Bach1在紫檀芪防御UV致HaCaT細(xì)胞急性光損傷中的作用
發(fā)布時(shí)間：2019-06-22 17:36
【摘要】：[研究背景]皮膚是被覆于體表的人體最大的器官,保護(hù)人體避免遭受各種環(huán)境因素尤其是紫外線(ultraviolet,UV)照射等造成的損傷。UV根據(jù)波長(zhǎng)可分為長(zhǎng)波紫外線(ultraviolet A,UVA,320 - 400nm)、中波紫外線(ultraviolet B,UVB,280 - 320nm)及短波紫外線(ultraviolet C,UVC,200 - 280nm)三類,作用于人體表面的主要是UVA和UVB。長(zhǎng)期或者大量UV輻射可以造成皮膚損傷,如急性日曬傷、慢性光化性皮炎、光老化及皮膚癌等。隨著環(huán)境污染的不斷加重,UV所致皮膚疾病日益嚴(yán)重。目前研究認(rèn)為,UV導(dǎo)致皮膚光損傷的機(jī)制復(fù)雜,除引起DNA雙鏈的破壞和產(chǎn)生嘧啶光產(chǎn)物等直接損傷外,更重要的是氧化應(yīng)激導(dǎo)致過(guò)量的活性氧簇(reactive oxygen species,ROS)堆積,從而對(duì)細(xì)胞的蛋白質(zhì)、DNA、脂質(zhì)等造成損傷,誘發(fā)多種光線性皮膚病。如何清除過(guò)量的ROS、增強(qiáng)抗氧化酶活性、闡明皮膚氧化應(yīng)激防御機(jī)制以及尋找科學(xué)有效的UV防護(hù)劑成為目前的研究熱點(diǎn)。核因子 E2 相關(guān)因子 2 (Nuclear factor erythroid 2-relatedfactor 2,Nrf2 )信號(hào)通路是目前最重要的抗氧化應(yīng)激通路之一。當(dāng)過(guò)量的ROS產(chǎn)生時(shí),Nrf2與Kelch 樣 ECH 聯(lián)合蛋白 1 (kelch-like-Ech-associated - protein 1,Keap1)解偶聯(lián)轉(zhuǎn)位進(jìn)入細(xì)胞核,原本和Maf識(shí)別元件(Maf recognition elements,MAREs)結(jié)合的轉(zhuǎn)錄因子BTB-CNC異體同源體-1 (BTB CNC homology 1, Bach1)解偶聯(lián),胞核內(nèi)Bach1轉(zhuǎn)位至胞漿內(nèi),Nrf2和MAREs形成異二聚體后識(shí)別并結(jié)合抗氧化反應(yīng)元件(antioxidant response element,ARE),調(diào)控下游Ⅱ相解毒酶和抗氧化蛋白的表達(dá),發(fā)揮其抗氧化作用。Bach1之前被認(rèn)為只是同Nrf2競(jìng)爭(zhēng)ARE的結(jié)合位點(diǎn),對(duì)抗氧化基因的表達(dá)起到負(fù)性調(diào)控作用。最近研究發(fā)現(xiàn)Bach1可直接調(diào)控氧化還原、細(xì)胞周期等相關(guān)靶基因的表達(dá),從而在氧化應(yīng)激反應(yīng)以及多種疾病發(fā)生中發(fā)揮重要的調(diào)控作用,可能成為氧化應(yīng)激相關(guān)疾病的新型治療靶點(diǎn)。國(guó)內(nèi)外研究表明,多種天然植物成分如紫檀芪、茶多酚、姜黃素、枸杞多糖、萊菔硫烷、白藜蘆醇等具有抗氧化作用,其中紫檀芪在多種腫瘤細(xì)胞中可通過(guò)調(diào)節(jié)Nrf2信號(hào)通路發(fā)揮抗氧化作用。但紫檀芪對(duì)UV輻照所致皮膚急性光損失是否具有保護(hù)作用以及可能的調(diào)控機(jī)制,目前尚不清楚。因此,本研究擬探討紫檀苗對(duì)UV致人永生化角質(zhì)細(xì)胞(Human keratinocytes,HaCaT)急性光損傷中的防護(hù)作用和Bach1的調(diào)控作用,為新型光防護(hù)劑的開發(fā)提供理論依據(jù)。[目的]1.探討紫檀芪對(duì)UVA/UVB致HaCaT細(xì)胞急性光損傷是否有防護(hù)作用;2.探討紫檀芪對(duì)UVA/UVB致HaCaT細(xì)胞急性光損傷可能防護(hù)機(jī)制;3.探討B(tài)ach1在紫檀芪防御UVA/UVB致HaCaT細(xì)胞急性光損傷中的機(jī)制。[方法]1.分別以 UVA (30 J/cm2)和 UVB (300 mJ/cm2)劑量照射 HaCaT 細(xì)胞,采用MTT法檢測(cè)和倒置顯微鏡觀察照射后的細(xì)胞活性及細(xì)胞形態(tài),以制備HaCaT細(xì)胞急性光損傷模型;2. CCK - 8法和倒置顯微鏡觀察篩選紫檀芪非細(xì)胞毒性作用濃度;3.用MTT法和彗星實(shí)驗(yàn)檢測(cè)紫檀芪對(duì)UVA/UVB照射HaCaT細(xì)胞前后的細(xì)胞活性和DNA損傷情況;4.用WST-1法和DCFH-DA法檢測(cè)紫檀芪對(duì)UVA/UVB照射HaCaT細(xì)胞前后SOD活性和ROS水平的變化;5.用WesternBlot法檢測(cè)紫檀芪對(duì)Nrf2/Bach1蛋白在細(xì)胞質(zhì)和細(xì)胞核中的分布;6.用Western Blot檢測(cè)磷酸激酶抑制劑Genistein對(duì)Nrf2/Bach1蛋白在細(xì)胞質(zhì)和細(xì)胞核中的分布;7.用CCK-8法檢測(cè)磷酸激酶抑制劑Genistein對(duì)UVA/UVB照射紫檀芪孵育HaCaT細(xì)胞前后的細(xì)胞活性變化;8.構(gòu)建過(guò)表達(dá)的干擾短發(fā)夾RNA (short hairpin RNA,shRNA)的慢病毒載體的克隆,將HaCaT細(xì)胞中Nrf2敲低,獲得Nrf2KD細(xì)胞;9.