MAPK信號(hào)傳導(dǎo)通路的調(diào)控對(duì)皮膚光老化的保護(hù)作用研究
發(fā)布時(shí)間：2018-09-17 07:29
【摘要】：目的研究UVA照射體外培養(yǎng)的皮膚成纖維細(xì)胞后,細(xì)胞形態(tài)變化及相關(guān)細(xì)胞因子變化,檢測(cè)MAPK信號(hào)傳導(dǎo)的始動(dòng)受體EGFR的表達(dá)情況,及MAPK信號(hào)途徑中的下游基因c-jun,c-fos的表達(dá)變化。通過篩選出高效特異的c-junsiRNA后,對(duì)UVA照射后的皮膚成纖維細(xì)胞進(jìn)行轉(zhuǎn)染,探討下調(diào)MAPK下游的基因c-jun對(duì)皮膚光老化中的MMPs,膠原的影響。 方法：1.取年輕成人包皮成纖維細(xì)胞培養(yǎng)傳代,用第5代細(xì)胞做實(shí)驗(yàn)對(duì)象,進(jìn)行細(xì)胞分組,采用不同計(jì)量的UVA(5 J/cm2、10J/cm2、20J/cm2)照射,設(shè)立對(duì)照組(0J/cm2),建立成纖維細(xì)胞光老化模型,培養(yǎng)24h后,光鏡及電鏡下觀察細(xì)胞形態(tài),RT-PCR及酶聯(lián)免疫法(ELISA)測(cè)定照射前后體外培養(yǎng)的皮膚成纖維細(xì)胞的Ⅰ、Ⅲ型前膠原mRNA表達(dá)及蛋白表達(dá)。MTT法測(cè)定細(xì)胞活性。2.采用UVA(5 J/cm210J/cm2,15J/cm2)照射,設(shè)立對(duì)照組,照射后24小時(shí)用realtimePCR檢測(cè)MAPK信號(hào)傳導(dǎo)途徑下游基因c-jun,c-fos,及終末基因MMP-1,MMP-3的mRNA表達(dá)情況。酶聯(lián)免疫法(ELISA)測(cè)定細(xì)胞上清液中的MMP-1,MMP-3表達(dá),weasternblog檢測(cè)c-jun, c-fos的表達(dá)。3.針對(duì)前期實(shí)驗(yàn)中c-jun隨照射劑量依賴性增高,設(shè)計(jì)5條c-junsiRNA, realtimePCR,及westembolg從基因?qū)用婧偷鞍讓用鎸?duì)其篩選,篩選出沉默效率最高的c-junsiRAN,并優(yōu)化轉(zhuǎn)染條件。4。以15J/cm2輻射劑量照射體外培養(yǎng)的成纖維細(xì)胞,2小時(shí)后轉(zhuǎn)染c-junsiRNA,轉(zhuǎn)染后24小時(shí),realtimePCR檢測(cè)MMP-1、MMP-3 mRNA,Ⅰ、Ⅲ型前膠原mRNA,48小時(shí)后酶聯(lián)免疫法(ELISA)檢測(cè)細(xì)胞上清液中MMP-1、MMP-3,Ⅰ、Ⅲ型前膠原含量。5,以(5 J/cm2、10J/cm2、20J/cm2)照射成纖維細(xì)胞,設(shè)立對(duì)照,檢測(cè)MAPK信號(hào)傳導(dǎo)通路的始動(dòng)受體EGFR的表達(dá)情況,檢測(cè)TGF-β1、IGF-1、KGF、VEGF幾個(gè)細(xì)胞因子在照射前后的變化. 結(jié)果：1.隨著UVA照射劑量增加,成纖維細(xì)胞MTT的OD值下降,與對(duì)照組比較,UVA 10 J/cm2、UVA 20 J/cm2照射組OD值明顯下降,有顯著性差異(p0.01)。Ⅰ型(?)mRNA RT-PCR產(chǎn)物積光吸光度隨著UVA照射劑量增加而降低,UVA 10 J/cm2、UVA20J/cm2照射組降低明顯(p0.05,P0.01),UVA 20 J/cm2組抑制膠原合成作用更加明顯,與對(duì)照組及其它兩組比較,有非常顯著性差異(P0.01)。Ⅲ型膠原含量和mRNA表達(dá),與對(duì)照組比較,UVA 20 J/cm2照射組明顯降低,有非常顯著性差異(P0.01)。細(xì)胞上清液中Ⅰ型膠原含量隨著UVA照射劑量增加而降低,呈劑量依賴關(guān)系。UVA 10J/cm2、UVA 20J/cm2照射組,Ⅰ型膠原含量明顯降低,與對(duì)照組比較(P0.05, P0.01),說明成纖維細(xì)胞分別給予UVA 10J/cm2、UVA 20J/cm2照射劑量,均可以抑制膠原合成,且劑量UVA 20 J/cm2組抑制膠原合成作用更加明顯,與對(duì)照組及其它兩組比較,有非常顯著性差異(P0.01)；而5 J/cm2劑量組與對(duì)照組比較,無顯著性差異(P0.05)。