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丝胶蛋白-壳聚糖共轭银纳米颗粒通过调节转移生物标志物、抑制发育异常和增强抗氧化潜力,对1,2-二甲基肼诱导的小鼠结肠癌起到保护作用。

Sericin-chitosan conjugated silver nanoparticles protect against 1,2-dimethylhydrazine-induced colon cancer in mice by regulating metastatic biomarkers, prohibiting dysplasia and enhancing antioxidant potential.

作者信息

Ijaz Farah, Ali Shaukat, Pervaiz Asim, Khan Khushbukhat, Afsar Tayyaba, Aldisi Dara, Amor Houda, Razak Suhail

机构信息

Medical Toxicology and Biochemistry Laboratory, Department of Zoology, Government College University, Lahore 54000, Pakistan.

Medical Toxicology and Biochemistry Laboratory, Department of Zoology, Government College University, Lahore 54000, Pakistan.

出版信息

Int J Biol Macromol. 2025 Sep;321(Pt 3):146478. doi: 10.1016/j.ijbiomac.2025.146478. Epub 2025 Jul 31.

Abstract

Colon cancer is reported as third most prevalent malignancy worldwide, while sericin being an antioxidant, is now used in biomedical applications due to its biochemical characteristics and has shown potential efficacy to treat colon cancer. Sericin was isolated by the degumming process followed by the characterization by using FTIR, UV, XRD, and SEM techniques to confirm the successful synthesis of SAgNPs and SChiAgNPs. The male Balb C mice were then divided into 13 groups. Group 1: Control, Group 2: DMH (20 mg/kg) (injected (IP) thrice a week for 14 weeks). Groups 3,4,5: Sericin (S) (100 mg/kg), Sericin silver nanoparticles (SAgNPs) (100 mg/kg), and Sericin Chitosan silver nanoparticles (SChiAgNPs) (100 mg/kg) were given orally for 14 weeks respectively. Groups 6,7,8,9 were considered as preventive groups and were given DMH (IP) + 5-FU (IP), DMH(IP) + S (orally), DMH (IP) + SAgNPs (orally), DMH (IP) + SChiAgNPs (orally) respectively for the period of 14 weeks Groups 10,11,12,13 were considered as treatment groups and were given 5-FU (5 mg/kg) (IP), (S) (100 mg/kg) (orally), (SAgNPs) (100 mg/kg) (orally), (SChiAgNPs) (100 mg/kg) (orally) for a period of first 7 weeks after 7 weeks of DMH administration (IP). After 14 weeks period study, blood samples and colon tissue were used for the analysis of biochemical markers i.e., CEA, CA19-9, TIMP-1, and IL-6, IL-8, IL-27, SOD, CAT, GR, GSH, GST, MDA and MMP-7 via ELISA and histopathological analysis. The UV absorption peaks obtained at 435 and 463 nm indicated the formation of SAgNPs and SChiAgNPs formation respectively. FTIR spectra peaks obtained, indicate NH stretching of primary and secondary amine group), (NH stretching of amine salt) (N=C=S stretching of thiocyanate compound), (CC stretching of alkene), (NO stretching of nitro compound), (SO stretching of sulfonyl chloride), (CN stretching of amine) and (C-O-O stretching) for sericin, SAgNPs, and SChiAgNPs, confirming, the presence of theses functional groups. The XRD pattern revealed that SAgNPs and SChiAgNPs had structure crystalline structures. EDX characterization peaks for SAgNPs indicated the presence of silver along with other elements including; calcium, oxygen carbon, while EDX characterization peaks for SChiAgNPs indicated the presence of silver along with other elements including; oxygen, carbon, sodium, phosphorus, Sulphur and chlorine. SEM analysis showed that SAgNPs are of spherical shape, while the SChiAgNPs displayed the rectangular shape. The results for biomarker analysis indicated significantly elevated levels of CEA, CA19-9, TIMP-1, IL-6, IL-8, IL-27, MDA, and MMP-7 in DMH treated group (p ≤ 0.001) which were decreased significantly in SChiAg(T) (p ≤ 0.001). In contrast, levels of SOD, GR, GSH, CAT and GST were reduced significantly in DMH treated group, which were increased significantly in SChiAg(T) (p ≤ 0.001). The histopathological analysis of proximal and distal parts of colon tissue of the DMH-treated group showed low-grade dysplasia (LGD), and high-grade dysplasia (HGD) while SChiAgNPs improved the histopathological changes induced by DMH. The findings suggest that Sericin Chitosan conjugated silver nanoparticles showed their efficacy against DMH-induced colon cancer, making a potential future in the anticancer research field.

