文献类型：期刊文献

中文题名：右归丸对膝骨关节炎模型鼠软骨组织凋亡和基质降解调控基因表达的影响

英文题名：Effects of Youguiwan on gene expression of apoptosis and matrix degradation of chondrocytes in rats with knee osteoarthritis

作者：颜春鲁[1,2,3,4];安方玉[1,2,3,4];汪永锋[1];刘永琦[1,2,4];蔺兴遥[1,4];蒋国凤[1];赵磊[1,3];蒋溪媛[1];吕学颖[1];石彪[1]

第一作者：颜春鲁

机构：[1]甘肃中医药大学,兰州730000;[2]甘肃省高校重大疾病分子医学与中医药防治研究重点实验室,兰州730000;[3]甘肃省高校中(藏)药化学与质量研究省级重点实验室,兰州730000;[4]敦煌医学与转化省部共建教育部重点实验室,兰州730000

第一机构：甘肃中医药大学

年份：2020

卷号：13

期号：1

起止页码：41

中文期刊名：中华骨质疏松和骨矿盐疾病杂志

外文期刊名：CHinese Journal Of Osteoporosis And Bone Mineral Research

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD2019_2020】；

基金：甘肃省高等学校创新能力提升项目(2019A-080);甘肃省高等学校科研项目(2018A-043);敦煌医学与转化省部共建教育部重点实验室开放基金(DHYX17-08);甘肃省高校中(藏)药化学与质量研究省级重点实验室开放基金(zzy-2018-01);甘肃省高校重大疾病分子医学与中医药防治研究重点实验室开放基金(FZYX17-18-11);甘肃省高校大学生就业创业能力提升工程项目(6-1)。