Ⅲ型膠原含量,與對(duì)照組比較,UVA 20 J/cm2照射組Ⅲ型膠原含量明顯降低,有非常顯著性差異(P0.01),說明成纖維細(xì)胞在UVA 20J/cm2照射劑量后,能明顯抑制膠原合成,而UVA 5 J/cm2、UVA 10 J/cm2劑量組與對(duì)照組比較,Ⅲ型膠原含量無顯著性差異(P0.05)。2.隨著UVA照射劑量增加,成纖維細(xì)胞c.junmRNA表達(dá)增加,與對(duì)照比較有顯著差異,(p0.01)。c-fosmRNA各組無差異(P0.05)。MMP-1、MMP-3隨著照射劑量增加mRNA表達(dá)也增加,有顯著差異。蛋白表達(dá)也呈現(xiàn)出與基因表達(dá)的一致性。C-jun隨著照射劑量增加而增加,各組間差異p0.01,而c-fos表達(dá)無明顯變化(P0.05)。3,設(shè)計(jì)出5條c-junsiRNA,經(jīng)realtimePCR,及westembolg從基因?qū)用婧偷鞍讓用婀餐瑢?duì)其篩選出沉默效率最高的c-junsiRAN,沉默效率大于80%。序列為：GCAUUCUUGUCACAAUAAATT。4,以15J/cm2輻射劑量照射體外培養(yǎng)的成纖維細(xì)胞,2小時(shí)后轉(zhuǎn)染c-junsiRNA,轉(zhuǎn)染后24小時(shí),MMP-1、MMP-3 mRNA,與對(duì)照組(轉(zhuǎn)染陰性序列組)相比mRNA表達(dá)下調(diào),其中MMP-1(P0.01)、MMP-3(P0.05)。Ⅰ、Ⅲ型前膠原mRNA,相比對(duì)照組(轉(zhuǎn)染陰性序列組)Ⅰ型前膠原(P0.01),Ⅲ型前膠原(P0.05)。48小時(shí)后酶聯(lián)免疫法(ELISA)檢測(cè)細(xì)胞上清液中MMP-1、MMP-3,Ⅰ、Ⅲ型前膠原含量也呈現(xiàn)出與mRNA表達(dá)一致的結(jié)果MMP-1(P0.01)、MMP-3(P0.05)、Ⅰ膠原(P0.01)、Ⅲ型膠原(P0.05)。5.UVA照射體外培養(yǎng)的皮膚成纖維細(xì)胞EGFRmRNA與蛋白表達(dá)呈現(xiàn)一致性,均隨照射劑量增高而增高,5 J/cm2(p0.05),10J/cm2、20J/cm2(P0.01)。UVA照射體外培養(yǎng)的皮膚成纖維細(xì)胞導(dǎo)致TGF-β1、IGF-1、KGF分泌下降, UVA 10 J/cm2、UVA 20 J/cm2照射組明顯下降,與對(duì)照組比較,有顯著性差異(P0.01)。VEGF分泌升高,UVA 20 J/cm2照射組與對(duì)照組比較,有非常顯著性差異(P0.01)。 結(jié)論：1.UVA照射體外培養(yǎng)的皮膚成纖維細(xì)胞,導(dǎo)致細(xì)胞衰老,增殖活性下降；對(duì)Ⅰ型、Ⅲ型膠原合成有抑制作用。2.UVA照射體外培養(yǎng)的皮膚成纖維細(xì)胞激活MAPK信號(hào)傳導(dǎo)通路,其下游基因c-jun上調(diào)明顯,c-fos變化不大,其終末基因MMP-1、MMP-3均上調(diào)。3.經(jīng)realtimePCR,及westernbolg從基因?qū)用婧偷鞍讓用婀餐瑢?duì)5條c-junsiRNA篩選,篩選出沉默效率最高的c-junsiRAN,沉默效率大于80%。序列為：GCAUUCUUGUCACAAUAAATT,名稱為JUN-h-825。4.UVA照射體外培養(yǎng)的成纖維細(xì)胞2小時(shí)后,經(jīng)c-junsiRNA轉(zhuǎn)染后的成纖維細(xì)胞,一定程度上MMP-1,MMP-3可以逆轉(zhuǎn)下調(diào),Ⅰ型、Ⅲ型膠原合成減少的趨勢(shì)也得到減輕。5.UVA照射后體外培養(yǎng)的皮膚成纖維細(xì)胞,EGFR表達(dá)明顯增加,細(xì)胞上清液中的的細(xì)胞因子,TGF-β1、IGF-1.KGF分泌下降；VEGF分泌升高。
[Abstract]:Objective To study the morphological changes of skin fibroblasts cultured in vitro after UVA irradiation and the changes of related cytokines, detect the expression of EGFR, the initiating receptor of MAPK signal transduction, and the expression of c-jun, c-fos, the downstream genes of MAPK signal pathway. Fiber cells were transfected to investigate the effects of down-regulation of c-jun gene downstream of MAPK on MMPs and collagen in skin photoaging.
