C-Phycocyanin Attenuates Cisplatin-Induced Nephrotoxicity in Mice

Although cisplatin is a highly effective antineoplastic agent, nephrotoxicity is its major clinical problem. Recently, it was reported that Spirulina, a blue-green algae, has potent antioxidant properties. The aim of this study was to establish the possible protective role of C-phycocyanin (PC), one of the active ingredients of Spirulina, against cisplatin-induced nephrotoxicity. This study was carried out using human kidney-2 (HK-2) cells and male C57BL6 mice. Cells and mice were divided into four groups; untreated control group, PC-treated control group, cisplatin-treated group, and PC plus cisplatin-treated group. The molecular, functional, and structural parameters were measured. PC significantly attenuated blood urea nitrogen, serum creatinine, renal histological damages, and apoptotic cell death in cisplatin-treated mice. The cisplatin-induced cell death was significantly attenuated in cells pretreated with PC. PC also significantly attenuated the elevation of p-ERK, p-JNK, and p-p38 induced by cisplatin treatment. The expression of Bax, caspase-9, and caspase-3 in cisplatin-treated cells were also decreased by PC treatment. In conclusion, PC ameliorates cisplatin-induced nephrotoxicity and, at least in part, suppression of p-ERK, p-JNK, p-p38, Bax, caspase-9, and caspase-3 may be involved in this mechanism.     

藻蓝蛋白联合全反式维甲酸诱导HeLa细胞凋亡的分子机制研究

目的 探讨全反式维甲酸和藻蓝蛋白联合用药对HeLa细胞生长的影响,并分析两者联合用药诱导细胞凋亡可能的分子机制。方法 MTT法检测全反式维甲酸和藻蓝蛋白单独及联合用药对HeLa细胞生长的影响,TUNEL法检测单独及联合用药HeLa细胞凋亡情况,免疫组化法检测Bcl-2基因的表达情况,Westren blot法检测Caspase-3的表达情况。结果 全反式维甲酸和藻蓝蛋白均具有抑制HeLa细胞生长的作用,当达到相同的抑制率时,联合藻蓝蛋白使用可以显著降低全反式维甲酸的使用剂量从而达到降低毒副作用的目的。2种药物联合用药处于中高抑制率时,2种药物为协同作用。两者联合用药可以显著下调Bcl-2和增加Caspase-3的表达水平从而引发细胞凋亡。结论 2种药物单独使用均能抑制HeLa细胞的生长,联合用药时该抑制作用更为显著。全反式维甲酸和藻蓝蛋白联合用药诱导HeLa细胞凋亡的分子机制可能是通过抑制bcl-2基因的表达、促进Caspase-3蛋白的表达,从而促进凋亡信号的转导最终导致细胞死亡的。 关键词：全反式维甲酸;藻蓝蛋白;HeLa细胞;凋亡;增殖抑制     

藻蓝蛋白-光动力疗法治疗小鼠HeLa细胞瘤的免疫和凋亡机制研究

目的探讨藻蓝蛋白介导的光动力疗法治疗小鼠HeLa细胞瘤的免疫和凋亡机制。方法将HeLa细胞接种于小鼠肋缘皮下脾区构建荷瘤小鼠模型。实验分成3组：对照组、激光照射组、光动力治疗组（PDT组）。PDT组：肿瘤局部皮下注射藻蓝蛋白液0.25ml（约2g）2h后以He-Ne激光照射。实验第7d测瘤块重量,取胸腺、脾脏检测NK细胞活性和T细胞增殖活性。取瘤块制成石蜡包埋切片,采用原位核酸杂交技术、免疫组织化学技术检测肿瘤细胞内CD44、P53、NFκB、Fas基因的表达。结果与对照组相比,激光照射组NK细胞和免疫T细胞的增殖活性有所增强,肿瘤细胞内抗凋亡基因（Fas）表达量显著增多,而瘤块的重量、肿瘤形成率和抗凋亡基因（P53、NFκB、CD44）明显减少。以上各项指标PDT组与激光组比较,差异亦具有显著或非常显著意义（P〉0.05或P〈0.01）。结论藻蓝蛋白介导的光动力疗法通过增强机体的免疫力同时启动HeLa细胞内的凋亡信号转导通路诱导细胞凋亡,从而达到杀死肿瘤的目的。     

藻蓝蛋白抗肿瘤的应用现状

近年来,在海洋生物中寻找高效、低毒、不良反应小的抗肿瘤药物已引起了国内外学者的重视。藻蓝蛋白（C-phycocyanin, C-PC）对多种肿瘤细胞具有毒副作用,可以抑制肿瘤细胞的生长,促进肿瘤细胞的凋亡。联合其他药物可以提高藻蓝蛋白的抗肿瘤活性。现对藻蓝蛋白抗肿瘤作用的研究进展进行总结,为新型天然药物的研发提供依据。     

