两种皮肤细胞对过氧化氢的不同反应及CD147在氧化应激所致人皮肤成纤维细胞衰老中的机制研究
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摘要
第一部分：人皮肤原代角质形成细胞与HaCaT细胞对过氧化氢的不同反应
目的：研究正常人皮肤原代角质形成细胞(NHEKs)与HaCaT细胞株对过氧化氢(H2O2)诱导的氧化应激的不同反应,为选择合适的皮肤细胞模型提供依据。
方法：收集儿童包皮并分离培养原代角质形成细胞,复苏并培养HaCaT细胞,用150uM H2O2处理第2代人皮肤角质形成细胞和HaCaT细胞2小时后继续培养24小时,MTT法检测其细胞生长活性、比色法检测细胞凋亡相关酶caspase-3/7活性、流式细胞仪检测细胞周期中G1期细胞百分数、细胞凋亡百分数及细胞内ROS水平、WST法检测细胞内SOD酶活性水平、细胞衰老相关p-半乳糖苷酶试剂盒检测细胞中p-半乳糖苷酶阳性细胞百分数、Western blot检测衰老相关蛋白Klotho表达水平,并对相关数据进行统计分析。
结果：经H2O2处理后NHEKs与HaCaT细胞活性均明显下降且呈浓度依赖和时间依赖关系,HaCaT细胞的抑制率高于NHEKs(p<0.05), HaCaT细胞内ROS水平、G1期细胞百分数与细胞凋亡相关酶caspase-3/7活性均高于NHEKs细胞,而NHEKs细胞内SOD酶活性水平则高于HaCaT细胞(p<0.05),H2O2处理后两种细胞衰老相关蛋白Klotho表达均下调,NHEKs与HaCaT细胞均均体积变大、圆顿等衰老表型,NHEKs细胞中SA-P-Gal阳性细胞百分数高于对照组(p<0.05),而处理前后HaCaT细胞中均检测不到SA-P-Gal阳性细胞。
结论：NHEKs比HaCaT细胞更能耐受H2O2诱导的氧化应激,细胞经H2O2诱导后,NHEKs既有衰老样的表型,也能表达衰老相关p-半乳糖苷酶,而HaCaT细胞有衰老样的表型,但不表达衰老相关p-半乳糖苷酶。
第二部分CD147对氧化应激所致人皮肤成纤维细胞衰老的影响
目的：观察CD147对H202所致人皮肤成纤维细胞衰老的影响。
方法：收集儿童包皮并分离培养原代成纤维细胞,构建CD147shRNA慢病毒干扰载体,转染第二代成纤维细胞并进行鉴定,100uM H2O2处理转染前后细胞2h,MTT法分别检测诱导后12、24、36、48h细胞生长活性、比色法检测诱导后24h细胞凋亡相关酶caspase-3/7活性、细胞衰老相关p-半乳糖苷酶试剂盒检测诱导后24h细胞中p-半乳糖苷酶活性水平、流式细胞仪检测诱导后24h细胞生长周期中G1期细胞百分数及细胞凋亡率,并对相关数据进行统计分析。结果：观察到干扰组(NHSFs-CD147shRNA组)细胞体积较正常变大、扁平等细胞衰老的形态学表现,其生长活性受到轻度抑制,细胞衰老相关p-半乳糖苷酶、细胞G1期百分数、细胞凋亡相关酶caspase-3/7活性及细胞凋亡率均轻度升高。各细胞组经H202处理后,空载体病毒转染组(NHSFs-virus+H2O2组)细胞较前变大、扁平,少数有多角形等细胞衰老的形态学表现,其细胞生长活性受到中度抑制,细胞衰老相关p-半乳糖苷酶、细胞G1期百分数、凋亡相关酶caspase-3/7活性及细胞凋亡率均中度升高,而CD147干扰组(NHSFs-CD147shRNA+H2O2)细胞体积比对照组明显变大、扁平,并出现多角形等细胞衰老的形态学表现,其生长活性更明显受到抑制。
结论：在正常和氧化应激条件下,CD147对成纤维细胞的衰老均有着一定的调控作用,CD147可能是调控人皮肤细胞衰老的重要分子。
第三部分CD147对氧化应激所致人皮肤成纤维细胞衰老的保护机制研究
目的：探讨CD147对氧化应激所致人皮肤成纤维细胞衰老的可能机制。
方法：收集儿童包皮并分离培养原代成纤维细胞,构建CD147shRNA慢病毒载体并转染第二代成纤维细胞,100uM H2O2处理细胞诱导其衰老,流式细胞仪检测细胞内ROS水平、NBT法检测细胞内SOD活性、Western blot检测处理后细胞衰老相关蛋白Klotho表达水平,并对相关数据进行统计分析。
结果：干扰成纤维细胞中CD147表达后,其细胞内荧光强度稍升高,NHSFs-CD147shRNA组细胞内荧光强度几何均值为7.30±0.58,NHSFs-virus组细胞内荧光强度几何均值为6.87±0.42(P>0.05)；细胞经H2O2处理后,NHSFs-virus+H2O2组细胞内荧光强度几何均值为17.62±1.12,较NHSFs-virus组升高(P<0.05),而NHSFs-CD147shRNA+H2O2组细胞内荧光强度几何均值28.91±1.33,较NHSFs-CD147shRNA组与NHSFs-virus+H2O2组明显升高(P<0.05)。CD147干扰前后,细胞内SOD活性无明显变化,NHSFs-virus组细胞内SOD活性为0.28±0.06U, NHSFs-CD147shRNA组细胞内SOD活性为0.26±0.03U(P>0.05)；细胞经H202处理后,NHSFs-virus+H2O2组细胞内SOD活性为2.18±0.17U,较对照组NHSFs-virus明显升高(P<0.05),NHSFs-CD147shRNA+H2O2组细胞内SOD活性为0.88±0.06U,较对照组NHSFs-CD147shRNA轻度升高(P<0.05),而NHSFS-Virus+H202组细胞内SOD活性较NHSFs-CD147shRNA+H202组高(P<0.05)。细胞经H2O2处理后,NHSFs-virus+H202组细胞衰老相关蛋白Klotho的表达下调,而干扰细胞中CD147的表达后,H202处理前后均检测不到衰老相关蛋白Klotho的表达.