METHODS: 1. Young adult prepuce fibroblasts were cultured and subcultured. The 5th generation cells were divided into different groups. The control group (0J/cm2) was irradiated with different doses of UVA (5J/cm2, 10J/cm2, 20J/cm2), and the photoaging model of fibroblasts was established. The morphology, RT-PCR and ELISA were observed under light and electron microscopy 24 hours after culture. METHODS: The mRNA expression and protein expression of type I and type III procollagen in cultured skin fibroblasts were measured by ELISA before and after irradiation. The cell viability was measured by MTT assay. 2. UVA (5J/cm210J/cm2, 15J/cm2) was used to irradiate the skin fibroblasts. The control group was set up and the downstream genes of MAPK signal transduction pathway, c-jun, c-fos and terminal gene, MMP-1, were detected by realtime PCR 24 hours after irradiation. MMP-3 mRNA expression. The expression of MMP-1 and MMP-3 in cell supernatant was detected by enzyme-linked immunosorbent assay (ELISA), and the expression of c-jun and c-fos was detected by weasternblog. 3. Five c-junsiRNA, realtime PCR, and westembolg were designed to screen the c-Jun from gene and protein levels. The highest rate of c-junsiRAN was obtained, and the optimal transfection conditions were optimized. 4. The cultured fibroblasts were irradiated with 15J/cm2 radiation, then transfected with c-junsiRNA 2 hours later. MMP-1, MMP-3 mRNA, type I and type III procollagen mRNA were detected by realtime PCR 24 hours after transfection, and MMP-1, MMP-3, type I and type III procollagen mRNA were detected by enzyme-linked immunoassay (ELISA) 48 hours later. Content. 5. Fibroblasts were irradiated with (5J/cm 2, 10J/cm 2, 20J/cm 2) to detect the expression of EGFR, the initiating receptor of MAPK signal transduction pathway, and the changes of TGF-beta 1, IGF-1, KGF and VEGF before and after irradiation.