新型羧甲基壳聚糖包裹C-藻蓝蛋白负载CD59配体的靶向纳米微球的制备及其对HeLa细胞的杀伤作用

目的：制备一种新型的C-藻蓝蛋白（C-phycocyanin,C-PC）靶向纳米微球（nanometer microspheres,NPs）,并探究其对HeLa细胞的靶向治疗作用。方法：采用离子交联法制备靶向羧甲基壳聚糖（carboxymethyl chitosan,CMC ）纳米载药颗粒C-PC/CMC-NPs;响应面优化法筛选出最佳制备条件,并用透射电镜、激光粒度仪观察纳米载药颗粒的表征;MTT法检测靶向纳米颗粒C-PC/CMC-CD59sp-NPs对人宫颈癌HeLa细胞生长的影响;溶血实验检测C-PC/CMC-CD59sp-NPs的组织相容性及安全性;Western blotting和免疫组化法分别检测C-PC/CMC-CD59sp-NPs 对HeLa细胞内活化的Caspase-3/PARP蛋白和Bcl-2蛋白表达的影响。结果： 成功制备C-PC/CMC纳米微球,透射电镜观察到纳米微球呈现出规则的球形,分散均匀,其粒径约为160 nm。最佳制备条件：羧甲基壳聚糖浓度为2.0 mg/ml,CaCl2浓度为1.0 mg/ml,粒径约为120 nm的球形,包封率为(62±5)%;CMC与C-PC投药量为3∶1,载药量为(20±3)%;C-PC/CMC-NPs体外表现出缓释特征,在120 h内pH=5.4和74条件下,释放率分别达到80%和60%;同时未出现溶血现象。C-PC/CMC-CD59sp-NPs对HeLa细胞增殖有明显的抑制作用,促进活化的Caspase-3/PARP蛋白的表达,抑制Bcl-2蛋白的表达（均P＜0.05）。结论：新型靶向纳米药物C-PC/CMC-CD59sp-NPs能在体外抑制HeLa细胞的生长,诱导其凋亡。为海洋药物的研发提供了新的思路,对靶向纳米药物的进一步研究提供理论基础。     

Protection against cisplatin-induced nephrotoxicity by Spirulina in rats

Purpose: Cisplatin (CP)-induced nephrotoxicity is associated with the increased generation of reactive oxygen metabolites and lipid peroxidation in kidney, caused by the decreased levels of antioxidants and antioxidant enzymes. The purpose of this study was to evaluate the role of Spirulina, blue–green alga with antioxidant properties, in the protection of cisplatin-induced nephrotoxicity in rat. Methods: Rats were treated with CP (6 mg/kg bw, single dose, intraperitoneally). Spirulina (1,000 mg/kg) was administered orally for 8 days and CP treatment was given on day 4. Nephrotoxicity was assessed, 6 days after the CP treatment, by measuring plasma urea, creatinine, urinary N-acetyl-(d-glucose-aminidase) (β-NAG) and histopathology of kidney. Results: Rats treated with CP showed marked nephrotoxicity as evidenced from the significant elevation in plasma urea, creatinine and urinary β-NAG. Histological assessment revealed marked proximal tubular necrosis and extensive epithelial vacuolization in the kidney of CP-treated rats. Superoxide dismutase, catalase and glutathione peroxidase were decreased and lipid peroxidation was increased in kidney tissue. Pretreatment with Spirulina protected the rats from CP-induced nephrotoxicity. The rise in plasma urea, creatinine, urinary β-NAG, plasma and kidney tissue MDA and histomorphological changes were significantly attenuated by Spirulina. In vitro studies using human ovarian cancer cells revealed that Spirulina did not interfere with the cytotoxic effects of CP on tumor cells. Conclusions: In summary, Spirulina significantly protected the CP-induced nephrotoxicity through its antioxidant properties.     

CPC/CMC-CD55sp 纳米微球对宫颈癌Caski 细胞的靶向杀伤作用

目的:制备一种新型C-藻蓝蛋白（C-phycocyanin,CPC）/羧甲基壳聚糖-CD55 配体肽（carboxymethyl chitosan-CD55-specific ligand peptide, CMC-CD55sp）(CPC/CMC-CD55sp)纳米微球,并探究其对宫颈癌Caski 细胞的靶向治疗作用。方法：采用离子交联法制备新型CPC/CMC-CD55sp 纳米微球,通过透射电镜（DLS）和红外光谱仪（FTIR）观察纳米微球的表征,Western blotting和流式细胞术检测CD55 在Caski 细胞和成纤维（L-929）细胞表面的表达情况,CCK-8 法检测纳米微球对Caski 细胞增殖的影响,流式细胞术和荧光显微镜检测纳米微球被细胞摄取情况,Western blotting 和流式细胞术检测纳米微球对Caski 细胞凋亡相关信号蛋白和凋亡率的影响,溶血试验测定药物的生物安全性。结果：成功制备新型CPC/CMC-CD55sp 纳米微球,其形态良好、直径分布均匀,CD55 在宫颈癌细胞Caski 表面高表达而在小鼠L-929 细胞表面低表达（P<0.01）。纳米微球能靶向、高效地到达Caski细胞并被摄入细胞;其在人外周血中溶血作用微弱,且在安全范围;其对Caski 细胞的增殖具有明显的抑制作用并且能够诱导Caski 细胞凋亡（P<0.05 或P<0.01）。结论：新型CPC/CMC-CD55sp 纳米微球能够靶向抑制宫颈癌Caski 细胞的增殖并诱导其凋亡,具有较好的安全性,为海洋抗肿瘤药物的研发提供了新思路。     