摘要

据报道,结肠癌是全球第三大常见恶性肿瘤,而丝胶蛋白作为一种抗氧化剂,由于其生化特性,目前已被用于生物医学应用,并显示出治疗结肠癌的潜在疗效。通过脱胶工艺分离丝胶蛋白,然后使用傅里叶变换红外光谱(FTIR)、紫外可见光谱(UV)、X射线衍射(XRD)和扫描电子显微镜(SEM)技术进行表征,以确认成功合成了丝胶银纳米颗粒(SAgNPs)和丝胶壳聚糖银纳米颗粒(SChiAgNPs)。然后将雄性Balb C小鼠分为13组。第1组:对照组;第2组:二甲基肼(DMH,20 mg/kg)(每周腹腔注射3次,共14周)。第3、4、5组:分别口服丝胶蛋白(S,100 mg/kg)、丝胶银纳米颗粒(SAgNPs,100 mg/kg)和丝胶壳聚糖银纳米颗粒(SChiAgNPs,100 mg/kg),持续14周。第6、7、8、9组被视为预防组,分别给予DMH(腹腔注射)+5-氟尿嘧啶(5-FU,腹腔注射)、DMH(腹腔注射)+丝胶蛋白(口服)、DMH(腹腔注射)+丝胶银纳米颗粒(口服)、DMH(腹腔注射)+丝胶壳聚糖银纳米颗粒(口服),持续14周。第10、11、12、13组被视为治疗组,在DMH腹腔注射7周后,前7周分别给予5-氟尿嘧啶(5 mg/kg,腹腔注射)、丝胶蛋白(100 mg/kg,口服)、丝胶银纳米颗粒(100 mg/kg,口服)、丝胶壳聚糖银纳米颗粒(100 mg/kg,口服)。经过14周的研究期后,采集血液样本和结肠组织,通过酶联免疫吸附测定(ELISA)和组织病理学分析,分析生化标志物,即癌胚抗原(CEA)、糖类抗原19-9(CA19-9)、基质金属蛋白酶组织抑制因子-1(TIMP-1)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、白细胞介素-27(IL-27)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽还原酶(GR)、谷胱甘肽(GSH)、谷胱甘肽S-转移酶(GST)、丙二醛(MDA)和基质金属蛋白酶-7(MMP-7)。在435和463 nm处获得的紫外吸收峰分别表明形成了SAgNPs和SChiAgNPs。获得的FTIR光谱峰表明,丝胶蛋白、SAgNPs和SChiAgNPs存在伯胺和仲胺基团的NH伸缩振动、胺盐NH伸缩振动、硫氰酸盐化合物的N=C=S伸缩振动、烯烃的CC伸缩振动、硝基化合物的NO伸缩振动、磺酰氯的SO伸缩振动、胺的CN伸缩振动和(C-O-O)伸缩振动,证实了这些官能团的存在。XRD图谱显示SAgNPs和SChiAgNPs具有晶体结构。SAgNPs的能谱分析(EDX)表征峰表明存在银以及其他元素,包括钙、氧、碳;而SChiAgNPs的EDX表征峰表明存在银以及其他元素,包括氧、碳、钠、磷、硫和氯。SEM分析表明,SAgNPs呈球形,而SChiAgNPs呈矩形。生物标志物分析结果表明,DMH处理组的CEA、CA19-9、TIMP-1、IL-6、IL-8、IL-27、MDA和MMP-7水平显著升高(p≤0.001),而丝胶壳聚糖银纳米颗粒治疗组(SChiAg(T))则显著降低(p≤0.001)。相反,DMH处理组的SOD、GR、GSH、CAT和GST水平显著降低,而丝胶壳聚糖银纳米颗粒治疗组则显著升高(p≤0.001)。DMH处理组结肠组织近端和远端的组织病理学分析显示为低级别发育异常(LGD)和高级别发育异常(HGD),而丝胶壳聚糖银纳米颗粒改善了DMH诱导的组织病理学变化。研究结果表明,丝胶壳聚糖偶联银纳米颗粒对DMH诱导的结肠癌具有疗效,在抗癌研究领域具有潜在的应用前景。

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