發(fā)布時(shí)間：2018-03-03 05:36

  本文選題：股骨頭缺血壞死　切入點(diǎn)：動(dòng)物模型/幼兔　出處：《遵義醫(yī)學(xué)院》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的檢測(cè)R-spondin1、DKK-1、β-catenin在幼兔激素性股骨頭缺血壞死動(dòng)物模型股骨頭組織中的表達(dá),比較不同劑量甲基潑尼松龍對(duì)人成骨細(xì)胞中R-spondin1的表達(dá)及人成骨細(xì)胞凋亡的影響,探討糖皮質(zhì)激素致幼兔股骨頭缺血壞死的相關(guān)機(jī)制。方法1.幼兔股骨頭壞死組(n=12):激素性股骨頭缺血壞死新西蘭幼兔;對(duì)照組(n=12):正常新西蘭幼兔。應(yīng)用Western Blot、RT-q PCR檢測(cè)方法檢測(cè)Rspondin1、DKK-1、β-catenin在各組股骨頭中的表達(dá)情況,比較各相關(guān)蛋白及基因在各組中的表達(dá)差異,皮爾遜(Pearson)相關(guān)性檢驗(yàn)分析R-spondin1與DKK-1、β-catenin的相關(guān)性。2.培養(yǎng)人成骨細(xì)胞并傳代,激素干預(yù)組:分別加入(10-8mol/L、10-7mol/L、10-6mol/L)甲基潑尼松龍后,繼續(xù)培養(yǎng)72h;空白組:不加甲基潑尼松龍,繼續(xù)培養(yǎng)72h。Western Blot、RT-q PCR檢測(cè)各組R-spondin1的表達(dá)情況,同期檢測(cè)各組的細(xì)胞凋亡率。結(jié)果1.R-spondin1在幼兔股骨頭壞死組與對(duì)照組中的蛋白表達(dá)量分別為0.179±0.028、0.471±0.104,差異具有統(tǒng)計(jì)學(xué)意義(t=9.38,P0.01);β-catenin在幼兔股骨頭壞死組與對(duì)照組中的蛋白表達(dá)量分別為0.195±0.041、0.632±0.129,差異具有統(tǒng)計(jì)學(xué)意義(t=11.15,P0.01);Dkk1在幼兔股骨頭壞死組與對(duì)照組中的蛋白表達(dá)量分別為0.713±0.161、0.181±0.029,差異具有統(tǒng)計(jì)學(xué)意義(t=-11.25,P0.01);在幼兔股骨頭壞死組與對(duì)照組中,R-spondin1與β-catenin的蛋白表達(dá)呈正相關(guān)關(guān)系(r=0.89,P0.01),R-spondin1與Dkk1呈負(fù)相關(guān)關(guān)系(r=-0.945,P0.05)。2.R-spondin1在幼兔股骨頭壞死組與對(duì)照組中的m RNA表達(dá)量分別為0.255±0.039、0.995±0.093,差異具有統(tǒng)計(jì)學(xué)意義(t=25.33,P0.01);β-catenin在幼兔股骨頭壞死組與對(duì)照組中的m RNA表達(dá)量分別為0.216±0.036、1.055±0.083,差異具有統(tǒng)計(jì)學(xué)意義(t=32.49,P0.01);Dkk1在幼兔股骨頭壞死組與對(duì)照組中的m RNA表達(dá)量分別為3.275±0.073、0.986±0.084,差異具有統(tǒng)計(jì)學(xué)意義(t=-10.32,P0.01);在幼兔股骨頭壞死組與對(duì)照組中,R-spondin1與β-catenin的表達(dá)m RNA呈正相關(guān)關(guān)系(r=0.860,P0.01),R-spondin1與Dkk1呈負(fù)相關(guān)關(guān)系(r=-0.945,P0.01)。3.空白組與10-8mol/L、10-7mol/L、10-6mol/L三種不同劑量甲基潑尼松龍干預(yù)培養(yǎng)的細(xì)胞中R-spondin1蛋白表達(dá)量分別為0.622±0.105、0.441±0.125、0.194±0.038、0.112±0.019,R-spondin1蛋白表達(dá)量隨甲基潑尼松龍劑量增大而降低,差異具有統(tǒng)計(jì)學(xué)意義(F=76.37,P0.01);R-spondin1m RNA表達(dá)量分別為0.999±0.168,0.897±0.131,0.624±0.101,0.323±0.033,R-spondin1m RNA表達(dá)量隨甲基潑尼松龍劑量增大而降低,差異具有統(tǒng)計(jì)學(xué)意義(F=64.79,P0.01)。4.空白組與10-8mol/L組、10-7mol/L組、10-6mol/L組的細(xì)胞凋亡率(%)分別為7.41±1.11、13.14±1.92、25.01±5.21、31.99±9.48,細(xì)胞凋亡率隨干預(yù)劑量增大而增大,差異具有統(tǒng)計(jì)學(xué)意義(F=40.80,P0.01)。結(jié)論1.糖皮質(zhì)激素對(duì)人成骨細(xì)胞中R-spondin1的表達(dá)具有負(fù)調(diào)控作用,可能通過負(fù)調(diào)控R-spondin1表達(dá)促進(jìn)人成骨細(xì)胞的凋亡。2.在糖皮質(zhì)激素致幼兔股骨頭缺血壞死模型中,R-spondin1與β-catenin表達(dá)正相關(guān),與DKK-1表達(dá)負(fù)相關(guān),R-spondin1可能負(fù)調(diào)控Wnt/β-catenin信號(hào)通路,促進(jìn)成骨細(xì)胞的凋亡,導(dǎo)致幼兔激素性股骨頭缺血壞死發(fā)生。
[Abstract]:Objective to detect the expression of R-spondin1, DKK-1, beta -catenin in rabbit steroid induced avascular necrosis of femoral head in animal model, comparison of different doses of methylprednisolone on the expression of human R-spondin1 and human bone cells, osteoblast apoptosis, and to explore the mechanism of glucocorticoid induced avascular necrosis of the femoral head in young rabbits methods 1. Rabbits Femoral Head Necrosis group (n=12): steroid induced femoral head necrosis rabbits; the control group (n=12): normal New Zealand rabbits. The application of Western Blot method for detection of RT-q PCR detection of Rspondin1, DKK-1, -catenin beta expression in each group in the femoral head, comparing the difference of the expression of related protein and the genes in each group, Pearson (Pearson) correlation analysis of R-spondin1 and DKK-1 in human osteoblasts and cultured.2. associated beta -catenin, hormone