CAJ | 학술논문

目的：观察阿托伐他汀对长期应用皮质激素大鼠骨组织中基质金属蛋白酶-2(matrixmetallo-proteinase-2，MMP-2)、基质金属蛋白酶-9( matrix metalloproteinase-9，MMP-9)及其特异性抑制因子基质金属蛋白酶组织抑制剂-1( tissue inhibitor of matrix metalloproteinases-1，TIMP-1)和基质金属蛋白酶组织抑制剂-2( tissue inhibitor of matrix metalloproteinases-2，TIMP-2) mRNA表达的影响，探讨阿托伐他汀预防激素性股骨头坏死的效果及其作用机制。方法健康SD大鼠30只，采用数字随机法，分为激素组、阿托伐他汀组和对照组3个组，每组10只。激素组和阿托伐他汀组给予肌内注射醋酸泼尼松龙12.5 mg/kg，每周2次；阿托伐他汀组同时给予阿托伐他汀1 mg/kg灌胃，每日1次(按每千克体重最大用量给药，每天最大用量是60 mg，实验动物给药量为60 mg/60 kg体重)；对照组只给予相同体积生理盐水肌注。给药后4周取左侧股骨头骨组织石蜡包埋，HE 染色，鉴定骨质疏松和股骨头坏死情况；取右侧股骨头骨组织提取总 RNA，采用逆转录聚合酶链反应(RT-PCR)技术检测MMP-2、MMP-9、TIMP-1和TIMP-2的mRNA表达水平。结果激素组与阿托伐他汀组各有1只动物死亡。对照组股骨头骨组织切片HE染色可见骨小梁由板层骨构成，绝大多数小梁骨陷窝内可见骨细胞，小梁间为血管和骨髓。激素组表现为骨小梁稀疏和大量不连续的骨碎片及骨髓坏死，碎片骨陷窝内骨细胞大部分消失，周围有大量炎性肉芽组织，阿托伐他汀组介于两者之间，轻度炎性细胞浸润，骨小梁略纤细，多数小梁骨陷窝内可见骨细胞，小梁间为血管和骨髓。MMP-2在激素组、阿托伐他汀组、对照组中的表达分别为0.15±0.04、0.10±0.09、0.09±0.03；MMP-9在3组中的表达分别为0.13±0.03，0.11±0.05和0.08±0.02；TIMP-1在3组中的表达分别为0.07±0.02，0.15±0.05和0.18±0.04；TIMP-2在3组中的表达分别为0.45±0.15，0.73±0.08和0.69±0.19。激素组与对照组比较，MMP-2、MMP-9 mRNA表达增高，TIMP-1、TIMP-2 mRNA的表达降低，差异有统计学意义( P＜0.05)；阿托伐他汀组与对照组比较，除 MMP-9差异有统计学意义外，其余差异无统计学意义；阿托伐他汀组与激素组比较，MMP-2、MMP-9 mRNA表达降低，TIMP-1、TIMP-2mRNA表达增高，差异有统计学意义(P＜0.05)。结论醋酸泼尼松龙上调大鼠骨组织中MMP-2、MMP-9 mRNA的表达水平，下调TIMP-1、TIMP-2的表达水平，使MMPs/TIMPs比值升高。阿托伐他汀可以拮抗皮质激素对MMPs/TIMPs系统的调控。目的：觀察阿託伐他汀對長期應用皮質激素大鼠骨組織中基質金屬蛋白酶-2(matrixmetallo-proteinase-2，MMP-2)、基質金屬蛋白酶-9( matrix metalloproteinase-9，MMP-9)及其特異性抑製因子基質金屬蛋白酶組織抑製劑-1( tissue inhibitor of matrix metalloproteinases-1，TIMP-1)和基質金屬蛋白酶組織抑製劑-2( tissue inhibitor of matrix metalloproteinases-2，TIMP-2) mRNA錶達的影響，探討阿託伐他汀預防激素性股骨頭壞死的效果及其作用機製。方法健康SD大鼠30隻，採用數字隨機法，分為激素組、阿託伐他汀組和對照組3箇組，每組10隻。激素組和阿託伐他汀組給予肌內註射醋痠潑尼鬆龍12.5 mg/kg，每週2次；阿託伐他汀組同時給予阿託伐他汀1 mg/kg灌胃，每日1次(按每韆剋體重最大用量給藥，每天最大用量是60 mg，實驗動物給藥量為60 mg/60 kg體重)；對照組隻給予相同體積生理鹽水肌註。給藥後4週取左側股骨頭骨組織石蠟包埋，HE 染色，鑒定骨質疏鬆和股骨頭壞死情況；取右側股骨頭骨組織提取總 RNA，採用逆轉錄聚閤酶鏈反應(RT-PCR)技術檢測MMP-2、MMP-9、TIMP-1和TIMP-2的mRNA錶達水平。結果激素組與阿託伐他汀組各有1隻動物死亡。對照組股骨頭骨組織切片HE染色可見骨小樑由闆層骨構成，絕大多數小樑骨陷窩內可見骨細胞，小樑間為血管和骨髓。激素組錶現為骨小樑稀疏和大量不連續的骨碎片及骨髓壞死，碎片骨陷窩內骨細胞大部分消失，週圍有大量炎性肉芽組織，阿託伐他汀組介于兩者之間，輕度炎性細胞浸潤，骨小樑略纖細，多數小樑骨陷窩內可見骨細胞，小樑間為血管和骨髓。MMP-2在激素組、阿託伐他汀組、對照組中的錶達分彆為0.15±0.04、0.10±0.09、0.09±0.03；MMP-9在3組中的錶達分彆為0.13±0.03，0.11±0.05和0.08±0.02；TIMP-1在3組中的錶達分彆為0.07±0.02，0.15±0.05和0.18±0.04；TIMP-2在3組中的錶達分彆為0.45±0.15，0.73±0.08和0.69±0.19。激素組與對照組比較，MMP-2、MMP-9 mRNA錶達增高，TIMP-1、TIMP-2 mRNA的錶達降低，差異有統計學意義( P＜0.05)；阿託伐他汀組與對照組比較，除 MMP-9差異有統計學意義外，其餘差異無統計學意義；阿託伐他汀組與激素組比較，MMP-2、MMP-9 mRNA錶達降低，TIMP-1、TIMP-2mRNA錶達增高，差異有統計學意義(P＜0.05)。結論醋痠潑尼鬆龍上調大鼠骨組織中MMP-2、MMP-9 mRNA的錶達水平，下調TIMP-1、TIMP-2的錶達水平，使MMPs/TIMPs比值升高。阿託伐他汀可以拮抗皮質激素對MMPs/TIMPs繫統的調控。