结论：在氧化应激条件下,CD147通过与皮肤成纤维细胞内增加的SOD一起加速对细胞内ROS的清除,减少ROS在细胞内的聚集,从而减轻细胞因氧化应激所致的损伤和衰老,该过程可能与Klotho蛋白相关,CD147可能通过调控细胞衰老相关蛋白Klotho表达而延缓细胞衰老。
Part One Differential Response of Normal Human Epidermal Keratinocytes and HaCaT Cells to Hydrogen Peroxide-induced Oxidative Stress
Objective To investigate the different response of Normal Human Epidermal Keratinocytes and HaCaT cells to hydrogen peroxide-induced oxidative stress.
Methods Normal Human epidermal keratinocytes (NHEKs) were obtained from children's foreskins and HaCaT cells were cultured. We examined changes of cell viability, intracellular reactive oxygen species (ROS) and superoxide dismutase (SOD), caspase-3/7levels, cellular apoptosis, the percent of cells arrested in G1phase, senescence associated β-galactosidase (SA-β-Gal) positive cells and the senescence-related protein klotho in NHEKs and HaCaT cells after H2O2treatment. Statistical analysis was performed using SPSS13.0software.
Results The viability of NHEKs and HaCaT cells decreased in a concentration-dependent and time-dependent manner after exposure to H2O2. The inhibitory effect of150uM H2O2on cell viability was greater in HaCaT cells than in NHEKs (p<0.05). Intracellular ROS, the percent of cells arrested in G1phase and caspase-3/7levels increased more significantly in HaCaT cells than in NHEKs, while intracellular SOD increased more significantly in NHEKs than in HaCaT cells after exposure to150uM H2O2(P<0.05). After H2O2treatment, cells became bigger and uneven in shape, which represents a senescence phenotype. SA β-Gal positive cells increased significantly in NHEKs after treatment with H2O2(P<0.05). Klotho was significantly downregulated in both NHEKs and HaCaT cells after H2O2treatment. However, we didn't observe SA-β-Gal positive HaCaT cells even after treatment with H2O2.
Conclusions NHEKs are more resistant to H2O2-induced oxidative stress than HaCaT cells. HaCaT cells have senescence phenotypes, but do not express p-galactosidase.
