【摘要】：目的：觀察高遷移率族蛋白B1(HMGB1)對(duì)體外人肺動(dòng)脈平滑肌細(xì)胞(HPASMC)增殖、遷移和凋亡的影響。 方法：體外培養(yǎng)人肺動(dòng)脈平滑肌細(xì)胞(HPASMC)至5～8代,然后開始實(shí)驗(yàn)。配制含不同濃度HMGB1 (Ong/ml、1 ng/ml、10 ng/ml、100 ng/ml和1000 ng/ml)的培養(yǎng)液,體外分別培養(yǎng)24小時(shí)和48小時(shí)后,用四甲基偶氮唑藍(lán)(MTT)法檢測(cè)不同濃度的HMGB1對(duì)HPASMC細(xì)胞生長(zhǎng)增殖率的影響。利用Boyden小室法檢測(cè)穿膜細(xì)胞數(shù),觀察含不同濃度HMGB1的無(wú)血清培養(yǎng)基對(duì)HPASMC體外培養(yǎng)20小時(shí)的趨化作用。用TUNEL法檢測(cè)不同濃度HMGB1對(duì)HPASMC作用48小時(shí)后細(xì)胞凋亡情況的影響。 結(jié)果：(1)MTT法檢測(cè)顯示HMGB1對(duì)HPASMC細(xì)胞作用24小時(shí)或48小時(shí)后,與陰性對(duì)照組相比,實(shí)驗(yàn)組細(xì)胞增殖率明顯升高,隨著HMGB1濃度增加,細(xì)胞增殖率升高(P0.01),并在HMGB1濃度為100ng/ml時(shí)達(dá)到平臺(tái),且經(jīng)HMGB1處理48小時(shí)后細(xì)胞增殖率高于24小時(shí)對(duì)應(yīng)濃度組(P0.05)。(2)Boyden小室法趨化實(shí)驗(yàn)顯示隨著HMGB1濃度升高,穿膜細(xì)胞數(shù)增加(P0.01),且HMGB1濃度為100 ng/ml時(shí)趨化作用達(dá)到峰值,HMGB1濃度為1μg/ml時(shí)趨化作用略減弱。(3) TUNEL法檢測(cè)細(xì)胞凋亡結(jié)果顯示,不同濃度HMGB1 (Ong/mL\1 ng/ml、10 ng/ml、100 ng/ml和1000 ng/ml)干預(yù)48小時(shí)后,對(duì)照組和各實(shí)驗(yàn)組的細(xì)胞凋亡率分別為8.56%,8.22%,7.61%,7.50%和7.11%(P0.05)。 結(jié)論：1.HMGB1具有促進(jìn)HPASMC進(jìn)行細(xì)胞增殖和遷移的作用,且該作用對(duì)HMGB1具有濃度依賴性,在HMGB1濃度為100ng/ml時(shí)達(dá)到峰值。 2. HMGB1對(duì)HPASMC的細(xì)胞凋亡無(wú)明顯影響。 3. HMGB1通過(guò)誘導(dǎo)HPASMC增殖和遷移來(lái)促進(jìn)肺血管重構(gòu)。
[Abstract]:Aim: to observe the effects of high mobility group B1 (HMGB1) on proliferation, migration and apoptosis of human pulmonary artery smooth muscle cells (HPASMC) in vitro. Methods: human pulmonary artery smooth muscle cells (HPASMC) were cultured in vitro for 5 ~ 8 passages, and then the experiment was started. The culture medium containing different concentrations of HMGB1 (Ong/ml,1 ng/ml,10 ng/ml,100 ng/ml and 1000 ng/ml) was prepared and cultured in vitro for 24 hours and 48 hours, respectively. The effects of different concentrations of HMGB1 on the growth and proliferation of HPASMC cells were detected by tetramethylazo blue (MTT) assay. Boyden chamber method was used to detect the number of transmembrane cells. The chemotactic effect of serum-free medium containing different concentrations of HMGB1 on HPASMC in vitro for 20 hours was observed. TUNEL assay was used to detect the effect of different concentrations of HMGB1 on apoptosis after 48 hours of HPASMC treatment. Results: (1) MTT assay showed that the proliferation rate of HPASMC cells treated with HMGB1 for 24 hours or 48 hours was significantly higher than that of the negative control group. With the increase of HMGB1 concentration, the cell proliferation rate increased (P0.01). When the concentration of HMGB1 was 100ng/ml, the cell proliferation rate was higher than that of the corresponding concentration group (P0.05). (2) Boyden chamber chemotaxis test showed that with the increase of HMGB1 concentration, the cell proliferation rate after 48 hours of HMGB1 treatment was higher than that of the corresponding concentration group (P0.05). (2). The number of transmembrane cells was increased (P0.01), and the chemotaxis reached the peak when HMGB1 concentration was 100 ng/ml, and the chemotaxis was slightly decreased when HMGB1 concentration was 1 渭 g / ml. (3) the results of TUNEL assay showed that the chemotaxis reached the peak value. After 48 hours of intervention with different concentrations of HMGB1 (Ong/mL\ 1 ng/ml,10 ng/ml,100 ng/ml and 1000 ng/ml), the apoptosis rates of the control group and each experimental group were 8.56%, 8.22%, 7.61%, respectively. 7.50% and 7.11% (P0.05). Conclusion: 1.HMGB1 can promote the proliferation and migration of HPASMC cells, and it has a concentration-dependent effect on HMGB1 and reaches its peak when the concentration of HMGB1 is 100ng/ml. 2. HMGB1 had no effect on the apoptosis of HPASMC. 3. HMGB1 promotes pulmonary vascular remodeling by inducing HPASMC proliferation and migration.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R363

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本文編號(hào)：2432494