采用qRT-PCR技術(shù)分析HaCaT細(xì)胞和Nrf2KD細(xì)胞中Bach1可能調(diào)控的相關(guān)靶基因。[結(jié)果]1.分別以30 J/cm2的UVA和300 mJ/cm2的UVB劑量照射HaCaT細(xì)胞后其細(xì)胞活性下降(P0.05),細(xì)胞形態(tài)變圓,體積增大,胞內(nèi)顆粒增加。2.用CCK-8法和倒置顯微鏡篩選出紫檀芪的非細(xì)胞毒性作用濃度5μM。3.用MTT法檢測(cè)紫檀芪對(duì)UVA/UVB照射HaCaT細(xì)胞前后的細(xì)胞活性,與對(duì)照相比較,UVA/UVB照射后細(xì)胞活性下降(P0.05);紫檀芪預(yù)處理后細(xì)胞活性上升(P 0.05)。用彗星實(shí)驗(yàn)檢測(cè)紫檀芪對(duì)UVA/UVB照射HaCaT細(xì)胞前后DNA損傷,與對(duì)照相比較,細(xì)胞核的尾長(zhǎng)、尾矩、尾部DNA含量、尾部DNA百分比均顯著增加(P 0.05);紫檀芪預(yù)處理后細(xì)胞核的尾長(zhǎng)、尾矩、尾部DNA含量、尾部DNA百分比均減少(P0.05)。4.用WST-1法檢測(cè)紫檀芪對(duì)UVA/UVB照射HaCaT細(xì)胞前后SOD活性,與對(duì)照組比較,SOD活性上升(P 0.05),紫檀芪處理后SOD活性上升(P0.05);用DCFH-DA法檢測(cè)紫檀芪孵育后HaCaT細(xì)胞ROS水平,與對(duì)照相比較,ROS水平上升(P0.05),紫檀芪預(yù)處理后ROS水平下降(P0.05)。5.用Western Blot法檢測(cè)紫檀芪對(duì)Nrf2/Bach1蛋白在細(xì)胞質(zhì)和細(xì)胞核中的分布,胞核內(nèi)Nrf2增加,Bach1減少。6.用Western Blot法檢測(cè)磷酸酶抑制劑Genistein處理紫檀芪孵育HaCaT細(xì)胞,胞核內(nèi)Bach1增加,Nrf2無(wú)變化。7.用CCK-8法檢測(cè)磷酸激酶抑制劑Genistein對(duì)UVA/UVB照射紫檀芪孵育HaCaT細(xì)胞活性的變化,與對(duì)照組比較,磷酸酶抑制劑Genistein對(duì)HaCaT細(xì)胞的活性無(wú)影響(P0.05),磷酸酶抑制劑Genistein對(duì)UVA/UVB照射紫檀芪孵育HaCaT細(xì)胞的活性下降( 0.05)。8.采用慢病毒將Nrf2-shRNA轉(zhuǎn)染至HaCaT細(xì)胞中,穩(wěn)轉(zhuǎn)細(xì)胞中Nrf2蛋白抑制率80%。9.采用qRT-PCR技術(shù)分析HaCaT細(xì)胞,與未加磷酸酶抑制劑Genistein對(duì)照相比較,bach1、bcl2l1 1、ftl、hmga2、hmox1、mafg、mapt、mmp13、mmp9、sqstm1、tfe3、vrna1-1和vrna1-2基因的表達(dá)上升(P0.05);采用qRT-PCR技術(shù)分析Nrf2KD細(xì)胞,與未加磷酸酶抑制劑Genistein對(duì)照相比較,gclc、gclm、hmga2、hmox1、itpr2、me1和vrnal- 基因的表達(dá)下降(P0.05),mafg、mapt和vegf基因的表達(dá)上升(P0.05)。[結(jié)論]1.紫檀芪對(duì)UVA/UVB致HaCaT細(xì)胞的急性光損傷具有防護(hù)作用;2.紫檀芪對(duì)UVA/UVB致HaCaT細(xì)胞急性光損的防護(hù)作用機(jī)制可能是激活抗氧化信號(hào)通路、提高細(xì)胞活性、降低DNA損傷;3. Bach1在紫檀芪防御UVA/UVB致HaCaT細(xì)胞急性光損傷中起調(diào)控作用,其可能是通過(guò)與Nrf2協(xié)同上調(diào)gclc、gclm、hmga2、hmox1、itpr2、me1和vrna1-2基因的表達(dá),同時(shí)單獨(dú)上調(diào)mafg、mapt和vegf基因的表達(dá)。
[Abstract]:[Study Background] The skin is the largest organ of the human body, which is covered on the body surface, and protects the human body from damage caused by various environmental factors, in particular ultraviolet (UV) irradiation, and the like. The UV wavelength can be divided into three types: long-wave ultraviolet (UVB,320-400 nm), medium-wave ultraviolet (UVB,280-320 nm) and short-wave ultraviolet (ultraviolet C, UVC,200-280 nm). Long-term or large amounts of UV radiation may cause skin lesions, such as acute sunburn, chronic actinic dermatitis, photoaging, and skin cancer. With the increasing of environmental pollution, the skin diseases caused by UV are becoming more and more