Results: 1. With the increase of UVA dose, the OD value of MTT of fibroblasts decreased. Compared with the control group, the OD value of UVA 10 J / cm2 and UVA 20 J / cm2 irradiation groups decreased significantly (p0.01). The cumulative absorbance of type I (?) mRNA RT-PCR products decreased with the increase of UVA dose, and UVA 10 J / cm2 and UVA 20 J / cm2 irradiation groups decreased significantly (p0.05, P 0.05). Compared with the control group and the other two groups, the content of collagen type III and the expression of mRNA were significantly lower in the UVA 20 J/cm 2 irradiation group (P 0.01). Compared with the control group (P 0.05, P 0.01), the content of type I collagen in UVA 10J/cm2 and UVA 20J/cm2 irradiation groups decreased significantly, indicating that fibroblasts irradiated with UVA 10J/cm2 and UVA 20J/cm2 could inhibit collagen synthesis, and the effect of UVA 20J/cm2 irradiation on collagen synthesis was more obvious than that of control group (P 0.05, P 0.01). There was a significant difference between the control group and the other two groups (P 0.01), but there was no significant difference between the 5 J / cm 2 dose group and the control group (P 0.05). The content of collagen type III in the UVA 20 J / cm 2 irradiation group was significantly lower than that in the control group (P 0.01), indicating that fibroblasts were significantly lower than that in the UVA 20 J / cm 2 irradiation group (P 0.01). Compared with the control group, the content of collagen type III was not significantly different (P 0.05). 2. With the increase of UVA dose, the expression of C. Jun mRNA in fibroblasts increased significantly (P 0.01). There was no significant difference in the expression of C - fos mRNA between the groups (P 0.05). MMP - 1 and MMP - 3 with the increase of UVA dose. The expression of C-jun increased with the increase of irradiation dose, but the expression of c-fos did not change significantly (P 0.05). 3. Five c-junsiRNA were designed and screened for silencing by realtime PCR and westembolg at both gene and protein levels. The most efficient c-junsiRAN, the silencing efficiency was more than 80%. The sequence was GCAUUCUUGUCACA AAAATT.4, irradiated with 15J/cm 2 radiation dose, transfected with c-junsiRNA 2 hours later, 24 hours after transfection, MMP-1, MMP-3 mRNA expression was down-regulated compared with the control group (transfection negative sequence group), MMP-1 (P 0.01), MMP-3 (P 0.05). MMP-1, MMP-3, type I and type III procollagen in supernatant were detected by enzyme-linked immunosorbent assay (ELISA) 48 hours after transfection. MMP-1 (P 0.01), MMP-3 (P 0.05), type I collagen (P 0.01), type III collagen (P 0.05) and type I procollagen (P 0.05) were also detected by ELISA. 5. The expression of EGFR mRNA and protein in cultured skin fibroblasts irradiated by UVA was consistent with that in vitro. Both EGFR mRNA and protein increased with the increase of irradiation dose. 5 J/cm2 (p0.05), 10 J/cm2, 20 J/cm2 (P 0.01). Cultured skin fibroblasts irradiated by UVA decreased the secretion of TGF-beta 1, IGF-1, KGF, UVA 10 J/cm2 and UVA 20 J/cm2, and decreased significantly compared with the control group. Compared with the control group, there was a significant difference (P 0.01). The secretion of VEGF was elevated. There was a significant difference (P 0.01) between the UVA 20 J/cm 2 irradiation group and the control group.
Conclusion: 1. UVA irradiation of cultured skin fibroblasts can induce cell senescence and decrease proliferation activity, and inhibit the synthesis of collagen type I and type III. 2. UVA irradiation of cultured skin fibroblasts can activate MAPK signal transduction pathway. The downstream genes c-Jun are up-regulated and c-fos is not changed, and the terminal genes MMP-1 and MMP-3 are both up-regulated. Five c-junsiRANs were screened by realtime PCR and Western bolg at the gene and protein levels. The highest silencing efficiency of c-junsiRAN was screened out. The sequence was GCAU UCU UCA CAA UAAATT, named JUN-h-825.4. After 2 hours of UVA irradiation, fibroblasts were transfected with c-junsiRNA. Vitamin cells, to a certain extent, MMP-1 and MMP-3 can reverse the downregulation, and the decrease of collagen synthesis of type I and type III can also be alleviated. 5. After UVA irradiation, the expression of EGFR in cultured skin fibroblasts increased significantly, the secretion of cytokines, TGF-beta 1 and IGF-1.KGF in cell supernatant decreased, and the secretion of VEGF increased.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2011
【分類號(hào)】：R758.1
本文編號(hào)：2245165
[Abstract]:Objective To study the morphological changes of skin fibroblasts cultured in vitro after UVA irradiation and the changes of related cytokines, detect the expression of EGFR, the initiating receptor of MAPK signal transduction, and the expression of c-jun, c-fos, the downstream genes of MAPK signal pathway. Fiber cells were transfected to investigate the effects of down-regulation of c-jun gene downstream of MAPK on MMPs and collagen in skin photoaging.
