SIRT1在骨性關(guān)節(jié)炎滑膜中表達(dá)的相關(guān)研究
發(fā)布時(shí)間：2019-04-01 19:04
【摘要】：目的探討沉默信息調(diào)節(jié)因子2相關(guān)酶1(SIRT1)在骨性關(guān)節(jié)炎(OA)滑膜組織及細(xì)胞中的相關(guān)性表達(dá)。方法①OA組:因膝骨性關(guān)節(jié)炎行全膝關(guān)節(jié)置換術(shù)中所取標(biāo)本共20例,男10例,女10例,年齡62-78歲,中位年齡72.3歲。納入標(biāo)準(zhǔn):符合美國(guó)風(fēng)濕病學(xué)會(huì)2001年推薦OA診斷標(biāo)準(zhǔn);②正常對(duì)照組:因急性外傷致下肢截肢術(shù)或因脛骨平臺(tái)骨折行開放復(fù)位內(nèi)固定術(shù)中所取標(biāo)本10例,男6例,女4例,年齡19-42歲,中位年齡34.2歲。納入保準(zhǔn):術(shù)后病理學(xué)診斷關(guān)節(jié)無病變。標(biāo)本離體后無菌狀態(tài)下放置于裝有生理鹽水的無菌標(biāo)本盒中,2 h內(nèi)送實(shí)驗(yàn)室進(jìn)行滑膜細(xì)胞的分離與培養(yǎng)。用甲苯胺藍(lán)染色對(duì)滑膜細(xì)胞進(jìn)行組織形態(tài)學(xué)觀察,用免疫組化法檢測(cè)滑膜細(xì)胞SIRT1表達(dá)及分布特點(diǎn),用Western blot法檢測(cè)滑膜組織及細(xì)胞中SIRT1表達(dá)。結(jié)果1、甲苯胺藍(lán)染色顯示:滑膜細(xì)胞大部分呈梭形,兩極胞突細(xì)長(zhǎng),末端多與臨近細(xì)胞相連,交織成網(wǎng)狀,網(wǎng)間可見少部分小星型或多邊形細(xì)胞。正常組與OA組滑膜細(xì)胞無明顯形態(tài)學(xué)差異。2、免疫組化結(jié)果顯示:在正常組及OA組滑膜細(xì)胞中SIRT1均可見陽性表達(dá),胞質(zhì)廣泛呈棕黃色,細(xì)胞核及細(xì)胞膜未見明顯黃染,OA組與正常組對(duì)比染色強(qiáng)度下降顯著,兩者之間的差異具有統(tǒng)計(jì)學(xué)意義(t=20.208,P㩳0.01)。3、Western blot法結(jié)果顯示:OA患者滑膜組織及細(xì)胞中SIRT1表達(dá)低于正常組,正常組與OA組灰度值經(jīng)統(tǒng)計(jì)學(xué)分析比較,差異有統(tǒng)計(jì)學(xué)意義(t=7.664,P0.01),滑膜細(xì)胞正常組及OA組灰度值經(jīng)統(tǒng)計(jì)學(xué)分析比較也差異有統(tǒng)計(jì)學(xué)意義(t=8.619,P㩳0.01)。結(jié)論SIRT1與OA導(dǎo)致滑膜炎的發(fā)生發(fā)展密切相關(guān),為SIRT1可以作為OA治療的靶點(diǎn)提供一個(gè)新的理論依據(jù)。
[Abstract]:Aim to investigate the expression of silencing information regulator 2 related enzyme 1 (SIRT1) in synovial tissue and cells of osteoarthritis (OA). Methods 1OA group: 20 cases of total knee arthroplasty due to osteoarthritis of the knee, including 10 males and 10 females, aged 62-78 years (median age 72.3 years), were taken from the patients with osteoarthritis of the knee (n = 20, male 10, female 10, n = 10). Inclusion criteria: meet the OA diagnostic criteria recommended by the American Society of Rheumatology in 2001; 2 normal control group: 10 specimens, 6 males and 4 females, aged 19-42 years (median age 34.2 years), were taken from amputation of lower extremities due to acute trauma or open reduction and internal fixation due to tibial plateau fracture. Inclusion accuracy: postoperative pathological diagnosis of joints without pathological changes. The specimens were placed in an aseptic specimen box containing normal saline in vitro and sent to the laboratory for isolation and culture of synovial cells within 2 hours. The histological morphology of synovial cells was observed by toluidine blue staining. The expression and distribution of SIRT1 in synovial cells were detected by immunohistochemical method, and the expression of SIRT1 in synovial tissues and cells were detected by Western blot method. The results were as follows: 1. Most of the synovial cells were fusiform with long and slender polar processes. The terminal of synovial cells were mostly connected with adjacent cells and interwoven into reticulation. A few small star-shaped or polygonal cells could be seen in the intermeshwork between the synovial cells. There was no significant difference between normal group and OA group in morphology of synovial cells. 2. Immunohistochemical results showed that SIRT1 was expressed in synovial cells in both normal group and OA group, cytoplasm was widely brown, and no obvious yellow staining was found in nucleus and cell membrane, and there was no obvious yellow staining in nucleus and cell membrane of synovial cells in normal group and OA group. The staining intensity of OA group was significantly lower than that of normal group (t = 20.208, P < 0.01). The results of western blot showed that the expression of SIRT1 in synovial tissue and cells of OA patients was lower than that of normal group, and the expression of SIRT1 in synovial tissue and cells of OA patients was significantly lower than that of normal group (P < 0.01). The gray values of normal group and OA group were statistically significant (t = 7.664, P0.01). The gray values of normal group and OA group were also statistically significant (t = 8.619, P < 0.01), and that of normal synovial cell group and OA group was significantly higher than that of normal group (t = 7.664, P0.01). (P < 0.01). Conclusion SIRT1 is closely related to the occurrence and development of synovitis caused by OA, which provides a new theoretical basis for SIRT1 as a target of OA therapy.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R684.3
