取向性生物导电型材料的制备及对神经再生的影响*

龚佳欢1**,韦自卫2,石嘉琪2,牛长梅2,赵亚红1,杨宇民1

1南通大学神经再生重点实验室;2南通大学医学院,江苏226001

[ ] 目的:通过不同方法研制出取向性静电纺丝复合材料,确定材料与神经再生之间的构效关系,为理想的神经再生植入装置的设计提供扎实的理论基础,为周围神经缺损的修复提供更有效的方法。方法:本实验通过切片法结合静电纺丝及3D打印等技术制备具有取向性的丝素(silk fibroinSF/石墨烯(GrapheneGr)复合材料。显微镜观察材料表面结构形貌。电化学分析提供复合材料的电活性表征。细胞共培养观察该复合材料对神经细胞的影响。结果:不同方法制备的不同取向性复合材料具有不同的导电性,其中3D打印生成取向性的静电纺丝复合材料具有良好的导电性,并且通过体外细胞实验表明其支持培养的施万细胞的存活和生长,揭示其具有良好的生物相容性。结论:结果均表明3D打印取向性复合材料结合了优秀的导电性能与良好的生物相容性等性质,很好地模拟了神经发育的天然神经细胞微环境。

[关键词] 取向性;丝素;石墨烯;神经再生

[中图分类号] Q819 [文献标志码] A

Preparation of oriented bioconductive materials and their effects on nerve regeneration

GONG Jiahuan1, WEI Ziwei2, SHI Jiaqi2, NIU Changmei2, ZHAO Yahong1, YANG Yumin1

(1Jiangsu Key Laboratory of Neuroregeneration; 2School of Medical, Nantong University , Jiangsu 226001)

[Abstract] Objective:The composites with different properties were fabricaed by different methods. The relationship between material structures and nerve regeneration were studied, and the design of this ideal nerve regeneration implant device was expected to provide a solid theoretical basis and a more effective method for the repair of peripheral nerve defects. Method: In this experiment, different silk fibroin (SF) and grapheme(Gr) composites with orientation were prepared by slicing combined with electrospinning, 3D printing. The material surface morphology structures were observed with the microscopes and the electrochemical analysis provided the electrical activity of the composites. The effect of the composites on Schwann cells was observed by the optical microscope. Results: The results showed that different oriented composites have different conductivity. Additionally, the composites had good biocompatibility, and supported the survival and growth of cultured Schwann cells by cell experiments in vitro. Conclusion: In general, all the results indicated that the composites formed by 3D printing had good biocompatibility and excellent electrical conductivity, and simulated the neural microenvironment for nerve regeneration.

[Key words] oriented; silk fibroin; grapheme; nerve regeneration

周围神经损伤后肌萎缩性脊髓侧索硬化症信号通路的变化*

王思嘉**,吴 宸,张 铭,黄 灵,蔡洪玉,李贵才,易

(南通大学神经再生重点实验室,神经再生协同创新中心,江苏226001

[ ] 目的:探讨肌萎缩性脊髓侧索硬化症信号通路在周围神经损伤后的变化。方法:将30SD大鼠随机分为4个实验组和1个对照组,实验组制作坐骨神经夹伤模型,并于伤后014714天取材。使用深度测序结合生物信息学的方法分析坐骨神经组织损伤后肌萎缩性脊髓侧索硬化症信号通路的变化。结果:肌萎缩性脊髓侧索硬化症信号通路在大鼠坐骨神经损伤后显著性趋势随损伤时间而上调,其关键基因的表达量在坐骨神经损伤后不同时间点呈现显著性差异。结论:肌萎缩性脊髓侧索硬化症信号通路参与周围神经的损伤和再生,提示神经损伤再生与神经退行性疾病密切相关。

[关键词] 周围神经损伤;转录组测序;生物信息学;肌萎缩性脊髓侧索硬化症;信号通路

[中图分类号] Q189 [文献标志码] A

High throughput sequencing identifies the involvement of Amyotrophic Lateral Sclerosis signaling in peripheral nerve injury

WANG Sijia, WU Chen, ZHANG Ming, HUANG Ling, CAI Hongyu, LI Guicai, YI Sheng

(Jiangsu Key laboratory of neuroregeneration, Co-innovation Center of Neuroregeneration,

Nantong University , Jiangsu 226001)

[Abstract] Objective: To investigate the changes of Amyotrophic Lateral Sclerosis (ALS) signaling pathway in injured peripheral nerves. Methods: 30 SD rats were randomly divided into four experimental groups and the control group. The sciatic nerves of SD rats were crushed and then collected at 0, 1, 4, 7, and 14 days post injury. Deep sequencing and bioinformatic analysis were combined to analyze ALS signaling pathway. Results: ALS signaling pathway was involved in peripheral nerve injury and the significance of the ALS signaling pathway was elevated with the passage of time. Many critical genes in the ALS signaling pathway were also differentially expressed at different time points after peripheral nerve injury. Conclusion: ALS signaling pathway might be involved in peripheral nerve injury and regeneration, suggesting that peripheral nerve regeneration might be related with neurodegenerative diseases.