serious. In the present study, the mechanism of UV-induced skin-light damage is complex, and in addition to the direct injury of the DNA double-chain and the generation of photogenic products, it is more important that oxidative stress causes an excess of reactive oxygen species (ROS) to accumulate, so that the proteins, DNA, The lipid and the like can cause damage and induce a plurality of light-sensitive skin diseases. How to remove excess ROS, enhance the activity of antioxidant enzymes, elucidate the defense mechanism of oxidative stress of the skin, and find a scientific and effective UV protective agent become the current research hotspot. The nuclear factor E2-related factor 2 (Nrf2) signal pathway is one of the most important anti-oxidative stress pathways. When an excess of ROS is produced, Nrf2 is coupled with the Kelch-like-Ech-associated-protein 1, Keap1, to the cell nucleus, and the transcription factor BTB-CNC homologue-1 (BTB CNC homology 1, Bach1), which is bound to the Maf recognition element (MAREs), is de-coupled. Bach1 in the nucleus of the cell is indexed to the cytoplasm, and the Nrf2 and MAREs form heterodimers and then recognize and bind to the anti-oxidation reaction element (ARE) to control the expression of the downstream II-phase detoxification enzyme and the anti-oxidation protein to play an anti-oxidation effect. Bach1 was previously thought to be a binding site to the Nrf2 competition, and a negative regulatory effect on the expression of the anti-oxidation gene. Recent studies have found that Bach1 can directly regulate the expression of related target genes such as redox and cell cycle, so as to play an important regulatory role in oxidative stress reaction and various diseases, and may be a new therapeutic target for oxidative stress-related diseases. The anti-oxidation effect of various natural plant components, such as rosewood, tea polyphenol, curcumin, lycium barbarum polysaccharide, leanthionane, and aloe, is shown at home and abroad, and the anti-oxidation effect can be exerted by adjusting the Nrf2 signal pathway in a variety of tumor cells. However, it is not clear whether the red sandalwood has a protective effect on the acute light loss caused by UV irradiation and the possible regulatory mechanism. Therefore, this study is to explore the protective effect of red sandalwood on the acute light damage of human immortalized keratinocytes (HaCaT) and the regulation of Bach1, and provide a theoretical basis for the development of novel photoprotective