresses apoptosis reduced VSMC calcification; and both inhibitors diminished the protective effects of miR155 deficiency on VSMC calcification. In conclusion, miR155 deficiency attenuates vascular calcification by increasing Akt phosphorylation and FOXO3a degradation, and thus reducing VSMC apoptosis induced by calcification medium.     

Identification of a protective role for protein phosphatase 1cgamma1 against oxidative stress-induced vascular smooth muscle cell apoptosis

The development of therapeutic strategies to inhibit reactive oxygen species (ROS)-mediated damage in blood vessels has been limited by a lack of specific targets for intervention. Targeting ROS-mediated events in the vessel wall is of interest, because ROS play important roles throughout atherogenesis. In early atherosclerosis, ROS stimulate vascular smooth muscle cell (VSMC) growth, whereas in late stages of lesion development, ROS induce VSMC apoptosis, causing atherosclerotic plaque instability. To identify putative protective genes against oxidative stress, mouse aortic VSMC were infected with a retroviral human heart cDNA expression library, and apoptosis was induced in virus-infected cells by 2,3-dimethoxy-1,4-naphthoquinone (DMNQ) treatment. A total of 17 different, complete cDNAs were identified from the DMNQ-resistant VSMC clones by PCR amplification and sequencing. The cDNA encoding PP1cgamma1 (catalytic subunit of protein phosphatase 1) was present in several independent DMNQ-resistant VSMC clones. DMNQ increased mitochondrial ROS production, caspase-3/7 activity, DNA fragmentation, and decreased mitochondrial transmembrane potential in VSMC while decreasing PP1cgamma1 activity and expression. Depletion of PP1cgamma1 expression by short hairpin RNA significantly enhanced basal as well as DMNQ-induced VSMC apoptosis. PP1cgamma1 overexpression abrogated DMNQ-induced JNK1 activity, p53 Ser(15) phosphorylation, and Bax expression and protected VSMC against DMNQ-induced apoptosis. In addition, PP1cgamma1 overexpression attenuated DMNQ-induced caspase-3/7 activation and DNA fragmentation. Inhibition of p53 protein expression using small interfering RNA abrogated DMNQ-induced Bax expression and significantly attenuated VSMC apoptosis. Together, these data indicate that PP1cgamma1 overexpression promotes VSMC survival by interfering with JNK1 and p53 phosphorylation cascades involved in apoptosis.     

Cav1.2在顺铂诱导C57BL/6J小鼠耳蜗螺旋神经元凋亡中的作用*

目的: 探究顺铂(CDDP)诱导C57BL/6J小鼠耳蜗螺旋神经元(SGNs)凋亡过程中Cav1.2的作用及其可能的机制。方法: 动物实验:选取8周龄雄性C57BL/6J小鼠分为以下两组(10只/组):生理盐水组(Control组)和顺铂给药组(Cisplatin组)。Control组每天腹腔注射生理盐水,Cisplatin组每周期前4 d以3 mg/kg的剂量进行顺铂腹腔注射,后10 d每日注射生理盐水,重复三个周期。给药结束后,听性脑干反应(ABR) 检测小鼠听力阈值变化; 小鼠内眦采血,并断颈取耳蜗,超氧化物歧化酶(SOD)以及丙二醛(MDA)试剂盒检测血清及耳蜗组织的SOD活性和MDA含量;免疫印迹法(Western blot)检测耳蜗组织相关凋亡蛋白表达;苏木精-伊红HE染色观察小鼠耳蜗螺旋神经节形态学变化; TUNEL 染色观察小鼠耳蜗SGNs凋亡情况;免疫荧光观察耳蜗SGNs上Cav1.2的分布和表达。细胞实验:原代培养SGNs,根据CCK8选择顺铂5 μmol/L干预12 h并随机分为:对照组(Control)、溶剂组(DMSO)、Cav1.2阻断剂组(N)、顺铂组(Cisplatin)、顺铂与Cav1.2阻断剂共同孵育组(Cisplatin+N)。Western blot检测Cav1.2蛋白表达;Hoechst33342染色观察各组SGNs凋亡情况,流式细胞术检测各组SGNs凋亡率,Western blot检测相关凋亡蛋白的表达,CA²⁺探针检测细胞内钙离子浓度变化,线粒体膜电位检测试剂盒(JC-1)检测膜电位变化,线粒体超氧化物指示剂(MitoSOX^TM-red)检测线粒体释放ROS情况。结果: 动物实验:与Control组相比,Cisplatin组小鼠听力阈值升高(P<0.01), 血清及耳蜗组织MDA含量、耳蜗组织凋亡蛋白 Cleaved-caspase-3、Bax 蛋白水平和TUNEL阳性率、Cav1.2蛋白表达水平等均明显升高(P<0.05, P<0.01);血清及耳蜗组织SOD活性、耳蜗组织抗凋亡蛋白 Bcl-2 蛋白水平和SGCs密度均明显降低(P<0.05,P<0.01)。细胞实验:与Control组相比,Cisplatin组的Cav1.2表达、细胞凋亡率、Cleaved-caspase-3、Bax蛋白水平、细胞内钙离子浓度以及ROS释放均明显增加(P<0.05,P<0.01);而细胞的Bcl-2蛋白水平和线粒体膜电位则明显降低(P<0.01);Cav1.2阻断剂可部分逆转上述改变(P<0.05)。 结论: 顺铂可能通过上调Cav1.2促进钙内流,进而使线粒体ROS增多,引起SGNs氧化应激损伤从而诱导线粒体途径的细胞凋亡。