A型肉毒毒素影响透明质酸钠凝胶酶解反应的实验研究  

Experimental Study on the Inhibition of Hyaluronic Acid Gel Hydrolysis by Botulinum Toxin Type A

作　　者：张珍珍 张向东 赵琳[1] 郭倩倩 张选奋[1] ZHANG Zhen-zhen;ZHANG Xiang-dong;ZHAO Lin;GUO Qian-qian;ZHANG Xuan-fen(Department of Plastic Surgery,Second Hospital of Lanzhou University,Lanzhou 730030,Gansu,China)

机构地区：[1]兰州大学第二医院整形外科,甘肃兰州730030

出　　处：《中国美容医学》2020年第5期110-116,152,共8页Chinese Journal of Aesthetic Medicine

摘　　要：目的:探讨A型肉毒毒素(Botulinum toxin type A,BTX-A)对透明质酸钠凝胶(Hyaluronic acid,HA)酶解反应的影响。方法:①分别取海薇和润百颜HA凝胶各5份,每份0.05ml,经2%甲苯胺蓝染色,实验组分别加入2.5U、5.0U、10.0U、20.0U BTX-A,对照组加入生理盐水1.0ml(0U BTX-A);再加入300.0U透明质酸酶,显微镜下观察HA凝胶的酶解状态并采集图片,采用Image J软件计算平均光密度值(OD)及其代谢率;②大鼠背部两侧皮下各注射润百颜HA凝胶0.25ml,左侧真皮层注射BTX-A 5.0U,右侧注射生理盐水0.1ml,分别在注射前(0d)、注射后3、7、14、30、60、90d对注射区域皮肤组织取材,检测透明质酸酶-1(HYAL-1)和透明质酸酶-2(HYAL-2)的表达。结果:①在0U、2.5U、5.0U、10.0U BTX-A作用下,随着BTX-A剂量的增加,HA凝胶的代谢率逐渐减小,润百颜分别在60min、70min、80min、100min达到酶解平衡状态,海薇分别在60min、70min、90min、100min达到酶解平衡状态;20.0U BTX-A对HA凝胶的抑制作用与10.0U BTX-A相似,两种HA凝胶均在100min达到酶解平衡状态。相同酶解条件下,在40min后,润百颜较海薇产品代谢速度增快(P<0.001);②注射后3、7d,BTX-A组大鼠皮肤组织中HYAL-1的表达较生理盐水组降低(P<0.05);注射后3、7、14d,BTX-A组HYAL-2的表达较生理盐水组降低(P<0.05)。结论:BTX-A对透明质酸酶的活性具有抑制作用,并可降低大鼠皮肤组织中HYAL-1、HYAL-2的表达。Objective To investigate the effect of botulinum toxin type A(BTX-A),on the enzymatic degradation of hyaluronic acid gel(HA).Methods①Divided two samples of Matrifi ll and Biohyalux into 5 parts each with a volume of 0.05ml,and stained with 2%toluidine blue solution.The experimental group was respectively added 2.5U,5.0U,10.0U,20.0U BTX-A,and the control group(0U BTX-A)was added with 1.0ml of normal saline.Then added hyaluronidase at a dose of 300.0U,observed the status of HA under the microscope and collect pictures.The average optical density value(OD),and metabolic rate were calculated by Image J software.②Biohyalux was injected into the subcutaneous tissue of the back of rats with a volume of 0.25ml on each side.BTX-A 5.0U was injected into the dermal tissue layer on the left side,and 0.1ml of normal saline was injected on the right side.The skin tissues of the injection area were taken before the injection(0d),and 3,7,14,30,60,90 days after the injection for detected the expression of HYAL-1,HYAL-2.Results①Under the effects of BTX-A at 0U,2.5U,5.0U and 10.0U,as the dose of BTX-A increased,the metabolic rate of HA gradually decreased.The product of Biohyalux basically reached an equilibrium state of enzyme degradation at 60min,70min,80min and 100min.And Matrifi ll basically reached an equilibrium state of enzyme degradation at 60min,70min,90min and 100min.The inhibitory effect of 20.0U BTX-A on HA gel was similar to that of 10.0U BTX-A.Both HA gels reached the enzymatic hydrolysis equilibrium state at 100min.Under the same enzymolysis conditions,Biohyalux had a faster metabolism rate than Matrifi ll after 40min(P<0.001).②On the 3 and 7 days after injection,the expression of HYAL-1 in the skin tissues of the BTX-A group was lower than those of the normal saline group(P<0.05).On the 3,7 and 14 days,the HYAL-2 expression of the BTX-A group was lower than those of the normal saline group(P<0.05).Conclusion BTX-A has an inhibitory effect on the activity of hyaluronidase and reduces the expression of HYAL-1

关 键 词：A型肉毒毒素 透明质酸钠凝胶 透明质酸酶 酶解作用 

分 类 号：Q946[生物学—植物学]