agent. [Objective] 1. To investigate the protective effect of red sandalwood on the acute light injury of HaCaT cells induced by UVA/ UVB. Objective To study the possible protective mechanism of red sandalwood on the acute light damage of HaCaT cells induced by UVA/ UVB. To study the mechanism of Bach1 in the treatment of acute light damage to HaCaT cells induced by UVB and UVA/ UVB. [Method] 1. HaCaT cells were irradiated with UVA (30 J/ cm2) and UVB (300 mJ/ cm2) doses, and the cell activity and cell morphology after irradiation were examined by MTT method and inverted microscope to prepare the acute light damage model of HaCaT cells. The non-cytotoxic effect of red sandalwood was observed by CCK-8 and inverted microscope. The cell activity and DNA damage before and after the irradiation of HaCaT cells with UVA/ UVB were detected by MTT and comet assay. The changes of SOD activity and ROS level in red sandalwood were detected by WST-1 method and DCFH-DA method before and after the irradiation of HaCaT cells with UVA/ UVB. The distribution of Nrf2/ Bach1 protein in cytoplasm and nucleus was detected by Western Blot method. The distribution of the Nrf2/ Bach1 protein in the cytoplasm and the nucleus of the phosphokinase inhibitor Genistein was detected by Western Blot. The changes of cell activity before and after the incubation of HaCaT cells with a phosphokinase inhibitor Genistein for UVA/ UVB were detected by the CCK-8 method. The expression of the lentiviral vector with short hairpin RNA (shRNA) was constructed, and the Nrf2 in the HaCaT cell was knocked down to obtain the Nrf2KD cell;9. The related target genes of Bach1 in HaCaT cells and Nrf2KD cells were analyzed by qRT-PCR. [Results] 1. After irradiation of HaCaT cells with UVB dose of 30 J/ cm2 and UVB dose of 300 mJ/ cm2, the cell activity decreased (P0.05). The non-cytotoxic effect of red sandalwood was 5. mu.M.3 by CCK-8 method and inverted microscope. The cell activity before and after the irradiation of HaCaT cells with UVA/ UVB was detected by MTT method, and the cell activity decreased after the irradiation with UVA/ UVB (P0.05); and the cell activity increased after the pre-treatment of the red sandalwood (P 0.05). The DNA damage before and after the irradiation of HaCaT cells with UVA/ UVB was detected by the comet assay, and the tail length, the tail moment, the tail DNA content and the tail DNA percentage of the nucleus were significantly increased (P 0.05), and the tail length, the tail moment and the tail