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目的:探索丹参注射液对脂肪源性干细胞(ASCs)成软骨的促进作用。方法:①获取C57小鼠ASCs并培养传代,取第4代用于体外实验。分为空白组、诱导组与丹参注射液组,分别予常规培养液、成软骨诱导液及成软骨诱导液加丹参注射液干预,培养14 d后采用定量PCR检测ASCs体外Ⅱ型胶原(CollagenⅡ)的表达。②取2月龄裸鼠15只,在髌股关节面用克氏针造成软骨缺损。随机分为模型组、诱导细胞复合纤维蛋白胶植入组和丹参注射液干预细胞复合纤维蛋白胶植入组,术后12周切取膝关节缺损处,行HE染色。结果:①通过体外对CollagenⅡmRNA表达的研究证实,诱导组和丹参注射液组均可促进ASCs向软骨细胞分化,尽管两者数值相比没有统计学差异(P>0.05),但与诱导组相比丹参注射液组可以提高CollagenⅡmRNA的表达水平。②体内修复的HE染色表明,丹参注射液组和诱导组与纤维蛋白胶复合后均具有促进软骨再生,其中丹参注射液干预细胞组的再生软骨优于诱导组。结论:丹参注射液具有促进ASCs向软骨细胞分化的功能,与支架材料复合后可以较好地修复软骨缺损,这将为中药应用软骨组织工程的发展奠定基础。
Objective: To explore the promoting effects of Danshen injection on cartilage of adipose derived stem cells (ASCs). Methods: ① C57 mouse ASCs were obtained and cultured for passage, and the 4th generation was used for in vitro experiments. The rats were randomly divided into blank group, induction group and Salvia miltiorrhiza injection group. The cells were treated with routine culture medium, chondrogenic induction solution and chondrogenic induction solution plus salvia miltiorrhiza injection respectively. After 14 d of culture, collagenase Ⅱ (ASCs) expression. ② Take 2-month-old nude mice 15, in the patellofemoral joint surface caused by Kirschner wire cartilage defects. The rats were randomly divided into model group, induced cell composite fibrin glue implantation group and Danshen injection intervened cell composite fibrin glue implantation group. The knee joint defects were excised at 12 weeks after operation, and HE staining was performed. Results: (1) The study of Collagen II mRNA expression in vitro showed that both induction group and Danshen injection group could promote the differentiation of ASCs to chondrocytes. Although there was no significant difference between the two groups (P> 0.05), compared with the induction group Salvia injection group can increase the expression of Collagen Ⅱ mRNA levels. ② HE staining of in vivo repair showed that Danshen injection group and induced group and fibrin glue had the promotion of cartilage regeneration, of which Danshen injection intervention group cells regenerated cartilage better than the induction group. Conclusion: Salvia miltiorrhiza injection can promote the differentiation of ASCs to chondrocytes, which can repair cartilage defects well after being combined with scaffolds, which will lay the foundation for the development of cartilage tissue engineering by traditional Chinese medicine.