Part Two Effects of CD147on Oxidative Stress-induced Senescence in Human Skin Fibroblasts
Objective
To investigate the effects of CD147on senescence of oxidative Stress induced by hydrogen peroxide in human skin fibroblasts.
Methods
Normal human skin fibroblasts (NHSFs) were obtained from children's foreskins. We use lentivirus encoding CD147siRNA to silence the CD147expression in NHSFs. U6-vshRNA-CMV-PUR lentivirus encoding CD147shRNA was constructed. NHSFs were infected with empty lentivirus or lentivirus encoding CD147shRNA, respectively. Cells were treated with100uM H2O2, we examined changes of cell viability with the MTT assay, senescence associated β-galactosidase (SA-β-Gal) positive cells, caspase-3/7levels, cellular apoptosis and the percent of cells arrested in G1phase in NHSFs-virus and NHSFs-CD147shRNA cells and after H2O2treatment or un-treatment. Statistical analysis was performed using SPSS13.0software.
Results
We observed the mild morphological changes related to senescence and associated with a mild increase in the number of SAβ-Gal positive cells in NHSFs-CD147shRNA cells without H2O2exposure, after H2O2treatment, NHSFs-virus cells became bigger and uneven in shape and the percentage of SA P-Gal positive cells increased moderately, representing a senescence phenotype, which was further amplified in NHSFs-CD147shRNA treated cells. The viability of NHSFs-virus treated cells decreased moderately at12,24,36,48h after H2O2treatment, which was further expanded in NHSFs-CD147shRNA cells with H2O2treatment. However, we found that NHSFs-CD147shRNA cells without H2O2exposure can also result in a mild decrease in cell viability. The cells arrested in G1phase resulted in a mild increase in NHSFs-CD147shRNA cells without H2O2exposure, after H2O2treatment, the percentage of cells arrested in G1phase was increased moderately in NHSFs-virus cells, which was further potentiated in NHSFs-CD147shRNA cells. NHSFs-CD147shRNA without H2O2exposure could induce in a mild increase in the caspase-3/7activity, after H2O2treatment, the caspase-3/7activity and the apoptotic rate was increased moderately in NHSFs-virus treated cells, which was further intensified in NHSFs-CD147shRNA cells.
Conclusions
Our results provide a new understanding that CD147may play important roles in protecting NHSFs from senescence under both normal conditions and oxidative stress. The results presented in this study are consistent with the speculation that CD147also plays an regulate-skin aging function.
Part Three The Defense Mechanism Study of against Oxidative Stress-induced Senescence in Human Skin Fibroblasts
Objective
To investigate the defense mechanism of CD147on senescence of oxidative Stress induced by hydrogen peroxide in human skin fibroblasts
Methods
Normal human skin fibroblasts (NHSFs) were obtained from children's foreskins. We use lentivirus encoding CD147siRNA to silence the CD147expression in NHSFs. U6-vshRNA-CMV-PUR lentivirus encoding CD147shRNA was constructed. NHSFs were infected with empty lentivirus or lentivirus encoding CD147shRNA, respectively. Cells were treated with100uM H2O2, we examined changes of intracellular ROS levels, SOD activity and the senescence-related protein klotho in NHSFs-virus and NHSFs-CD147shRNA cells and after H2O2treatment or un-treatment. Statistical analysis was performed using SPSS13.0software.
Results
We observed the intracellular ROS level (Geo Mean) in NHSFs-virus was increased moderately after H2O2treatment (NHSFs-virus+H2O2:17.62±1.12, NHSFs-virus:6.87±0.42, P<0.05), which was further enlarged in NHSFs-CD147shRNA (NHSFs-CD147shRNA+H2O2:28.91±1.33, P<0.05). Whereas NHSFs-CD147shRNA without H2O2exposure can only enhance the intracellular ROS level slightly (NHSFs-CD147shRNA:7.30±0.58, P>0.05). In addition, after H2O2treatment, intracellular SOD level increased moderately in NHSFs-CD147shRNA treated cells (NHSFs-CD147shRNA+H2O2:0.88±0.06, NHSFs-virus:0.28±0.06), which was further augmented in NHSFs-virus cells (NHSFs-virus+H2O2:2.18±0.17, P<0.05). In contrast, CD147suppression alone produced no effect in NHSFs-CD147shRNA cells without H2O2exposure (NHSFs-CD147shRNA:0.26±0.03, P>0.05). However, klotho expression was not detected before or after treatment with H2O2in NHSFs-CD147shRNA.