DNA content of the nucleus after the pretreatment of the red sandalwood were significantly increased (P 0.05). The percentage of tail DNA was decreased (P0.05). The activity of SOD in red sandalwood was detected by WST-1 method before and after the irradiation of HaCaT cells with UVA/ UVB. Compared with the control group, the activity of SOD increased (P 0.05), and the activity of SOD in the treatment of the red sandalwood was higher than that of the control group (P 0.05). The level of ROS in HaCaT cells after incubation with DCFH-DA was compared with that of the control group (P0.05). The level of ROS in the pre-treatment of the red sandalwood was decreased (P0.05). The distribution of Nrf2/ Bach1 protein in the cytoplasm and nucleus of the Nrf2/ Bach1 protein was detected by Western Blot method, and the Nrf2 in the nucleus increased and Bach1 decreased. Phosphatase inhibitor Genistein was used to treat the HaCaT cells by Western Blot method, and the Bach1 in the nucleus was increased and Nrf2 was not changed. The changes of the activity of the phosphokinase inhibitor Genistein on the activity of HaCaT cells were detected by using the CCK-8 method. In comparison with the control group, the activity of the phosphatase inhibitor Genistein on the HaCaT cells was not affected (P0.05), and the activity of the phosphatase inhibitor Genistein on the HCaT cells was decreased (0.05). Nrf2-shRNA was transfected into HaCaT cells by lentivirus, and the inhibition rate of Nrf2 protein was 80%. HaCaT cells were analyzed by qRT-PCR, and the expression of bach1, bcl2l1 1, ftl, hmga2, hmox1, mafg, mapi, mp13, mp9, sqstm1, tfe3, vrno1-1 and vrno1-2 was increased (P0.05), and the Nrf2KD cells were analyzed by qRT-PCR. The expression of gclc, gclm, hmga2, hmox1, itpr2, me1, and vrnal-genes decreased (P0.05), and the expression of mafg, mapi, and vegf genes increased (P0.05). [Conclusion] 1. The effect of red sandalwood on the acute light injury of HaCaT cells induced by UVA/ UVB has a protective effect; The protective mechanism of the red sandalwood on the acute light loss of the HaCaT cells induced by UVA/ UVB may be the activation of the anti-oxidation signal pathway, increase the cell activity, and reduce the DNA damage; Bach1 plays an important role in the prevention of acute light damage of HaCaT cells induced by UVA/ UVB, which may be the expression of gclc, gclm, hmga2, hmox1, itpr2, me1, and vrno1-2 by co-regulation with Nrf2, while the expression of mafg, mapi and vegf genes is upregulated.