本文編號(hào)：2451800
[Abstract]:Aim to investigate the expression of silencing information regulator 2 related enzyme 1 (SIRT1) in synovial tissue and cells of osteoarthritis (OA). Methods 1OA group: 20 cases of total knee arthroplasty due to osteoarthritis of the knee, including 10 males and 10 females, aged 62-78 years (median age 72.3 years), were taken from the patients with osteoarthritis of the knee (n = 20, male 10, female 10, n = 10). Inclusion criteria: meet the OA diagnostic criteria recommended by the American Society of Rheumatology in 2001; 2 normal control group: 10 specimens, 6 males and 4 females, aged 19-42 years (median age 34.2 years), were taken from amputation of lower extremities due to acute trauma or open reduction and internal fixation due to tibial plateau fracture. Inclusion accuracy: postoperative pathological diagnosis of joints without pathological changes. The specimens were placed in an aseptic specimen box containing normal saline in vitro and sent to the laboratory for isolation and culture of synovial cells within 2 hours. The histological morphology of synovial cells was observed by toluidine blue staining. The expression and distribution of SIRT1 in synovial cells were detected by immunohistochemical method, and the expression of SIRT1 in synovial tissues and cells were detected by Western blot method. The results were as follows: 1. Most of the synovial cells were fusiform with long and slender polar processes. The terminal of synovial cells were mostly connected with adjacent cells and interwoven into reticulation. A few small star-shaped or polygonal cells could be seen in the intermeshwork between the synovial cells. There was no significant difference between normal group and OA group in morphology of synovial cells. 2. Immunohistochemical results showed that SIRT1 was expressed in synovial cells in both normal group and OA group, cytoplasm was widely brown, and no obvious yellow staining was found in nucleus and cell membrane, and there was no obvious yellow staining in nucleus and cell membrane of synovial cells in normal group and OA group. The staining intensity of OA group was significantly lower than that of normal group (t = 20.208, P < 0.01). The results of western blot showed that the expression of SIRT1 in synovial tissue and cells of OA patients was lower than that of normal group, and the expression of SIRT1 in synovial tissue and cells of OA patients was significantly lower than that of normal group (P < 0.01). The gray values of normal group and OA group were statistically significant (t = 7.664, P0.01). The gray values of normal group and OA group were also statistically significant (t = 8.619, P < 0.01), and that of normal synovial cell group and OA group was significantly higher than that of normal group (t = 7.664, P0.01). (P < 0.01). Conclusion SIRT1 is closely related to the occurrence and development of synovitis caused by OA, which provides a new theoretical basis for SIRT1 as a target of OA therapy.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R684.3
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 呂大鵬;白倫浩;孔冉冉;李毓卓;王勇;;MMP-7、MMP-9及其抑制劑TIMP-3在骨性關(guān)節(jié)炎患者膝關(guān)節(jié)滑膜中的表達(dá)和意義[J];中國(guó)醫(yī)科大學(xué)學(xué)報(bào);2009年04期
，本文編號(hào)：2451800
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