intervention group were added: (10-8mol/L, 10-7mol/L, 10 -6mol/L) methylprednisolone after cultured 72h; control group: without methylprednisolone, cultured for 72h.Western Blot expression was detected by R-spondin1 RT-q PCR, the apoptosis rate of each group. The results of earlier detection of 1.R-spondin1 in rabbit femoral head necrosis group and control group the protein expression were 0.179 + 0.028,0.471 + 0.104, the difference was statistically significant (t=9.38, P0.01); beta -catenin in rabbit femoral head necrosis group and control group the protein expression were 0.195 + 0.041,0.632 + 0.129, the difference was statistically significant (T, =11.15, P0.01); Dkk1 in rabbit femoral head necrosis group and control group the protein expression respectively. 0.713 + 0.161,0.181 + 0.029, the difference was statistically significant (t=-11.25, P0.01); in rabbit femoral head necrosis group and control group, R-spondin1 and beta -catenin protein expression was positively correlated (r=0.89, P0. 01), a negative correlation between R-spondin1 and Dkk1 (r=-0.945, P0.05).2.R-spondin1 in rabbit femoral head necrosis group and control group in M RNA expression were 0.255 + 0.039,0.995 + 0.093, the difference was statistically significant (t=25.33, P0.01); beta -catenin in rabbit femoral head necrosis group and control group. M RNA expression was 0.216 + 0.036,1.055 + 0.083, the difference was statistically significant (t=32.49, P0.01); Dkk1 in rabbit femoral head necrosis group and control group in M RNA expression were 3.275 + 0.073,0.986 + 0.084, the differences were statistically significant (t=-10.32, P0.01); in rabbit femoral head necrosis group and control group, the expression of M RNA was positively related to the relationship between R-spondin1 and beta -catenin (r=0.860, P0.01), a negative correlation between R-spondin1 and Dkk1 (r=-0.945, P0.01).3. control group and 10-8mol/L, 10-7mol/L, 10-6mol and /L in three different doses of methylprednisolone. The expression of R-spondin1 protein were 0.622 + 0.105,0.441 + 0.125,0.194 + 0.038,0.112 + 0.019 support cells, the expression of R-spondin1 protein with methyl prednisolone dose decreased, the difference was statistically significant (F=76.37, P0.01); R-spondin1m RNA expression were 0.999 + 0.168,0.897 + 0.131,0.624 + 0.101,0.323 + 0.033, R-spondin1m expression of RNA with methyl prednisolone dose decreased, the difference was statistically significant (F=64.79, P0.01).4. control group and 10-8mol/L group, 10-7mol/L group, 10-6mol/L group apoptosis rate (%) were 7.41 + 1.11,13.14 + 1.92,25.01 + 5.21,31.99 + 9.48, the apoptosis rate of intervention with the dose increased, the difference was statistically significant (F=40.80, P0.01 1.). Conclusion glucocorticoid negatively regulates human expression of R-spondin1 in osteoblasts, the expression of R-spondin1 may be through negative regulation To promote the apoptosis of.2. cells into bone in glucocorticoid induced avascular necrosis of femoral model young, R-spondin1 expression was positively correlated with beta -catenin, negatively correlated with the expression of DKK-1 and R-spondin1 may negatively regulate Wnt/ beta -catenin signaling pathway to promote cell apoptosis, leading to young rabbits with steroid induced avascular necrosis of femoral head.

【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R726.8

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