语种：中文

中文关键词：右归丸;膝骨关节炎;凋亡;Ⅱ型胶原

外文关键词：Youguiwan;knee osteoarthritis;apoptosis;collagen Ⅱ

摘要：目的通过观察右归丸对膝骨关节炎(knee osteoarthritis,KOA)模型大鼠软骨组织凋亡和基质降解调控基因的影响,进一步揭示右归丸防治KOA的机制.方法采用数字表法将SD大鼠随机分为假手术对照组,KOA模型组,KOA+硫酸氨基葡萄糖组(硫酸氨基葡萄糖),KOA+右归丸(高、中、低剂量)组,每组10只.采用改良Hulth法制备大鼠KOA模型,分别予相应药物灌胃8周.HE染色法观察软骨组织的形态学变化,并进行Mankin评分;应用实时荧光定量PCR法检测各组大鼠软骨组织B淋巴细胞瘤分子-2(B-cell lymphoma-2,Bcl-2)、B淋巴细胞瘤分子-2相关X蛋白(Bcl-2 associated X protein,Bax)和门冬氨酸特异性半胱氨酸蛋白酶-3(aspartate-specific cysteine protease-3,caspase-3)基因的表达;应用免疫组化法检测各组软骨组织Ⅱ型胶原(collagenⅡ,COLⅡ)的表达.结果假手术组、KOA模型组、KOA+硫酸氨基葡萄糖组、KOA+右归丸高、中、低剂量组的Mankin评分别为0.33±0.073,4.92±0.539,2.76±0.317,3.05±0.253,4.14±0.159,4.38±0.621;COLⅡ评分分别为3.45±0.07,0.39±0.02,3.53±0.11,3.87±0.05,3.15±0.21,0.41±0.13;Bcl-2基因表达分别为2.22±0.08,1.00±0.08,1.75±0.05,1.83±0.11,1.67±0.03,1.12±0.09;Bax的基因表达分别为0.23±0.01,1.00±0.05,0.33±0.03,0.28±0.07,0.42±0.03,0.75±0.01;Caspase-3基因表达分别为0.18±0.01,1.00±0.03,0.26±0.05,0.21±0.02,0.36±0.02,0.83±0.07.与假手术组比较,KOA模型组大鼠软骨组织Mankin评分明显升高,软骨组织Bcl-2基因表达降低,COLⅡ蛋白表达明显降低,Bax和caspase-3基因表达显著升高(P<0.01);KOA模型组关节软骨边缘严重破坏,软骨细胞排列紊乱.与KOA模型组相比,KOA+右归丸高剂量干预大鼠软骨组织Mankin评分明显降低,KOA+右归丸中、高剂量干预大鼠软骨组织Bax和caspase-3的基因表达均明显降低,Bcl-2基因表达和COL-Ⅱ的蛋白表达均明显升高(P<0.05),KOA+右归丸高剂量组软骨结构趋于正常,软骨细胞分布仅偶见不均,关节软骨表面欠光滑.结论右归丸通过上调Bcl-2表达和下调Bax和caspase-3表达减缓COL-Ⅱ的降解,从而有效保护膝骨关节炎模型鼠关节软骨,延缓关节软骨退变.
Objective To observe of articular cartilage in the knee osteoarthritis rats,which was treated by the Youguiwan,and to further reveal its mechanism Methods Sixty SD rats were randomly divided into six groups:a sham control group,a knee osteoarthritis(KOA)model group,a KOA+glucosamine sulfate group,a KOA+Youguiwan(high-dose,middle-dose,and low-dose)group The modified Hulth method was used to make KOA models for 6 weeks The rats were gavaged with corresponding drugs for 8 weeks.Cartibge tissue in each group was observed by HE stai-ning,and it was evaluated by Makin score.The expression of Bcl-2,Bax,and caspase-3 was checked by qRT-PCR.The expression of collagenⅡ(COLⅡ)was analyzed by immunohistochemistry Results Makin score in sham control group,KOA model group,KOA+glucosamine sulfate group,KOA+youguiwan(high-dose,middle-dose and low-dose)group were 0.33±0.073,4.92±0.539,2.76±0.317,3.05±0.253,4.14±0.159,4.38±0.621.COLⅡexpression in above groups were 3 45±0 07,0 39±0 02,3.53±0.11,3.87±0.05,3.15±0.21,0.41±0.13.Bcl-2 gene expression in above groups were 2.22±0.08,1.00±0.08,1.75±0.05,1.83±0.11,1.67±0.03,1.12±0.09.Bax gene expression in above groups were 0.23±0.01,1.00±0.05,0.33±0.03,0.28±0.07,0.42±0.03,0.75±0.01.Caspase-3 gene ex-pression in above groups were 0.18±0.01,1.00±0.03,0.26±0.05,0.21±0.02,0.36±0.02,0.83±0.07.As com-pared with the sham control group,the Makin score was obviously raised in the KOA model group.The gene expression of Bax and caspase-3 was evidently increased in the KOA model group,and the gene expression of Bcl-2 was evidently re-duced in the KOA model group,and the protein expression of COLⅡwas also evidently decreased in the KOA model group(P<0.01),articular cartilage was seriously damaged,and chondrocytes were arranged in disorder.As compared with the KOA model group,the Makin score was declined obviously in the KOA+high-dose Youguiwan group.The gene expression of Bax and caspase-3 was evidently decreased in the KOA+middle-dose,high-dose Youguiwan group,but the gene expression of Bcl-2 was evidently increased in the KOA+middle-dose,high-dose Youguiwan group,and the protein expression of COLⅡwas also evidently raised in the KOA+middle-dose,high-dose Youguiwan group(P<0.05).Carti-lage structure was tended to be normal,the chondrocytes distribution was uneven,and articular cartilage surface was not smooth in the KOA+high-dose Youguiwan group.Conclusion Youguiwan is effective for treating knee osteoarthritis.You-guiwan protects articular cartilage degeneration by up-regulating the expression of rat osteoarthritis cartilage cells Bcl-2,down-regulating the expression of Bax and caspase-3 and reduce the COLⅡdegeneration.