镉致鸡胚胫骨软骨发育不良的研究  

Effect of cadmium on tibial dyschondroplasia in chicken embryo

作　　者：王果帅 刘丹 沈小兰 张雪晴 李雅文 袁燕[1,3,4] 邹辉 卞建春[1,3,4] 刘学忠 刘宗平 顾建红[1,3,4] WANG Guoshuai;LIU Dan;SHEN Xiaolan;ZHANG Xueqing;LI Yawen;YUAN Yan;ZOU Hui;BIAN Jianchun;LIU Xuezhong;LIU Zongping;GU Jianhong(College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China;Hechen Animal Husbandry and Veterinary Station,Xinghua City,Taizhou 225733,China;Jiangsu Province Key Laboratory of Zoonosis,Yangzhou 225009,China;Jiangsu Universities Collaborative Innovation Center for Prevention and Control of Important Animal Diseases and Zoonosis,Yangzhou 225009,China)

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]江苏省兴化市合陈畜牧兽医站,江苏泰州225733 [3]江苏省人兽共患病学重点实验室,江苏扬州225009 [4]江苏高校动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《畜牧与兽医》2022年第1期112-117,共6页Animal Husbandry & Veterinary Medicine

基　　金：江苏省自然科学基金(BK20181452);国家自然科学基金(31872534,31872533);江苏高校优势学科建设工程项目(PAPD)。

摘　　要：为了研究镉对鸡胚胫骨软骨发育的影响,选取体重相近的SPF鸡胚120枚随机分为4组,每组30枚,孵化6 d后在鸡胚气室打孔,其中对照组向卵黄囊注射PBS 50μL,其他3组分别向卵黄囊注射0.05、0.1和0.2 mg/mL醋酸镉50μL,处理10 d,通过石蜡切片HE染色、阿利新蓝染色观察胫骨软骨发育情况;扫描电镜观察软骨表面结构;荧光定量PCR检测软骨细胞标志性基因ACAN、COL2A1、SOX9、COL10A1、MMP-13的表达。结果显示:0.05 mg/mL镉组胫骨软骨细胞分泌的蛋白多糖增多且细胞形态正常,而≥0.1 mg/mL镉组细胞分泌的蛋白多糖减少、细胞核固缩溶解,镉处理组软骨表面胶原纤维变得疏松,排序紊乱。0.05 mg/mL镉组与对照组相比,软骨分化前期相关因子ACAN、SOX9、COL2A1显著升高(P<0.05),但软骨分化后期相关转录因子MMP-13、COL10A1表达水平显著(P<0.05)或极显著(P<0.01)降低,≥0.1 mg/mL镉组均呈抑制作用且呈剂量依赖性。结果表明,低浓度镉(0.05 mg/mL)能促进软骨早期分化,但高浓度镉(≥0.1 mg/mL)会损伤软骨表面结构、破坏软骨细胞生理功能、抑制软骨分化,对软骨生长发育有抑制作用。In order to identify the effect of cadmium on the development of tibial cartilage of chicken embryos,120 SPF embryo samples of chickens with similar body weight were obtained and randomly divided into four groups,30 in each group.After 6 days of incubation,holes were punched in the air chamber of the chicken embryos,in which the control group were injected with 50μL PBS solvent into the yolk sac,and the other three groups with 50μL 0.05,0.1 and 0.2 mg/m L cadmium acetate into the yolk sac,respectively.The test lasted for 10days.The tibia was observed by paraffin section HE staining and allicin blue staining.Scanning electron microscope was used to observe the cartilage surface structure.QRT-PCR was used to detect the expression of the marker genes ACAN,COL2A1,SOX9,COL10A1 and MMP-13 in chondrocytes.The results showed that the proteoglycan secreted by tibial chondrocytes in the 0.05 mg/m L cadmium group increased and their cell morphology was normal,while the proteoglycan secreted by cells in the≥0.1 mg/m L cadmium groups decreased,and their nuclei were pyknosis and dissolution,and the collagen fibers on the cartilage surface in the cadmium treated group became loose and disordered.Compared with the control group,the related factors ACAN,SOX9 and COL2A1 in the 0.05 mg/m L cadmium group were significantly increased (P<0.05),but the expression levels of the related transcription factors MMP-13 and COL10A1 in the late stage of cartilage differentiation were significantly (P<0.05) or extremely significantly (P<0.01) decreased,and all the groups treated with≥0.1 mg/m L cadmium showed inhibitory effect and dosage.These results indicated that low concentration of cadmium (0.05 mg/m L) could promote the early differentiation of cartilage,but high concentration of cadmium (≥0.1 mg/m L) could damage the surface structure of cartilage,destroy the physiological function of chondrocytes and inhibit the differentiation of cartilage.

关 键 词：鸡 镉 胫骨软骨 阿利新蓝染色 分化 

分 类 号：S859.8[农业科学—临床兽医学]