摘要
目的:探究右归饮联合经转化生长因子-β(TGF-β1)转染骨髓间充质干细胞用于修复大鼠软骨组织的效果。方法:选取40只SD大鼠,采用手术钻孔的方法将大鼠制成软骨组织损伤模型;采用随机数字表法分为4组,每组10只,其中A组为空白组,仅仅制成软骨组织损伤模型不作任何处理;B组为单纯右归饮组采用右归饮灌胃;C组为单纯复合体组,在缺损中植入干细胞3D生物支架;D组为右归饮结合间充质干细胞(BMSCs)3D生物支架实验组,关节软骨缺损中嵌入干细胞3D生物支架同时每天中药右归饮灌胃,持续8周。术后4、8周行空气栓塞分批处死,采用HE染色分别观察各组大鼠软骨组织形态学变化;采用病理学Mankin评分比较各组大鼠软骨组织情况;采用生物力学测试机测定各组大鼠软骨组织杨氏弹性模量;采用实时聚合酶链反应(RT-PCR)和蛋白质印迹(WB)方法测定软骨细胞中Ⅱ型胶原及可聚蛋白多糖的mRNA及蛋白表达。结果:HE染色结果显示,相比A组,B组和C组大鼠软骨组织炎性浸润及坏死显著改善,相比B组和C组,D组大鼠基本无炎性浸润,大鼠软骨组织已基本修复。各组内大鼠相比4周时,8周后大鼠Mankin评分均显著降低(P<0.05);相比A组,B组和C组大鼠Mankin评分显著降低,载荷-位移、应力-应变、弹性模量、Ⅱ型胶原及可聚蛋白多糖的mRNA及蛋白相对表达水平均显著升高(P<0.05);相比,B组和C组,D组Mankin评分显著降低,载荷-位移、应力-应变、弹性模量、Ⅱ型胶原及可聚蛋白多糖的mRNA及蛋白相对表达水平均显著升高(P<0.05)。结论:右归饮联合经TGF-β1转染BMSCs可促进大鼠膝关节损伤后的修复,可能与促进Ⅱ型胶原及可聚蛋白多糖表达有关。
Objective:To explore the effect of using Yougui Decoction combined with transforming growth factor-β(TGF-β1)transfected bone mesenchymal stem cells(BMSCs)in repairing cartilage tissues for rats.Methods:Forty SD rats were enrolled and made into cartilage tissue injury models by surgical drilling.They were divided into group A,B,C and D by the random number table method,10 cases in each group.In the group A(the blank group),they were only made into cartilage tissue injury models and not treated.In the group B(the Yougui Decoction group),they were given intragastric administration of Yougui Decoction.In the group C(the complex group),stem cell 3 D bioscaffold was implanted at the defect sites.In the group D(the Yougui Decoction combined with BMSCs 3 D bioscaffold experiment group),stem cell 3 D bioscaffold was embedded at articular cartilage defects sites,and intragastric administration of Yougui Decoction was given for 8 weeks.At 4 and 8 weeks after surgery,air embolization was performed to kill them in batches.The morphological changes of cartilage tissues in each group were observed by HE staining.The cartilage tissues situations were compared among all groups by pathological Mankin scores.The Ylastic Young’s modulus of cartilage tissues in each group were measured by biomechanics test machine.The mRNA and protein expression of type Ⅱ collagen and polymerizable proteoglycan in chondrocytes were measured by real-time polymerase chain reaction(RT-PCR)and Western blot(WB).Results:HE staining results showed that compared with the group A,inflammation infiltration and necrosis of cartilage tissues were significantly improved in the group B and C.Compared with the group B and C,there was almost no inflammation infiltration in the group D,and cartilage tissues were basically repaired.Compared with those at 4 weeks after surgery,the Mankin scores were significantly decreased at 8 weeks after surgery in each group(P<0.05).Compared with the group A,Mankin scores were significantly decreased in the group B and C,while load-displacement,stress-strain,elastic modulus,relative expression levels of type Ⅱ collagen and polymerizable proteoglycan mRNA and protein were significantly increased(P<0.05).Compared with the group B and C,the Mankin scores were significantly decreased in the group D,while load-displacement,stress-strain,elastic modulus,relative expression levels of type II collagen and polymerizable proteoglycan mRNA and protein were significantly increased(P<0.05).Conclusion:Yougui Decoction combined with TGF-β1 transfected BMSCs can promote the repair of knee injury in rats,which may be related to promoting the expressions of type II collagen and aggrecan.
作者
罗栋新
黄英
刘承俊
LUO Dongxin;HUANG Ying;LIU Chengjun(Department of Orthopedics and Traumatology,Tieling Hospital of Traditional Chinese Medicine,Tieling Liaoning112000,China)
出处
《四川中医》
2022年第1期49-53,共5页
Journal of Sichuan of Traditional Chinese Medicine
基金
辽宁省自然科学基金(编号:2013021020)
关键词
软骨组织
右归饮
骨髓间充质干细胞
转化生长因子-Β
Cartilage tissue
Yougui Decoction
Bone mesenchymal stem cell
Transforming growth factor-β
