【摘要】：【目的】1.建立UVB急性光損傷動(dòng)物模型;2.探究矢車菊素-3葡萄糖(C3G)苷以及納米包被的矢車菊素-3葡萄糖苷(Nano-C3G)對(duì)UVB急性光損傷動(dòng)物模型皮膚的影響并對(duì)比其效果;3.通過p53參與的線粒體細(xì)胞凋亡通路,探究上述影響因素的干預(yù)機(jī)制�！痉椒ā�1.外觀:肉眼觀察評(píng)估;2.皮膚水分及皮膚彈性:多頭皮膚檢測(cè)儀相應(yīng)探頭檢測(cè)皮膚水分以及皮膚彈性;3.組織病理:皮膚組織蘇木素-伊紅(Hematoxylin-Eosin,HE)染色法;皮膚組織TUNEL細(xì)胞凋亡免疫組織組化染色;4.氧化應(yīng)激:硫代巴比妥酸(TBA)法檢測(cè)丙二醛(MDA)、脂質(zhì)過氧化物(LPO);5.DNA損傷:酶聯(lián)免疫吸附實(shí)驗(yàn)(ELISA)法檢測(cè)8羥基脫氧鳥苷(8-OHd G);6.p53:免疫印跡法(Western blotting)檢測(cè)p53表達(dá)水平;7.凋亡相關(guān)蛋白:a)酶聯(lián)免疫吸附實(shí)驗(yàn)(ELISA)法檢測(cè)Bcl-2家族蛋白(Bax、Bcl-2);b)免疫印跡法(Western blotting)檢測(cè)Caspase家族(Caspase-3、Caspase-9);【結(jié)果】1.建立以高UVB劑量照射剃毛法除毛的昆明小鼠,并繼續(xù)飼養(yǎng)24小時(shí)的急性光損傷模型。2.Nano-C3G(125-500u M)能改善UVB急性損傷小鼠皮膚外觀、皮膚水分;TUNEL細(xì)胞凋亡指數(shù)下降;且同等高劑量條件下(500 u M),Nano-C3G藥效較C3G強(qiáng);3.Nano-C3G(125-500u M)能夠有效降低UVB誘導(dǎo)的氧化應(yīng)激產(chǎn)物L(fēng)PO、MDA含量(p0.0001);4.Nano-C3G(125-500u M)能夠有效減少UVB誘導(dǎo)的8-OHd G(p0.0001);且同等高劑量條件下(500 u M),Nano-C3G藥效較C3G強(qiáng)(p0.05);5.Nano-C3G(125-500u M)能夠有效下調(diào)UVB誘導(dǎo)的p53表達(dá)(p0.0001),平衡Bcl-2家族(抗凋亡蛋白Bcl-2、促凋亡蛋白Bax)(p0.0001),下調(diào)Caspase家族(Caspase-3、9)表達(dá)(p0.05);且同等高劑量條件下(500 u M),Nano-C3G藥效較C3G強(qiáng)(p0.05);【結(jié)論】1.UVB照射誘導(dǎo)小鼠皮膚角質(zhì)形成細(xì)胞凋亡,可通過光動(dòng)力性氧化應(yīng)激損傷或非光動(dòng)力性直接損傷DNA,激活p53基因表達(dá),通過調(diào)控Bcl-2家族蛋白等一系列細(xì)胞因子,最終激活Caspase家族以啟動(dòng)凋亡級(jí)聯(lián)反應(yīng);2.Nano-C3G、C3G能對(duì)抗UVB急性光損傷導(dǎo)致的小鼠皮膚表皮組織的細(xì)胞凋亡,且同等濃度下Nano-C3G的能力更強(qiáng);3.Nano-C3G、C3G抑制UVB所導(dǎo)致的細(xì)胞凋亡,可以通過對(duì)抗其中的氧化應(yīng)激反應(yīng)、減少DNA損傷,降低p53基因表達(dá),調(diào)控Bcl-2家族中抗凋亡蛋白Bcl-2、抑凋亡蛋白Bax的表達(dá),減少凋亡相關(guān)細(xì)胞因子的釋放,最終減少Caspase家族表達(dá)實(shí)現(xiàn)。
[Abstract]:[objective] 1. The animal model of UVB acute light injury was established. To investigate the effects of cornulin-3 glucose (C3G) glucoside (C3G) and nano-encapsulated cornulin-3 glucoside (Nano-C3G) on the skin of UVB acute light injury animal model. Through p53 involved in mitochondrial apoptosis pathway, the intervention mechanism of the above factors was explored. [methods] 1. Appearance: naked eye observation and evaluation 2. Skin moisture and skin elasticity: multi-head skin detector to detect skin moisture and skin elasticity. 3. Histopathology: Hematoxylin-eosin HE staining and immunohistochemical staining for TUNEL cell apoptosis. Oxidative stress: malondialdehyde (MDA),) lipid peroxide (LPO) damage was detected by thiobarbituric acid (TBA) assay; enzyme linked immunosorbent assay (ELISA) was used to detect 8-hydroxydeoxyguanosine (8-OHd G) 6.p53; Western blot (Western blotting) was used to detect p53 expression. Apoptosis-related protein: a) Enzyme-linked immunosorbent assay (ELISA) was used to detect Bcl-2 family protein (BaxanBcl-2) and (Western blotting) was used to detect Caspase family (Caspase-3 Caspase-9). [results] 1. Kunming mice with shaved hair were irradiated with high UVB dose and kept for 24 hours. 2. Nano-C3G (125-500u M) could improve the skin appearance of mice with acute UVB injury and decrease the apoptosis index of Tunel cells. At the same high dosage (500 u M) Nano-C3G) than C3G, 3.Nano-C3G (125-500u M) can effectively reduce the content of LPO-MDA induced by UVB (p0.0001) 4.Nano-C3G (125-500u M) can effectively reduce the 8-OHd G (p0.0001) induced by UVB, and the drug efficacy of 500u M) Nano-C3G is better than that of C3G (p0.05) 5.Nano-C3G (125-500u). M) can effectively down-regulate p53 expression induced by UVB (p0.0001), balance Bcl-2 family (anti-apoptotic protein Bcl-2, pro-apoptotic protein Bax) (p0.0001), down-regulate the expression of Caspase family (Caspase-3O9) (p0.05), and at the same high dosage (500 u M) + Nano-C3G), the drug efficacy of 1.UVB is better than that of C3G; [conclusion] 1.UVB irradiation is less effective than C3G. [conclusion] Apoptosis of keratinocytes in rat skin The expression of p53 gene can be activated by photodynamic oxidative stress injury or non-photodynamic direct damage of DNA, and a series of cytokines such as Bcl-2 family proteins can be regulated. Finally, the Caspase family was activated to initiate the apoptotic cascade reaction. 2. Nano-C3GN C3G could antagonize the apoptosis of mouse skin epidermis induced by acute light injury of UVB, and the ability of Nano-C3G at the same concentration was stronger. 3. Nano-C3GN C3G inhibited the apoptosis induced by UVB. It can reduce DNA damage, reduce p53 gene expression, regulate the expression of anti-apoptotic protein Bcl-2, inhibit the expression of apoptotic protein Bax, and reduce the release of apoptosis-related cytokines by antagonizing the oxidative stress reaction, reducing the expression of p53 gene, regulating the expression of anti-apoptotic protein Bcl-2 in the Bcl-2 family. Finally, the realization of Caspase family expression is reduced.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R275.9

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