[Key words] peripheral nerve injury; transcriptome sequencing; bioinformatics; Amyotrophic Lateral Sclerosis; signaling

狼疮性肾炎患者尿液足细胞和podocalyxin变化及其意义*

刘沙沙1**,宋佳敏2,朱 2,达展云2

1济南市第四人民医院风湿免疫科,山东2500002南通大学附属医院风湿免疫科)

[ ] 目的:探讨狼疮性肾炎(LN)患者足细胞损伤及尿液中podocalyxinPCX)浓度变化与肾脏损害的关系。方法:选择SLE患者60例,其中LN 43例,无肾脏损害SLE 17例,肾病综合征患者(NS20例,健康对照组10例。肾病组63例包括狼疮性肾炎和肾病综合征;非肾病组27例包括系统性红斑狼疮无肾脏受累和健康对照组。根据SLEDAI评分,将LN患者分为中/重度活动组(≥10分)23例和轻度活动/稳定组(<10分)20例;采用ELISA法检测尿液PCX浓度,免疫荧光法检测尿液中足细胞数目;收集LN患者的相关临床和实验室资料,分析各组患者尿液PCX水平及与临床和实验室指标之间的相关性。结果:(1)肾病组患者尿液PCX浓度183.06±79.59 ng/L高于非肾病组16.75±4.97 ng/L;中/重度活动组LN患者尿液PCX浓度270.8±25.50 ng/L高于轻度活动/稳定组90.10±14.08 ng/L和对照组12.74±2.28 ng/L;轻度活动/稳定组尿液PCX浓度高于对照组。(2LNNS患者尿液PCX浓度与24小时尿蛋白定量呈正相关(P<0.01)。(3LN患者尿液PCX浓度与抗ds-DNA抗体水平、SLEDAI评分呈正相关,与C3C4呈负相关(P<0.01)。(4)肾病组尿液足细胞数目及阳性率均高于非肾病组。结论:PCX是足细胞重要的靶蛋白,可作为LN肾损伤和疾病活动度的标志之一。

[关键词] 足细胞;足细胞标记蛋白;系统性红斑狼疮;狼疮性肾炎

[中图分类号] R593.24+1 [文献标志码] A

The change and clinical significance of urine podocyte and podocalyxin protein

in patients with lupus nephritis

LIU Shasha1, SONG Jiamin2, ZHU Jin2, DA Zhanyun2

(1Department of Rheumatology & Immunology, the Forth Hospital of Jinan , Shandong 250000;

2Department of Rheumatology & Immunology, the Affiliated Hospital of Nantong University )

[Abstract] Objective:To explore the relationship between urinary Podocalyxin (PCX) concentration and kidney damage, especially in patients with lupus nephritis (LN) as an assessment of severity and activity, which may be found early renal lesion, to guide further treatment. Methods:The two group included nephropathy group of 63 cases and 27 without nephropathy. The nephropathy group includes lupus nephritis and nephritic syndrome; The non nephropathy group includes systemic lupus erythematosus without renal involvement and healthy control group. 60 cases in SLE patients included 43 cases of LN patients and 17 SLE patients without damage. According to the SLE disease activity index score(SLEDAI) and renal involvement,43 LN patients were divided into different groups:23 patients at high active phase /moderate active phase and 20 at low active phase/stable stage; Patients with nephrotic syndrome (NS) includes 20 cases, and 10 healthy volunteers. By using enzyme-linked immunosorbent assay(ELISA), number concentration and immunofluorescent detection of urinary podocytes and the expression of PCX in urine were detected, and collect the relevant clinical and laboratory data of LN patients were detected. And correlation between PCX levels and clinical symptoms and laboratory indexe was analyzed. Results:(1)The levels of urine PCX in nephropathy group(183.06±79.59 ng/L) were significantly increased compared with the non nephropathy group16.75±4.97 ng/L;The levels of urine PCX were significantly increased in LN patients at high active phase /moderate active phase270.8±25.50 ng/Lcompared with patients at low active phase/stable stage90.10±14.08 ng/Land normal controls12.74±2.28 ng/L; And the low activity/stable group of urine PCX was higher than that of the control group.2LN patients and NS patients with urinary PCX concentrations associated with the 24 hour urinary protein quantitative positiveP<0.01.(3)Urine PCX levels positively correlated with LN disease activity index(SLEDAI) scores,anti ds-DNA antibody levels, and showed a negative correlation with complement C3 and C4P<0.01.(4)The number of podocyte in the nephropathy group is higher than the non nephropathy group. Conclusion:PCX as the foot cell surface marker protein is important, as a kind of important target proteins for the future, how to protect the renal function, delay the progression of disease and treatment related renal diseases including LN provides a new treatment idea.

[Key words] Podocyte; Podocalyxin; Systemic lupus erythematosus; Lupus nephritis