METHODS: 1. Young adult prepuce fibroblasts were cultured and subcultured. The 5th generation cells were divided into different groups. The control group (0J/cm2) was irradiated with different doses of UVA (5J/cm2, 10J/cm2, 20J/cm2), and the photoaging model of fibroblasts was established. The morphology, RT-PCR and ELISA were observed under light and electron microscopy 24 hours after culture. METHODS: The mRNA expression and protein expression of type I and type III procollagen in cultured skin fibroblasts were measured by ELISA before and after irradiation. The cell viability was measured by MTT assay. 2. UVA (5J/cm210J/cm2, 15J/cm2) was used to irradiate the skin fibroblasts. The control group was set up and the downstream genes of MAPK signal transduction pathway, c-jun, c-fos and terminal gene, MMP-1, were detected by realtime PCR 24 hours after irradiation. MMP-3 mRNA expression. The expression of MMP-1 and MMP-3 in cell supernatant was detected by enzyme-linked immunosorbent assay (ELISA), and the expression of c-jun and c-fos was detected by weasternblog. 3. Five c-junsiRNA, realtime PCR, and westembolg were designed to screen the c-Jun from gene and protein levels. The highest rate of c-junsiRAN was obtained, and the optimal transfection conditions were optimized. 4. The cultured fibroblasts were irradiated with 15J/cm2 radiation, then transfected with c-junsiRNA 2 hours later. MMP-1, MMP-3 mRNA, type I and type III procollagen mRNA were detected by realtime PCR 24 hours after transfection, and MMP-1, MMP-3, type I and type III procollagen mRNA were detected by enzyme-linked immunoassay (ELISA) 48 hours later. Content. 5. Fibroblasts were irradiated with (5J/cm 2, 10J/cm 2, 20J/cm 2) to detect the expression of EGFR, the initiating receptor of MAPK signal transduction pathway, and the changes of TGF-beta 1, IGF-1, KGF and VEGF before and after irradiation.
Results: 1. With the increase of UVA dose, the OD value of MTT of fibroblasts decreased. Compared with the control group, the OD value of UVA 10 J / cm2 and UVA 20 J / cm2 irradiation groups decreased significantly (p0.01). The cumulative absorbance of type I (?) mRNA RT-PCR products decreased with the increase of UVA dose, and UVA 10 J / cm2 and UVA 20 J / cm2 irradiation groups decreased significantly (p0.05, P 0.05). Compared with the control group and the other two groups, the content of collagen type III and the expression of mRNA were significantly lower in the UVA 20 J/cm 2 irradiation group (P 0.01). Compared with the control group (P 0.05, P 0.01), the content of type I collagen in UVA 10J/cm2 and UVA 20J/cm2 irradiation groups decreased significantly, indicating that fibroblasts irradiated with UVA 10J/cm2 and UVA 20J/cm2 could inhibit collagen synthesis, and the effect of UVA 20J/cm2 irradiation on collagen synthesis was more obvious than that of control group (P 0.05, P 0.01). There was a significant difference between the control group and the other two groups (P 0.01), but there was no significant difference between the 5 J / cm 2 dose group and the control group (P 0.05). The content of collagen type III in the UVA 20 J / cm 2 irradiation group was significantly lower than that in the control group (P 0.01), indicating that fibroblasts were significantly lower than that in the UVA 20 J / cm 2 irradiation group (P 0.01). Compared with the control group, the content of collagen type III was not significantly different (P 0.05). 2. With the increase of UVA dose, the expression of C. Jun mRNA in fibroblasts increased significantly (P 0.01). There was no significant difference in the expression of C - fos mRNA between the groups (P 0.05). MMP - 1 and MMP - 3 with the increase of UVA dose. The expression of C-jun increased with the increase of irradiation dose, but the expression of c-fos did not change significantly (P 0.05). 3. Five c-junsiRNA were designed and screened for silencing by realtime PCR and westembolg at both gene and protein levels. The most efficient c-junsiRAN, the silencing efficiency was more than 80%. The sequence was GCAUUCUUGUCACA AAAATT.4, irradiated with 15J/cm 2 radiation dose, transfected with c-junsiRNA 2 hours later, 24 hours after transfection, MMP-1, MMP-3 mRNA expression was down-regulated compared with the control group (transfection negative sequence group), MMP-1 (P 0.01), MMP-3 (P 0.05). MMP-1, MMP-3, type I and type III procollagen in supernatant were detected by enzyme-linked immunosorbent assay (ELISA) 48 hours after transfection. MMP-1 (P 0.01), MMP-3 (P 0.05), type I collagen (P 0.01), type III collagen (P 0.05) and type I procollagen (P 0.05) were also detected by ELISA. 5. The expression of EGFR mRNA and protein in cultured skin fibroblasts irradiated by UVA was consistent with that in vitro. Both EGFR mRNA and protein increased with the increase of irradiation dose. 5 J/cm2 (p0.05), 10 J/cm2, 20 J/cm2 (P 0.01). Cultured skin fibroblasts irradiated by UVA decreased the secretion of TGF-beta 1, IGF-1, KGF, UVA 10 J/cm2 and UVA 20 J/cm2, and decreased significantly compared with the control group. Compared with the control group, there was a significant difference (P 0.01). The secretion of VEGF was elevated. There was a significant difference (P 0.01) between the UVA 20 J/cm 2 irradiation group and the control group.