목적：관찰아탁벌타정대장기응용피질격소대서골조직중기질금속단백매-2(matrixmetallo-proteinase-2，MMP-2)、기질금속단백매-9( matrix metalloproteinase-9，MMP-9)급기특이성억제인자기질금속단백매조직억제제-1( tissue inhibitor of matrix metalloproteinases-1，TIMP-1)화기질금속단백매조직억제제-2( tissue inhibitor of matrix metalloproteinases-2，TIMP-2) mRNA표체적영향，탐토아탁벌타정예방격소성고골두배사적효과급기작용궤제。방법건강SD대서30지，채용수자수궤법，분위격소조、아탁벌타정조화대조조3개조，매조10지。격소조화아탁벌타정조급여기내주사작산발니송룡12.5 mg/kg，매주2차；아탁벌타정조동시급여아탁벌타정1 mg/kg관위，매일1차(안매천극체중최대용량급약，매천최대용량시60 mg，실험동물급약량위60 mg/60 kg체중)；대조조지급여상동체적생리염수기주。급약후4주취좌측고골두골조직석사포매，HE 염색，감정골질소송화고골두배사정황；취우측고골두골조직제취총 RNA，채용역전록취합매련반응(RT-PCR)기술검측MMP-2、MMP-9、TIMP-1화TIMP-2적mRNA표체수평。결과격소조여아탁벌타정조각유1지동물사망。대조조고골두골조직절편HE염색가견골소량유판층골구성，절대다수소량골함와내가견골세포，소량간위혈관화골수。격소조표현위골소량희소화대량불련속적골쇄편급골수배사，쇄편골함와내골세포대부분소실，주위유대량염성육아조직，아탁벌타정조개우량자지간，경도염성세포침윤，골소량략섬세，다수소량골함와내가견골세포，소량간위혈관화골수。MMP-2재격소조、아탁벌타정조、대조조중적표체분별위0.15±0.04、0.10±0.09、0.09±0.03；MMP-9재3조중적표체분별위0.13±0.03，0.11±0.05화0.08±0.02；TIMP-1재3조중적표체분별위0.07±0.02，0.15±0.05화0.18±0.04；TIMP-2재3조중적표체분별위0.45±0.15，0.73±0.08화0.69±0.19。격소조여대조조비교，MMP-2、MMP-9 mRNA표체증고，TIMP-1、TIMP-2 mRNA적표체강저，차이유통계학의의( P＜0.05)；아탁벌타정조여대조조비교，제 MMP-9차이유통계학의의외，기여차이무통계학의의；아탁벌타정조여격소조비교，MMP-2、MMP-9 mRNA표체강저，TIMP-1、TIMP-2mRNA표체증고，차이유통계학의의(P＜0.05)。결론작산발니송룡상조대서골조직중MMP-2、MMP-9 mRNA적표체수평，하조TIMP-1、TIMP-2적표체수평，사MMPs/TIMPs비치승고。아탁벌타정가이길항피질격소대MMPs/TIMPs계통적조공。
Objective To investigate the effects of atorvastatin on the mRNA expressions of matrix metalloproteinase-2 ( MMP-2 ), matrix metalloproteinase-9 ( MMP-9 ), tissue inhibitor of matrix metalloproteinases-1 ( TIMP-1 ) and tissue inhibitor of matrix metalloproteinases-2 ( TIMP-2 ) in bone tissues of the rats receiving glucocorticoid for a long time, and to discuss the effects and mechanism of atorvastatin in preventing glucocorticoid-induced femoral head necrosis ( FHN ).Methods A total of 30 healthy adult Sprague-Dawley ( SD ) rats were randomly divided into 3 groups, including glucocorticoid group, atorvastatin group and control group with 10 rats in each group. The rats in the glucocorticoid group and atorvastatin group were treated by intramuscular injection of 12.5 mg / kg prednisolone twice a week. The rats in the atorvastatin group were treated by intragastric administration with 1 mg / kg atorvastatin once a week. The maximum dose was 60 mg each day for experimental animals whose maximum weight was 60 kg. The rats in the control group were treated only by intramuscular injection of the same volume sodium chloride. After 4 weeks’ intervention, the