CAJ | 학술논문

目的探索透明质酸-半乳糖化壳聚糖复合支架(HGC)联合牛磺酸(Tau)对脑创伤(TBI)大鼠的治疗作用.方法选择成年SD大鼠40只,随机分为假手术组(Sham组)、脑创伤组(TBI组)、透明质酸-半乳糖化壳聚糖复合支架组(HGC组)、透明质酸-半乳糖化壳聚糖复合支架与牛磺酸联合治疗组(HGC-Tau组),每组10只.TBI、HGC及HGC-Tau组采用液压打击法在大鼠左侧大脑半球制作TBI模型,Sham组仅在相同位置开骨窗,不予打击.HGC组于造模后7d将30μl HGC植入大鼠脑皮质损伤区,HGC-Tau组以同样方法植入相同体积HGC与牛磺酸的混合物.TBI后1、3、7、10、14、21 d进行改良型神经功能评分(mNSS),其中17～21 d进行Morris水迷宫(MWM)实验.TBI后28 d采用实时定量PCR及免疫荧光染色检测胶质纤维酸性蛋白质(GFAP)的基因及蛋白表达水平.结果 HGC-Tau组mNSS评分在TBI后14d及21d显著低于TBI组(P＜0.05),HGC组各时间点评分与TBI组相比,差异均无统计学意义(P＞0.05),HGC-Tau组与HGC组相比,评分在TBI后14、21 d显著降低(P＜0.05);HGC-Tau组MWM检测目标象限百分比从创伤后18d开始明显大于TBI组(P＜0.05),HGC组创伤后21 d目标象限百分比显著大于TBI组(P＜0.05),HGC-Tau组仅在18、19 d目标象限百分比明显大于HGC组(P＜0.05);HGC组及HGC-Tau组TBI后28 dGFAP mRNA及蛋白表达量与TBI组相比显著下降(P＜0.05),且较HGC组,HGC-Tau组下降更加明显(P＜0.05).结论单纯使用HGC仅能抑制创伤性脑损伤后胶质细胞增生,对神经功能改善无明显作用.HGC联合Tau治疗可抑制重型脑损伤后胶质细胞增生,改善神经功能,表明生物材料与药物联合治疗脑外伤具有一定的应用前景.
목적 탐색투명질산-반유당화각취당복합지가(HGC)연합우광산(Tau)대뇌창상(TBI)대서적치료작용.방법 선택성년SD대서40지,수궤분위가수술조(Sham조)、뇌창상조(TBI조)、투명질산-반유당화각취당복합지가조(HGC조)、투명질산-반유당화각취당복합지가여우광산연합치료조(HGC-Tau조),매조10지.TBI、HGC급HGC-Tau조채용액압타격법재대서좌측대뇌반구제작TBI모형,Sham조부재상동위치개골창,불여타격.HGC조우조모후7d장30μl HGC식입대서뇌피질손상구,HGC-Tau조이동양방법식입상동체적HGC여우광산적혼합물.TBI후1、3、7、10、14、21 d진행개량형신경공능평분(mNSS),기중17～21 d진행Morris수미궁(MWM)실험.TBI후28 d채용실시정량PCR급면역형광염색검측효질섬유산성단백질(GFAP)적기인급단백표체수평.결과 HGC-Tau조mNSS평분재TBI후14d급21d현저저우TBI조(P＜0.05),HGC조각시간점평분여TBI조상비,차이균무통계학의의(P＞0.05),HGC-Tau조여HGC조상비,평분재TBI후14、21 d현저강저(P＜0.05);HGC-Tau조MWM검측목표상한백분비종창상후18d개시명현대우TBI조(P＜0.05),HGC조창상후21 d목표상한백분비현저대우TBI조(P＜0.05),HGC-Tau조부재18、19 d목표상한백분비명현대우HGC조(P＜0.05);HGC조급HGC-Tau조TBI후28 dGFAP mRNA급단백표체량여TBI조상비현저하강(P＜0.05),차교HGC조,HGC-Tau조하강경가명현(P＜0.05).결론 단순사용HGC부능억제창상성뇌손상후효질세포증생,대신경공능개선무명현작용.HGC연합Tau치료가억제중형뇌손상후효질세포증생,개선신경공능,표명생물재료여약물연합치료뇌외상구유일정적응용전경.
Objective To investigate the treatment effect of hyaluronic acid-galactosylated chitosan scaffold (HGC) combined with taurine (Tau) in traumatic brain injury (TBI) after implanted to the injured cortex in rat.Methods Forty SD rats were equally divided into Sham group,TBI group,HGC treated group (HGC group) and HGC plus taurine treated group (HGC-Tau group).Rats in TBI,HGC and HGC-Tau group were subjected to severe traumatic injury of left brain by lateral fluid percussion device,while sham control rats underwent the same surgical procedure weren't exposed to percussion injury.Rats in HGC group were implanted with 30 μl HGC at brain cortical lesion site 7 d after TBI immediately.HGC-Tau group rats were implanted with HGC and taurine with same volume at the same site post-TBI at the same time.Modified neurological severity score (mNSS) test was conducted at 1,3,7,10,14 and 21 d after TBI.Morris water maze (MWM) was used to detect the spatial reference memory for 5 consecutive days starting on 17 d after TBI.RT-PCR and immunohistochemistry for glial fibrillary acidic protein (GFAP) were performed at 28 d after TBI.Results The HGC-Tau treated group had significantly improved functional outcome at 14 and 21 d after TBI measured by mNSS test,compared with both TBI group and HGC group (P＜0.05),While there was no significant difference between HGC group and TBI group in mNSS test at all time points (P＞0.05).HGC-Tau treatment could improve spatial learning ability of rats at all time points except 17 d after TBI in MWM test (P＜0.05).The mean ratios of time spent in the target quadrant were significantly higher in the HGC-Tau treated group when compared to the TBI control group at 21 d (P＜0.05).The mean percentages of rats in HGC-Tau treated group were significantly higher than that of the HGC group at 18 and 19 d after TBI (P＜0.05).RTPCR showed that the rats in both HGC-Tau group and HGC group have lower expression of GFAP mRNA compared to the rats in TBI group (P＜0.05).GFAP mRNA expression in HGC-Tau group was higher than that in HGC group (P＜0.05).Immuocytochemical fluorescent staining showed the same tendency.Conclusions Pure HGC can only inhibit reactive astrocytes post-TBI,while HGC combined with taurine can inhibit reactive astrocytes and promote the recovery of neural function in rats with severe TBI.Biomaterials combined with drug have a certain application prospect.