【摘要】：目的:研究胸腺素β4(thymosin beta4)干預(yù)后的人增生性成纖維細(xì)胞(hypertrophicscar fibroblasts HSFB)的基質(zhì)金屬蛋白酶-2(MMP-2)和基質(zhì)金屬蛋白酶(MMP-9)的表達(dá)水平,探討胸腺素β4對(duì)增生性瘢痕可能的作用機(jī)制,提出將胸腺素β4可用于治療增生性瘢痕的理論依據(jù)。方法:選取從我院燒傷科和整形科病人燙傷或燒傷的病人,入選標(biāo)準(zhǔn):1、燒傷半年后,2、瘢痕出現(xiàn)較嚴(yán)重的攣縮,3、未使用其他藥物亦未進(jìn)行其他治療,3、有手術(shù)切除指征并于我院進(jìn)行手術(shù),4、患者和家屬自愿簽署相關(guān)知情同意書(shū)。術(shù)中切除患者瘢痕組織后,將患者瘢痕組織采用組織塊法培養(yǎng)增生性瘢痕成纖維細(xì)胞。取細(xì)胞3-6代細(xì)胞實(shí)驗(yàn),設(shè)對(duì)照組未加入胸腺素β4,其余實(shí)驗(yàn)組分別加入0.05μg/m L,0.1μg/m L,1.0μg/m L,5.0μg/m L的胸腺素β4進(jìn)行干預(yù)24小時(shí)。(1)通過(guò)逆轉(zhuǎn)錄-聚合酶鏈反應(yīng)(RT-PCR)法檢測(cè)胸腺素β4對(duì)HSFB基質(zhì)金屬蛋白酶MMP-2 m RNA和MMP-9 m RNA的表達(dá);(2)通過(guò)免疫印跡法(Western blot)檢測(cè)HSFB基質(zhì)金屬蛋白酶MMP-2和MMP-9蛋白的表達(dá)。結(jié)果:經(jīng)過(guò)24小時(shí)各濃度的胸腺素β4作用后:(1)MMP-2 m RNA和MMP-9 m RNA表達(dá)灰度比值較實(shí)驗(yàn)組均增加,且以5.0μg/m L組增高程度最甚,差異有統(tǒng)計(jì)學(xué)意義(P0.05)(2)MMP-2和MMP-9蛋白表達(dá)灰度比值較對(duì)照組均增加,且以5.0μg/m L組增高程度最甚,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:胸腺素β4能濃度依賴性地增加HSFB中MMP-2,MMP-9因子的表達(dá)。由此可推斷:胸腺素β4可能通過(guò)這種方式來(lái)影響增生性瘢痕的形成。
[Abstract]:Objective: to investigate the expression of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase (MMP-9) in human proliferative fibroblasts (hypertrophicscar fibroblasts HSFB) after intervention of thymosin 尾 4 (thymosin beta4), and to explore the possible mechanism of thymosin 尾 4 on hypertrophic scar. It was suggested that thymosin 尾 4 could be used in the treatment of hypertrophic scar. Methods: the patients who were scalded or burned from the Department of Burn and plastic surgery in our hospital were selected. Standard 1: 1, 2 months after burn, more severe contracture appeared in scar, no other drugs were used and other treatment was not carried out. There were indications of surgical excision and operation in our hospital. Patients and their families voluntarily signed the relevant informed consent. The hypertrophic scar fibroblasts were cultured by tissue mass method after the scar tissue was excised. The 3-6 passage cells were taken from the cells. Thymosin 尾 4 was added to the control group without thymosin 尾 4. The other experimental groups were treated with 0. 0 渭 g / mL Thymosin 尾 4 and 0. 0 渭 g / mL Thymosin 尾 4 for 24 hours. (1) Thymosin 尾 4 was detected by reverse transcription-polymerase chain reaction (RT-PCR) for HSFB matrix metalloproteinase MMP-2 m RNA. (2) the expression of HSFB matrix metalloproteinase MMP-2 and MMP-9 protein was detected by Western blot (Western blot). Results: after 24 hours of thymosin 尾 4 treatment: (1) the gray level ratio of MMP-2 m RNA and MMP-9 m RNA was increased compared with the experimental group, especially in the 5.0 渭 g / mL group. The difference was statistically significant (P0.05) (2) the gray ratio of MMP-2 and MMP-9 protein expression was higher than that of the control group, and the most significant increase was in the 5.0 渭 g / mL group (P0.05). Conclusion: thymosin 尾 4 can increase the expression of MMP-2,MMP-9 factor in HSFB in a concentration dependent manner. It can be concluded that thymosin 尾 4 may affect the formation of hypertrophic scar in this way.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R622

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