Conclusions
CD147can exacerbate the oxidative stress-induced senescence induced by H2O2by increasing ROS accumulation and destroying the intrinsic antioxidant defenses in NHSFs. This process may be related to klotho protein. CD147might play a crucial role in anti-oxidation and anti-aging via modulating klotho in NHSFs. We believe that the relationship of CD147and klotho and the exact mechanisms of CD147in senescence of NHSFs might be a novel and potent way to study skin aging.
目的：研究正常人皮肤原代角质形成细胞(NHEKs)与HaCaT细胞株对过氧化氢(H2O2)诱导的氧化应激的不同反应,为选择合适的皮肤细胞模型提供依据。
方法：收集儿童包皮并分离培养原代角质形成细胞,复苏并培养HaCaT细胞,用150uM H2O2处理第2代人皮肤角质形成细胞和HaCaT细胞2小时后继续培养24小时,MTT法检测其细胞生长活性、比色法检测细胞凋亡相关酶caspase-3/7活性、流式细胞仪检测细胞周期中G1期细胞百分数、细胞凋亡百分数及细胞内ROS水平、WST法检测细胞内SOD酶活性水平、细胞衰老相关p-半乳糖苷酶试剂盒检测细胞中p-半乳糖苷酶阳性细胞百分数、Western blot检测衰老相关蛋白Klotho表达水平,并对相关数据进行统计分析。
结果：经H2O2处理后NHEKs与HaCaT细胞活性均明显下降且呈浓度依赖和时间依赖关系,HaCaT细胞的抑制率高于NHEKs(p<0.05), HaCaT细胞内ROS水平、G1期细胞百分数与细胞凋亡相关酶caspase-3/7活性均高于NHEKs细胞,而NHEKs细胞内SOD酶活性水平则高于HaCaT细胞(p<0.05),H2O2处理后两种细胞衰老相关蛋白Klotho表达均下调,NHEKs与HaCaT细胞均均体积变大、圆顿等衰老表型,NHEKs细胞中SA-P-Gal阳性细胞百分数高于对照组(p<0.05),而处理前后HaCaT细胞中均检测不到SA-P-Gal阳性细胞。
结论：NHEKs比HaCaT细胞更能耐受H2O2诱导的氧化应激,细胞经H2O2诱导后,NHEKs既有衰老样的表型,也能表达衰老相关p-半乳糖苷酶,而HaCaT细胞有衰老样的表型,但不表达衰老相关p-半乳糖苷酶。
第二部分CD147对氧化应激所致人皮肤成纤维细胞衰老的影响
目的：观察CD147对H202所致人皮肤成纤维细胞衰老的影响。
方法：收集儿童包皮并分离培养原代成纤维细胞,构建CD147shRNA慢病毒干扰载体,转染第二代成纤维细胞并进行鉴定,100uM H2O2处理转染前后细胞2h,MTT法分别检测诱导后12、24、36、48h细胞生长活性、比色法检测诱导后24h细胞凋亡相关酶caspase-3/7活性、细胞衰老相关p-半乳糖苷酶试剂盒检测诱导后24h细胞中p-半乳糖苷酶活性水平、流式细胞仪检测诱导后24h细胞生长周期中G1期细胞百分数及细胞凋亡率,并对相关数据进行统计分析。结果：观察到干扰组(NHSFs-CD147shRNA组)细胞体积较正常变大、扁平等细胞衰老的形态学表现,其生长活性受到轻度抑制,细胞衰老相关p-半乳糖苷酶、细胞G1期百分数、细胞凋亡相关酶caspase-3/7活性及细胞凋亡率均轻度升高。各细胞组经H202处理后,空载体病毒转染组(NHSFs-virus+H2O2组)细胞较前变大、扁平,少数有多角形等细胞衰老的形态学表现,其细胞生长活性受到中度抑制,细胞衰老相关p-半乳糖苷酶、细胞G1期百分数、凋亡相关酶caspase-3/7活性及细胞凋亡率均中度升高,而CD147干扰组(NHSFs-CD147shRNA+H2O2)细胞体积比对照组明显变大、扁平,并出现多角形等细胞衰老的形态学表现,其生长活性更明显受到抑制。
结论：在正常和氧化应激条件下,CD147对成纤维细胞的衰老均有着一定的调控作用,CD147可能是调控人皮肤细胞衰老的重要分子。
第三部分CD147对氧化应激所致人皮肤成纤维细胞衰老的保护机制研究
目的：探讨CD147对氧化应激所致人皮肤成纤维细胞衰老的可能机制。
方法：收集儿童包皮并分离培养原代成纤维细胞,构建CD147shRNA慢病毒载体并转染第二代成纤维细胞,100uM H2O2处理细胞诱导其衰老,流式细胞仪检测细胞内ROS水平、NBT法检测细胞内SOD活性、Western blot检测处理后细胞衰老相关蛋白Klotho表达水平,并对相关数据进行统计分析。
