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作 者:朱彩玉 周正新[2] 顾一帆 韩士鼎 徐寰 王壮壮 朱磊[2] ZHU Cai‐yu;ZHOU Zheng‐xin;GU Yi‐fan;HAN Shi‐ding;XU Huan;WANG Zhuang‐zhuang;ZHU Lei(The First Clinical Medical College of Anhui University of Chinese Medicine,Hefei 230031,China;The First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031,China)
机构地区:[1]安徽中医药大学第一临床医学院,安徽合肥230031 [2]安徽中医药大学第一附属医院,安徽合肥230031
出 处:《海南医学院学报》2022年第19期1451-1457,共7页Journal of Hainan Medical University
基 金:安徽省教育厅高校自然科学研究重点项目(KJ2020A0403);安徽省自然科学基金面上项目(2008085MH281)。
摘 要:目的:探讨骨痹通消颗粒对糖皮质激素致股骨头坏死过程中血管形成及细胞凋亡的干预作用。方法:将30只实验用新西兰兔随机分组,除对照组外,使用脂多糖联合糖皮质激素建立模拟激素性股骨头坏死动物模型。造模成功后,实验组给予骨痹通消颗粒灌胃治疗,模型组和对照组则给予等量生理盐水灌胃,1次/d,持续8周后处死实验动物,取出各组兔双侧股骨头进行苏木精-伊红染色并对蛋白CD34、CYR61及VEGF进行免疫组化染色定位;采用TUNEL染色法观察股骨头区细胞凋亡情况;WB检测凋亡蛋白Bax及Bcl-2表达水平;RT-qPCR分析VEGF、eNOs、Bax及Bcl-2等基因表达情况。结果:与模型组相比,骨痹通消颗粒能够减少骨组织中空骨陷窝数和凋亡阳性细胞数(P<0.01),上调股骨头组织中CD34、CYR61及VEGF表达。与对照组相比,模型组中促凋亡蛋白Bax的表达量增多且抗凋亡蛋白Bcl-2、VEGF及eNOs mRNA的表达量下降明显(P<0.01),实验组经骨痹通消颗粒干预治疗后得到相反的结果(P<0.05)。结论:骨痹通消颗粒能够提高激素性股骨头坏死兔模型骨组织中成血管基因表达量并调控凋亡相关蛋白,促进其成血管分化并抑制细胞凋亡,进而发挥治疗激素性股骨头坏死的作用。Objective:To investigate the effect of Gubi Tongxiao granules on vascular formation and cell apoptosis in the process of glucocorticoid-induced femoral head necrosis.Methods:Thirty experimental New Zealand rabbits were randomly divided into groups. Except the control group,the animal model of steroid-induced femoral head necrosis was established by lipopolysaccharide combined with glucocorticoids. After successful modeling,the experimental group was given Gubi Tongxiao granules intragastric treatment,the model and control group were given the same amount of normal saline intragastric treatment,once a day,for 8 weeks,the experimental animals were sacrificed,the bilateral femoral head of each group was taken out for hematoxylin-eosin staining,and protein CD34,CYR61 and VEGF were immunohistochemical staining localization. TUNEL staining was used to observe the apoptosis of femoral head cells. The expression levels of apoptosis proteins Bax and Bcl-2 were detected by WB. The expression of VEGF eNOs Bax and Bcl-2 genes was analyzed by RT-qPCR.Results:Gubi Tongxiao granules reduced the number of hollow bone lacunae and apoptotic positive cells(P<0.01),and up-regulated the expressions of CD34,CYR61 and VEGF in femoral head tissue. Compared with control group,the expression of pro-apoptotic protein Bax increased and the expression of anti-apoptotic protein Bcl-2,VEGF and eNOs mRAN decreased significantly in the model group(P<0.01),the opposite results were obtained in the experimental group after Gubi Tongxiao granule intervention(P<0.05).Conclusion:Gubi Tongxiao granules can improve the expression of angiogenic genes and regulate apoptosis-related proteins in the bone tissue of rabbit model with steroid-induced necrosis of femoral head,promote angiogenic differentiation and inhibit apoptosis,and thus achieve the effect of treating steroid-induced necrosis of femoral head.
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