藻蓝蛋白介导光动力疗法在乳腺癌治疗中的机制研究

目的探讨藻蓝蛋白介导的光动力学疗法在乳腺癌治疗中的机制。方法将MCF-7细胞接种于小鼠肋缘皮下脾区构建乳腺癌小鼠模型。小鼠分成4组:对照组、He-Ne激光照射组、藻蓝蛋白处理组、光动力学治疗组(PDT组)。10d后检测瘤块重量,NK细胞活性和T细胞增殖活性。取瘤块制成石蜡包埋切片,采用原位核酸杂交技术、免疫组织化学技术检测组织细胞内凋亡相关蛋白的表达。体外培养的MCF-7细胞也相应分成4组,通过MTT法、电镜和免疫荧光技术检测细胞增殖活性、细胞形态、细胞色素C表达量的变化。结果与对照组相比,激光照射组各检测指标均无明显差异,而藻蓝蛋白处理组中NK细胞和T细胞的增殖活性有所增强,肿瘤组织细胞内抗凋亡蛋白(Fas)表达量明显增多,而瘤块的重量、肿瘤形成率和抗凋亡蛋白(p53、NF-κB、CD44)明显减少,如果藻蓝蛋白结合激光治疗发现各检测指标与对照组比较差异更为明显。体外试验证实藻蓝蛋白能抑制MCF-7细胞的增殖,促进细胞色素C的释放,电镜下细胞呈现典型的凋亡形态,用光动力学方法处理后效果更为明显。结论藻蓝蛋白可以作为一种光敏剂,其介导的光动力学疗法通过增强机体的免疫力同时启动乳腺癌细胞内的凋亡信号转导通路诱导细胞凋亡,从而达到杀死肿瘤的目的。     

钝顶螺旋藻藻蓝蛋白抗百草枯诱导大鼠肺纤维化的研究

目的 探讨钝顶螺旋藻(spirulina platensis)藻蓝蛋白(C-phycoeyanin,C-PC)对百草枯(paraquat,PQ)染毒诱导大鼠肺纤维化的治疗作用.方法 选取健康Wistar大鼠90只,随机分为正常对照组、模型组(PQ组)和藻蓝蛋白治疗组(C-PC组),每组30只.PQ组、C-PC组以PQ(50 mg/kg)一次灌胃染毒造模.造模后,正常对照组、PQ组大鼠每天给予生理盐水(1 ml/100 g)灌胃,C-PC组每天给予C-PC (50 mg/kg)灌胃,并在第1、3、7、14、28天各组随机取6只大鼠右肺下叶肺组织匀浆测羟脯氨酸(HYP)、丙二醛(MDA)含量、超氧化物歧化酶(SOD)活力;取左肺部分组织行HE染色、Masson染色进行病理观察,并用免疫组化方法观察肺组织中转化生长因子-β1(TGF-β1)、核因子-κB亚基p65(NF-κB p65)及肿瘤坏死因子-α(TNF-α)的表达.结果 各观察时点C-PC组大鼠肺组织中HYP和14、28 d MDA含量明显低于PQ组,差异有统计学意义(P＜0.05,P＜0.01).1、7、14、28 d时C-PC组大鼠肺组织中SOD活力明显高于PQ组,差异有统计学意义(P＜0.05,P＜0.01).PQ组和C-PC组肺组织中TGF-β1的表达及NF-κB p65和TNF-α表达明显高于正常对照组,C-PC组肺组织的TGF-β1表达及NF-κB p65和TNF-α表达明显低于PQ组,差异均有统计学意义(P＜0.05,P＜0.01).病理学观察显示C-PC能减轻PQ染毒大鼠肺泡炎和肺纤维化程度.结论 C-PC对PQ诱导的大鼠肺泡炎及肺纤维化有抑制作用.     

Spirulina maxima and its protein extract protect against hydroxyurea-teratogenic insult in mice

Congenital malformations are one of the major causes of child mortality all over the world. In order to prevent them it is necessary to find substances that act as anti-teratogenic agents. In this study hydroxyurea (HU), an antineoplastic and teratogenic drug, was administered to pregnant mice because one of its major mechanisms of teratogenesis is the production of reactive oxygen species (ROS). The aim of this work was to determine if Spirulina maxima (SP) and its aqueous protein extract could protect against HU-teratogenic insult in mouse embryos. SP has been used for a long time because of its nutritional and pharmacological properties. The antioxidant activity, one of the most important, is related to the protein extract due to its content of phycobiliproteins. It was observed that neither SP nor its extract provoked teratogenic effects at any dose tested and even increased vitelline yolk sac circulation. Dams exposed to HU (30 mg/kg, i.p.) presented embryos with multiple alterations in their development. Groups treated with SP or its extract, before and after HU exposure, showed a protector effect in a dose-dependent manner. TBARS test confirmed that the protection effect was related to the antioxidant activity of both SP and its extract.