CAJ | 학술논문

背景：许多研究表明，川芎嗪对细胞及异体神经有一定的免疫抑制作用，能减轻炎症细胞的浸润程度，降低异体神经移植体的免疫排斥反应。 　　目的：观察川芎嗪对组织工程化神经桥接物生物学特性的影响。 　　方法：采用化学法萃取Wistar大鼠去细胞坐骨神经，构建种植基因修饰神经干细胞的组织工程化神经桥接物，分别于添加与未添加200 mg/L川芎嗪的完全培养基中培养1周，观察细胞分布与支架结构。取SD大鼠24只，随机均分为4组，分别于背部皮下植入4℃保存1周的Wistar大鼠坐骨神经(异种神经组)、常规完全培养基培养的组织工程化神经(对照组)、含200 mg/L川芎嗪完全培养基培养的组织工程化神经(实验组)及同种异体SD大鼠坐骨神经(同种异体神经组)，7 d后取出各组神经行苏木精-伊红染色光镜观察及CD4+和CD8+细胞浸润数量检测。 　　结果与结论：去细胞神经基本无细胞成分，神经外膜和细胞基底膜结构均较完整；注射神经干细胞后，可见神经外膜下和束膜间有较多细胞成分，分布不均；培养1周后，两组培养基均有较多增殖与分化的神经细胞，以添加川芎嗪的完全培养基组效果更明显。皮下包埋后光镜下淋巴细胞浸润由多到少的顺序为：异种神经组、对照组、实验组、同种异体神经组。表明川芎嗪可降低组织工程化神经的免疫原性，但不破坏组织工程神经的三维结构。
배경：허다연구표명，천궁진대세포급이체신경유일정적면역억제작용，능감경염증세포적침윤정도，강저이체신경이식체적면역배척반응。 　　목적：관찰천궁진대조직공정화신경교접물생물학특성적영향。 　　방법：채용화학법췌취Wistar대서거세포좌골신경，구건충식기인수식신경간세포적조직공정화신경교접물，분별우첨가여미첨가200 mg/L천궁진적완전배양기중배양1주，관찰세포분포여지가결구。취SD대서24지，수궤균분위4조，분별우배부피하식입4℃보존1주적Wistar대서좌골신경(이충신경조)、상규완전배양기배양적조직공정화신경(대조조)、함200 mg/L천궁진완전배양기배양적조직공정화신경(실험조)급동충이체SD대서좌골신경(동충이체신경조)，7 d후취출각조신경행소목정-이홍염색광경관찰급CD4+화CD8+세포침윤수량검측。 　　결과여결론：거세포신경기본무세포성분，신경외막화세포기저막결구균교완정；주사신경간세포후，가견신경외막하화속막간유교다세포성분，분포불균；배양1주후，량조배양기균유교다증식여분화적신경세포，이첨가천궁진적완전배양기조효과경명현。피하포매후광경하림파세포침윤유다도소적순서위：이충신경조、대조조、실험조、동충이체신경조。표명천궁진가강저조직공정화신경적면역원성，단불파배조직공정신경적삼유결구。
BACKGROUND:Increasing studies have shown that tetramethylpyrazine can play certain immunosuppressive effects on cels and alogeneic nerves, reduce the infiltration of inflammatory cels, and reduce the body’s immune rejection to nerve alografts. OBJECTIVE:To observe the effect of tetramethylpyrazine on the biological characteristics of tissue-engineered nerve bridging materials. METHODS:Acelular sciatic nerves were extracted from Wistar rats using chemical method to construct gene-modified nerve alografts that were cultured in complete medium containing 200 mg/L tetramethylpyrazine or not for 1 week to observe the cel distribution and structure. Twenty-four healthy adult male Sprague-Dawley rats were enroled and randomly divided into four groups: in nerve xenograft group, sciatic nerve segments from Wistar rats cultured at 4℃ for 1 week were implanted subcutaneously into the back of Sprague-Dawley rats; in control group, tissue-engineered nerve segments cultured in the complete medium were implanted; in experimental group, tissue-engineered nerve segments cultured in the complete medium containing 200 mg/L tetramethylpyrazine were implanted; in nerve alograft group, nerve alografts from Sprague-Dawley rats were implanted. After 7 days, nerve segments were taken out for hematoxylin-eosin staining under light microscope, and the number of CD4+ and CD8+ lymphocytes was counted. RESULTS AND CONCLUSION:The acelular nerve segments with no celular constituents had intact epineurium and basement membrane. After injection of nerve cels, there were more celular components that distributed unevenly beneath the epineurium and between the perineurium. After 1 week of culture, more nerve cels proliferated and differentiated in the complete medium containing 200 mg/L tetramethylpyrazine or not, especialy in the complete medium containing 200 mg/L tetramethylpyrazine. Under the light microscope, the degree of lymphocyte infiltration ranged from more to less: nerve xenograft group, control group, experimental group, nerve alograft group. These findings indicate that tetramethylpyrazine can reduce tissue-engineered nerve immunogenicity, but cannot destroy the three-dimensional structure of tissue-engineered nerves.