活血化瘀汤对股骨骨折大鼠骨痂组织胶原表达水平及促进愈合机制分析
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摘要
目的:探讨活血化瘀汤对股骨骨折大鼠骨痂组织胶原表达水平及促进愈合机制。方法:选取由安康市人民医院和陕西省人民医院实验动物中心提供清洁级雄性SD大鼠43只,依据随机数字表法将大鼠分成两组,观察组(22只)、对照组(21只),股骨中段使用线锯切断制备股骨骨折模型,造模后第二天观察组活血化瘀汤灌胃,对照组使用同剂量生理盐水灌胃;灌胃四周后使用颈椎脱臼法将大鼠处死,察看股骨骨痂生长和骨折愈合状况,同时行HE染色察看大鼠骨痂组织结构,免疫组化检测大鼠治疗后7天、14天、21及28天骨痂组织内Ⅰ与Ⅱ型胶原表达状况;比色法检测血清磷(P)、钙(Ca)、碱性磷酸酶(ALP)含量,ELISA检测血清生长激素(GH)含量。结果:治疗28天时观察组大鼠骨痂组织内钙化状况和骨小梁结构则显著优于对照组,治疗后7天、14天、21天、28天观察组大鼠骨痂组织内Ⅰ型及Ⅱ型胶原表达水平均高于对照组,差异有统计学意义(P<0. 05);治疗后21天、28天观察组大鼠血清GH、P、Ca及ALP水平均高于对照组,差异有统计学意义(P<0. 05)。结论:骨折前期活血化瘀汤可加速骨痂组织内Ⅰ型与Ⅱ型胶原合成,更有利于纤维-软骨性骨痂产生。
Objective: To analyze the expression of collagen in callus tissue of rats with femoral fracture and the mechanism of promoting healing. Methods: Forty-three male SD rats were divided into two groups according to the random number table,the observation group( n = 22) and the controlled group( n = 21). Preparation of femoral fracture models with cut off,the observation group was given Huoxue Huayu Decoction at the second day with gavage,the controlled group used the same dose of normal saline gavage,four weeks after the use of cervical dislocation method,the rats were sacrificed to observe the femoral callus growth and at the same time,the callus tissue of rat was observed by HE staining. The expression of type I and type II collagen in callus were detected by immunohistochemistry at 7,14,21 and 28 days after treatment. Serum phosphorus( P),calcium( Ca) and alkaline phosphatase( ALP) levels were detected by colorimetric assay. Serum growth hormone( GH) levels were measured by ELISA. Results: At the 28 th day of treatment,calcification and trabecular structure in the callus tissue of the observation group were significantly better than that of the controlled group. At 7,14,21 and 28 days after treatment,the type Ⅰ and type ⅡCollagen expression levels were higher than the controlled group,the difference was statistically significant( P < 0. 05). The levels of serum GH,P,Ca and ALP in the observation group were higher than those in the controlled group 21 and 28 days after treatment( P < 0. 05). Conclusion: Huoxue Huayu Decoction can accelerate the synthesis of type I and type II collagen in callus tissue,and is more conducive to the production of fiber-cartilage callus.
Objective: To analyze the expression of collagen in callus tissue of rats with femoral fracture and the mechanism of promoting healing. Methods: Forty-three male SD rats were divided into two groups according to the random number table,the observation group( n = 22) and the controlled group( n = 21). Preparation of femoral fracture models with cut off,the observation group was given Huoxue Huayu Decoction at the second day with gavage,the controlled group used the same dose of normal saline gavage,four weeks after the use of cervical dislocation method,the rats were sacrificed to observe the femoral callus growth and at the same time,the callus tissue of rat was observed by HE staining. The expression of type I and type II collagen in callus were detected by immunohistochemistry at 7,14,21 and 28 days after treatment. Serum phosphorus( P),calcium( Ca) and alkaline phosphatase( ALP) levels were detected by colorimetric assay. Serum growth hormone( GH) levels were measured by ELISA. Results: At the 28 th day of treatment,calcification and trabecular structure in the callus tissue of the observation group were significantly better than that of the controlled group. At 7,14,21 and 28 days after treatment,the type Ⅰ and type ⅡCollagen expression levels were higher than the controlled group,the difference was statistically significant( P < 0. 05). The levels of serum GH,P,Ca and ALP in the observation group were higher than those in the controlled group 21 and 28 days after treatment( P < 0. 05). Conclusion: Huoxue Huayu Decoction can accelerate the synthesis of type I and type II collagen in callus tissue,and is more conducive to the production of fiber-cartilage callus.