parafifn-embedded osteonecrosis of the left femoral head was detected by HE stain, to identify osteoporosis and FHN. The total Ribonucleic Acid ( RNA ) of the right femoral head was extracted and the mRNA expression levels of MMP-2, MMP-9, TIMP-1 and TIMP-2 mRNAs were examined by reverse transcriptase polymerase chain reaction ( RT-PCR ).Results There was 1 death case in the glucocorticoid group and atorvastatin group respectively. The HE staining of bone tissue slices of the femoral head in the control group showed the bone trabecula was composed of lamellar bone and there were osteocytes in most of the lacunas of the trabecular bone and blood vessels and marrows between bone trabeculae. In the glucocorticoid group, the bony trabecula was sparse and there were a great number of discontinuous bone fragments and marrow necroses. Most of the osteocytes in the lacunas of bone fragments disappeared, with a lot of inlfammatory granulation tissues around. In the atorvastatin group, there was mild inlfammatory cell inifltration and thin bone trabecula. Osteocytes could be seen in most of the lacunas of the trabecular bone, and blood vessels and marrows between bone trabeculae. The expressions of MMP-2 in the glucocorticoid group, atorvastatin group and control group were 0.15±0.04, 0.10±0.09 and 0.09±0.03. The expressions of MMP-9 in 3 groups were 0.13±0.03, 0.11±0.05 and 0.08±0.02. The expressions of TIMP-1 in 3 groups were 0.07±0.02, 0.15±0.05 and 0.18±0.04. The expressions of TIMP-2 in 3 groups were 0.45±0.15, 0.73±0.08 and 0.69±0.19. The mRNA expressions of MMP-2 and MMP-9 in the glucocorticoid group were higher than that in the control group, and while the mRNA expressions of TIMP-1 and TIMP-2 were lower. The differences between them were statistically signiifcant (P<0.05 ). As to the comparison between the atorvastatin group and the control group, statistically signiifcant differences existed only in the expressions of MMP-9. The mRNA expressions of MMP-2 and MMP-9 in the atorvastatin group were lower than that in the control group, and while the mRNA expressions of TIMP-1 and TIMP-2 were higher. The differences between them were statistically signiifcant (P<0.05 ).Conclusions The mRNA expressions of MMP-2 and MMP-9 can be up-regulated and the expressions of TIMP-1 and TIMP-2 can be down-regulated by prednisolone, so as to improve the ratio between MMPs and TIMPs. The glucocorticoid control of MMPs / TIMPs can be antagonized by atorvastatin.