Conclusion: 1. UVA irradiation of cultured skin fibroblasts can induce cell senescence and decrease proliferation activity, and inhibit the synthesis of collagen type I and type III. 2. UVA irradiation of cultured skin fibroblasts can activate MAPK signal transduction pathway. The downstream genes c-Jun are up-regulated and c-fos is not changed, and the terminal genes MMP-1 and MMP-3 are both up-regulated. Five c-junsiRANs were screened by realtime PCR and Western bolg at the gene and protein levels. The highest silencing efficiency of c-junsiRAN was screened out. The sequence was GCAU UCU UCA CAA UAAATT, named JUN-h-825.4. After 2 hours of UVA irradiation, fibroblasts were transfected with c-junsiRNA. Vitamin cells, to a certain extent, MMP-1 and MMP-3 can reverse the downregulation, and the decrease of collagen synthesis of type I and type III can also be alleviated. 5. After UVA irradiation, the expression of EGFR in cultured skin fibroblasts increased significantly, the secretion of cytokines, TGF-beta 1 and IGF-1.KGF in cell supernatant decreased, and the secretion of VEGF increased.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2011
【分類號(hào)】：R758.1
【參考文獻(xiàn)】
相關(guān)期刊論文 前10條
1 李連喜;胡仁明;;RNA干擾在疾病治療中的研究及應(yīng)用進(jìn)展[J];國(guó)際內(nèi)科學(xué)雜志;2007年07期
2 夏濟(jì)平,畢志剛;皮膚光老化機(jī)制研究進(jìn)展[J];國(guó)外醫(yī)學(xué).皮膚性病學(xué)分冊(cè);2002年05期
3 王小勇,畢志剛;表沒食子兒茶酚沒食子酸酯對(duì)UVA抑制真皮成纖維細(xì)胞膠原合成影響的研究[J];臨床皮膚科雜志;2005年09期
4 姜勇,龔小衛(wèi);MAPK信號(hào)轉(zhuǎn)導(dǎo)通路對(duì)炎癥反應(yīng)的調(diào)控[J];生理學(xué)報(bào);2000年04期
5 倪建華,周華,戴修道;長(zhǎng)波紫外線對(duì)人包皮成纖維細(xì)胞影響的實(shí)驗(yàn)研究[J];實(shí)用預(yù)防醫(yī)學(xué);2001年06期
6 劉垠;劉流;王繼華;魯開化;趙亞南;張曦;伍尚敏;張蔓菁;劉丹;;轉(zhuǎn)化生長(zhǎng)因子β1對(duì)長(zhǎng)波紫外線照射皮膚成纖維細(xì)胞線粒體DNA 4977 bp缺失的影響[J];中國(guó)組織工程研究與臨床康復(fù);2009年50期
7 雷小兵,譚升順,彭振輝;長(zhǎng)波紫外線對(duì)成纖維細(xì)胞膠原合成的影響[J];中國(guó)皮膚性病學(xué)雜志;2000年04期
8 曹宇,湯浩,趙紅艷,張番,楊宇;人皮膚成纖維細(xì)胞在紫外線B照射后的TGF-β和HSP70表達(dá)[J];中國(guó)應(yīng)用生理學(xué)雜志;2002年02期
9 吳瑩,黃愛龍,唐霓,張秉強(qiáng),盧年芳;應(yīng)用RNA干擾治療小鼠急性乙型肝炎病毒感染[J];中華醫(yī)學(xué)雜志;2005年09期
10 王寶璽;;皮膚光老化機(jī)制及其防治研究進(jìn)展[J];中華醫(yī)學(xué)雜志;2007年20期
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