引文
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[16]柏小金,徐林,骆旭东,等.构建股骨骨折合并脑损伤模型大鼠低氧诱导因子1α和核心结合因子α1的表达[J].中国组织工程研究,2015,19(18):2800~2806
[2]王斌,罗毅文,黄永铨,等.补肾活血汤促骨折端间充质干细胞体外迁移及CCR2表达的研究[J].中国中医骨伤科杂志,2015,11(8):1~5
[3] Aydin A,Halici Z,Akpinar E,et al. What is the role ofbosentan in healing of femur fractures in a rat model?[J]. Journal of BoneMineral Metabolism,2015,33(5):496~506
[4]庞学丰,舒建龙,李玉玲,等.寒痹康汤对胶原诱导性关节炎大鼠血清VEGF的影响[J].陕西中医,2015,3(5):616~618
[5]王琳娜,陈常勇,陈小永,等.当归补血汤对尿毒症大鼠心肌纤维化组织瞬时受体电位M7通道表达及胶原蛋白合成的影响[J].中国现代医学杂志,2015,25(30):7~12
[6]周缜,蓝青,李洪波,等.壮筋续骨汤对骨折大鼠血清内源性生长因子水平的影响及意义[J].中国中西医结合外科杂志,2015,21(5):479~484
[7] Yang L,Rui Y,Cheng T Y,et al. Effects of Sclerostin Anti-body on the Healing of Femoral Fractures in Ovariectomised Rats[J]. Cal-cified Tissue International,2016,98(3):263~274
[8]王希强,孙仁光,孙仕润,等.丹参接骨胶囊对闭合性股骨骨折模型大鼠骨折愈合的影响[J].中国组织工程研究,2016,20(5):622~627
[9] Guimar? es A,Butezloff M M,Zamarioli A,et al. Nandrolonedecanoate appears to increase bone callus formation in young adult rats aftera complete femoral fracture[J]. Acta Cirurgica Brasileira, 2017, 32(11):924~928
[10]童凯,尹宏,卞勇.活血散干预大鼠骨折模型早期骨愈合的实验研究[J].贵州医药,2017,41(4):355~357
[11]龙朝阳,李凤珍,梁江洪,等.清毒伸筋汤对胶原诱导性关节炎大鼠血清TNF-α、IL-1β水平的影响[J].中医学报,2017,32(1):93~95
[12]宁驰,周缜,侯海青,等.壮筋续骨汤对骨折大鼠骨痂组织中CTR和CBFα1表达的影响[J].中国中医骨伤科杂志,2016,9(7):1~6
[13]汪雷,刘林娟,肖黎,等.脊髓损伤合并股骨骨折大鼠模型中leptin的表达[J].实用医学杂志,2015,31(22):3688~3690
[14]肖增兵,陈通,付爱军,等.构建颅脑损伤合并股骨闭合性骨折大鼠模型:骨痂中降钙素基因相关肽和胰岛素样生长因子1的表达[J].中国组织工程研究,2015,19(5):657~661
[15] Saita Y,Ishijima M,Mogami A,et al. The incidence of andrisk factors for developing atypical femoral fractures in Japan[J]. Journalof Bone Mineral Metabolism,2015,33(3):311~318
[16]柏小金,徐林,骆旭东,等.构建股骨骨折合并脑损伤模型大鼠低氧诱导因子1α和核心结合因子α1的表达[J].中国组织工程研究,2015,19(18):2800~2806