药物保健与健康系统杂志
开放获取

抽象的

微流体混合制备用于 siRNA 递送的有毒纳米粒子的过程

穆罕默德·奥贝德

RNA 干扰涉及通过掺入短干扰 RNA (siRNA) 来降解目标信使 RNA [1]。RNA 干扰 (RNAi) 是最流行的生物工具，双链 RNA (dsRNA) 通过靶向互补 mRNA 进行降解来促进基因沉默，是疾病普遍治疗方法的一项巨大创新，并改变了研究人员研究基因功能的方式。siRNA 扩展了有关内源性 RNA 阻抗的亚原子成分的信息，已被开发为富有想象力的核酸腐蚀药物，用于治疗恶性生长等致命疾病。

 

小干扰 RNA (siRNA) 被误导性地组合了 19-23 个核苷酸长的双倍废弃 RNA 原子。它们通常用于亚原子科学，以暂时平息阴谋的质量。它们在官方对其目标转录本依赖于连续互补性后激发 RNAi 反应。它们已被适当地用于考虑不同致癌 lncRNA 通过容量损失而产生的影响。

 

The siRNA has given new chances to the improvement of inventive medication to treat beforehand serious illnesses, for example, malignant growth. It is of natural intensity since it misuses the endogenous RNAi pathway, permits explicit decrease of ailment related qualities, and is appropriate to any quality with a correlative arrangement. For the basis of siRNA-intervened quality treatment, hereditary nature of disease offers strong help. Truth be told, various siRNAs have been intended to target predominant oncogenes, viral oncogenes associated with carcinogenesis, or mal practically directed oncogenes.

 

Mechanism:

 

The initial step of RNAi includes handling and cleavage of longer twofold abandoned RNA into siRNAs, by and large bearing a 2 nucleotide overhang on the 3′ end of each strand. The protein answerable for this handling is a RNase III-like catalyst named Dicer. At the point when shaped, siRNAs are limited by a multiprotein part complex alluded to as RISC (RNAinduced hushing complex). Inside the RISC complex, siRNA strands are isolated and the strand with the more steady 5′-end is ordinarily incorporated to the dynamic RISC complex. The antisense single-abandoned siRNA part at that point controls and adjusts the RISC complex on the objective mRNA and through the activity of synergist RISC protein, an individual from the argonaute family (Ago2), mRNA is severed.

 

Methods:

 

NISV were set up by microfluidic blending which is an as of late created technique used to get ready lipid based nanoparticles and results in the creation of little vesicles with effective exemplification of a restorative operator. To get ready NISV, explicit volumes from each stock arrangement of the NISV parts were combined to set up the lipid stage. The lipid stage was infused into the main gulf and the fluid stage into the second bay of the microfluidic micromixer, with the blending temperature set at 50°C. The stream rate proportions (FRR) between the watery and natural stage was set at 3:1 and the all out stream rates (TFR) of the two stages was set at 12 ml/min. This takes into consideration quick blending between the two stages at high stream rates and at a temperature over the stage progress of the lipids. Scatterings were then gathered from the outlet stream and quickly weakened so as to lessen the last ethanol content in the planning to 6.25% (v/v). Cytotoxicity assessment of NISV were completed on non-little lung malignancy cells (A549) and mouse melanoma cells (B16-F10-LUC). siRNA focusing on green fluorescent protein (GFP) in copGFP-A549 cells, or luciferase in B16-F10-LUC cells were typified in NISV. Restraint of GFP articulation by against GFP siRNA (siGFP) conveyed utilizing NISV was assessed by stream cytometry, polymerase chain response, and Western smudging. Naked BALB/c mice vaccinated with B16-F10-LUC cells that prompt melanoma communicating luciferase was utilized to survey the NISV capacity to convey siRNA in vivo.

 

Results:

 

Cytotoxicity considers demonstrated that NISV were not harmful at or beneath 40 µg/ml. NISV definitions had high siRNA exemplification effectiveness. Fluorescent magnifying lens and stream cytometry considers demonstrated high cell take-up by the cells contrasted with bare siRNA, which was not taken up by the cells. NISV had the option to convey siGFP to the cells and altogether smother GFP articulation. These outcomes were affirmed by transfecting the luciferase creating B16-F10-LUC cells with against luciferase siRNA (siLUC). Estimating the degree of luciferase articulation after siLUC transfections utilizing a luciferase protein measure framework effectively exhibited the concealment of luciferase articulation. NISV were then utilized in vivo tests utilizing naked BALB/c mice. After intra-tumoural infusion, siLUC was conveyed to the cells and stifled luciferase articulation at an essentially more elevated level than mice rewarded with exposed siLUC. These in vivo outcomes affirm the capacity of NISV to effectively convey siRNA into the cytoplasm of the objective cells and stifle the objective protein.

 

结论：

 

NISV 已被广泛展示，只是因为它可以用作 siRNA 的传输框架。这些结果表明，NISV 可用于克服基于 siRNA 的治疗中的障碍，例如稳定性低和细胞吸收差。