【學(xué)位授予單位】：廣州醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R758.1
本文編號(hào)：2504816
[Abstract]:[Study Background] The skin is the largest organ of the human body, which is covered on the body surface, and protects the human body from damage caused by various environmental factors, in particular ultraviolet (UV) irradiation, and the like. The UV wavelength can be divided into three types: long-wave ultraviolet (UVB,320-400 nm), medium-wave ultraviolet (UVB,280-320 nm) and short-wave ultraviolet (ultraviolet C, UVC,200-280 nm). Long-term or large amounts of UV radiation may cause skin lesions, such as acute sunburn, chronic actinic dermatitis, photoaging, and skin cancer. With the increasing of environmental pollution, the skin diseases caused by UV are becoming more and more serious. In the present study, the mechanism of UV-induced skin-light damage is complex, and in addition to the direct injury of the DNA double-chain and the generation of photogenic products, it is more important that oxidative stress causes an excess of reactive oxygen species (ROS) to accumulate, so that the proteins, DNA, The lipid and the like can cause damage and induce a plurality of light-sensitive skin diseases. How to remove excess ROS, enhance the activity of antioxidant enzymes, elucidate the defense mechanism of oxidative stress of the skin, and find a scientific and effective UV protective agent become the current research hotspot. The nuclear factor E2-related factor 2 (Nrf2) signal pathway is one of the most important anti-oxidative stress pathways. When an excess of ROS is produced, Nrf2 is coupled with the Kelch-like-Ech-associated-protein 1, Keap1, to the cell nucleus, and the transcription factor BTB-CNC homologue-1 (BTB CNC homology 1, Bach1), which is bound to the Maf recognition element (MAREs), is de-coupled. Bach1 in the nucleus of the cell is indexed to the cytoplasm, and the Nrf2 and MAREs form heterodimers and then recognize and bind to the anti-oxidation reaction element (ARE) to control the expression of the downstream II-phase detoxification enzyme and the anti-oxidation protein to play an anti-oxidation effect. Bach1 was previously thought to be a binding site to the Nrf2 competition, and a negative regulatory effect on the expression of the anti-oxidation gene. Recent studies have found that Bach1 can directly regulate the expression of related target genes such as redox and cell cycle, so as to play an important regulatory role in oxidative stress reaction and various diseases, and may be a new therapeutic target for oxidative stress-related diseases. The anti-oxidation effect of various natural plant components, such as rosewood, tea polyphenol, curcumin, lycium barbarum polysaccharide, leanthionane, and aloe, is shown at home and abroad, and the anti-oxidation effect can be exerted by adjusting the Nrf2 signal pathway in a variety of tumor cells. However, it is not clear whether the red sandalwood has a protective effect on the acute light loss caused by UV irradiation and the possible regulatory mechanism. Therefore, this study is to explore the protective effect of red sandalwood on the acute light damage of human immortalized keratinocytes (HaCaT) and the regulation of Bach1, and provide a theoretical basis for the development of novel photoprotective agent. [Objective] 1. To investigate the protective effect of red sandalwood on the acute light injury of HaCaT cells induced by UVA/ UVB. Objective To study the possible protective mechanism of red sandalwood on the acute light damage of HaCaT cells induced by UVA/ UVB. To study the mechanism of Bach1 in the treatment of acute light damage to HaCaT cells induced by UVB and UVA/ UVB. [Method] 1. HaCaT cells were irradiated with UVA (30 J/ cm2) and UVB (300 mJ/ cm2) doses, and the cell activity and cell morphology after irradiation were examined by MTT method and inverted microscope to prepare the acute light damage model of HaCaT cells. The non-cytotoxic effect of red sandalwood was observed by CCK-8 and inverted microscope. The cell activity and DNA damage before and after the irradiation of HaCaT cells with UVA/ UVB were detected by MTT and comet assay. The changes of SOD activity and ROS level in red sandalwood were detected by WST-1 method and DCFH-DA method before and after the irradiation of HaCaT cells with UVA/ UVB. The distribution of Nrf2/ Bach1 protein in cytoplasm and nucleus was detected by Western Blot method. The distribution of the Nrf2/ Bach1 protein in the cytoplasm and the nucleus of the phosphokinase inhibitor Genistein was detected by Western Blot. The changes of cell activity before and after the incubation of HaCaT cells with a phosphokinase inhibitor Genistein for UVA/ UVB were detected by the CCK-8 method. The expression of the lentiviral vector with short hairpin RNA (shRNA) was constructed, and the Nrf2 in the HaCaT cell was knocked down to obtain the Nrf2KD cell;9. The related target genes of Bach1 in HaCaT cells and Nrf2KD cells were analyzed by qRT-PCR. [Results] 1. After irradiation of HaCaT cells with UVB dose of 30 J/ cm2 and UVB dose of 300 mJ/ cm2, the cell activity decreased (P0.05). The non-cytotoxic effect of red sandalwood was 5. mu.M.3 by CCK-8 method and inverted microscope. The cell activity before and after the irradiation of HaCaT cells with UVA/ UVB was detected by MTT method, and the cell activity decreased after the irradiation with UVA/ UVB (P0.05); and the cell activity increased after the pre-treatment of the red sandalwood (P 0.05). The DNA damage before and after the irradiation of HaCaT cells with UVA/ UVB was detected by the comet assay, and the tail length, the tail moment, the tail DNA content and the tail DNA percentage of the nucleus were significantly increased (P 0.05), and the tail length, the tail moment and the tail DNA content of the nucleus after the pretreatment of the red sandalwood were significantly increased (P 0.05). The percentage of tail DNA was decreased (P0.05). The activity of SOD in red sandalwood was detected by WST-1 method before and after the irradiation of HaCaT cells with UVA/ UVB. Compared with the control group, the activity of SOD increased (P 0.05), and the activity of SOD in the treatment of the red sandalwood was higher than that of the control group (P 0.05). The level of ROS in HaCaT cells after incubation with DCFH-DA was compared with that of the control group (P0.05). The level of ROS in the pre-treatment of the red sandalwood was decreased (P0.05). The distribution of Nrf2/ Bach1 protein in the cytoplasm and nucleus of the Nrf2/ Bach1 protein was detected by Western Blot method, and the Nrf2 in the nucleus increased and Bach1 decreased. Phosphatase inhibitor Genistein was used to treat the HaCaT cells by Western Blot method, and the Bach1 in the nucleus was increased and Nrf2 was not changed. The changes of the activity of the phosphokinase inhibitor Genistein on the activity of HaCaT cells were detected by using the CCK-8 method. In comparison with the control group, the activity of the phosphatase inhibitor Genistein on the HaCaT cells was not affected (P0.05), and the activity of the phosphatase inhibitor Genistein on the HCaT cells was decreased (0.05). Nrf2-shRNA was transfected into HaCaT cells by lentivirus, and the inhibition rate of Nrf2 protein was 80%. HaCaT cells were analyzed by qRT-PCR, and the expression of bach1, bcl2l1 1, ftl, hmga2, hmox1, mafg, mapi, mp13, mp9, sqstm1, tfe3, vrno1-1 and vrno1-2 was increased (P0.05), and the Nrf2KD cells were analyzed by qRT-PCR. The expression of gclc, gclm, hmga2, hmox1, itpr2, me1, and vrnal-genes decreased (P0.05), and the expression of mafg, mapi, and vegf genes increased (P0.05). [Conclusion] 1. The effect of red sandalwood on the acute light injury of HaCaT cells induced by UVA/ UVB has a protective effect; The protective mechanism of the red sandalwood on the acute light loss of the HaCaT cells induced by UVA/ UVB may be the activation of the anti-oxidation signal pathway, increase the cell activity, and reduce the DNA damage; Bach1 plays an important role in the prevention of acute light damage of HaCaT cells induced by UVA/ UVB, which may be the expression of gclc, gclm, hmga2, hmox1, itpr2, me1, and vrno1-2 by co-regulation with Nrf2, while the expression of mafg, mapi and vegf genes is upregulated.
【學(xué)位授予單位】：廣州醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R758.1
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 劉清;江娜;朱慧蘭;;植物來(lái)源的抗氧化劑及其在皮膚科的應(yīng)用[J];皮膚性病診療學(xué)雜志;2015年04期
2 彭世光;閆言;;紫外線對(duì)皮膚成纖維細(xì)胞蛋白差異表達(dá)的影響[J];中國(guó)生物美容;2009年04期
，本文編號(hào)：2504816
本文鏈接：http://sikaile.net/yixuelunwen/pifb/2504816.html
最近更新
教材專著