结果：干扰成纤维细胞中CD147表达后,其细胞内荧光强度稍升高,NHSFs-CD147shRNA组细胞内荧光强度几何均值为7.30±0.58,NHSFs-virus组细胞内荧光强度几何均值为6.87±0.42(P>0.05)；细胞经H2O2处理后,NHSFs-virus+H2O2组细胞内荧光强度几何均值为17.62±1.12,较NHSFs-virus组升高(P<0.05),而NHSFs-CD147shRNA+H2O2组细胞内荧光强度几何均值28.91±1.33,较NHSFs-CD147shRNA组与NHSFs-virus+H2O2组明显升高(P<0.05)。CD147干扰前后,细胞内SOD活性无明显变化,NHSFs-virus组细胞内SOD活性为0.28±0.06U, NHSFs-CD147shRNA组细胞内SOD活性为0.26±0.03U(P>0.05)；细胞经H202处理后,NHSFs-virus+H2O2组细胞内SOD活性为2.18±0.17U,较对照组NHSFs-virus明显升高(P<0.05),NHSFs-CD147shRNA+H2O2组细胞内SOD活性为0.88±0.06U,较对照组NHSFs-CD147shRNA轻度升高(P<0.05),而NHSFS-Virus+H202组细胞内SOD活性较NHSFs-CD147shRNA+H202组高(P<0.05)。细胞经H2O2处理后,NHSFs-virus+H202组细胞衰老相关蛋白Klotho的表达下调,而干扰细胞中CD147的表达后,H202处理前后均检测不到衰老相关蛋白Klotho的表达.
结论：在氧化应激条件下,CD147通过与皮肤成纤维细胞内增加的SOD一起加速对细胞内ROS的清除,减少ROS在细胞内的聚集,从而减轻细胞因氧化应激所致的损伤和衰老,该过程可能与Klotho蛋白相关,CD147可能通过调控细胞衰老相关蛋白Klotho表达而延缓细胞衰老。
Part One Differential Response of Normal Human Epidermal Keratinocytes and HaCaT Cells to Hydrogen Peroxide-induced Oxidative Stress
Objective To investigate the different response of Normal Human Epidermal Keratinocytes and HaCaT cells to hydrogen peroxide-induced oxidative stress.
Methods Normal Human epidermal keratinocytes (NHEKs) were obtained from children's foreskins and HaCaT cells were cultured. We examined changes of cell viability, intracellular reactive oxygen species (ROS) and superoxide dismutase (SOD), caspase-3/7levels, cellular apoptosis, the percent of cells arrested in G1phase, senescence associated β-galactosidase (SA-β-Gal) positive cells and the senescence-related protein klotho in NHEKs and HaCaT cells after H2O2treatment. Statistical analysis was performed using SPSS13.0software.
Results The viability of NHEKs and HaCaT cells decreased in a concentration-dependent and time-dependent manner after exposure to H2O2. The inhibitory effect of150uM H2O2on cell viability was greater in HaCaT cells than in NHEKs (p<0.05). Intracellular ROS, the percent of cells arrested in G1phase and caspase-3/7levels increased more significantly in HaCaT cells than in NHEKs, while intracellular SOD increased more significantly in NHEKs than in HaCaT cells after exposure to150uM H2O2(P<0.05). After H2O2treatment, cells became bigger and uneven in shape, which represents a senescence phenotype. SA β-Gal positive cells increased significantly in NHEKs after treatment with H2O2(P<0.05). Klotho was significantly downregulated in both NHEKs and HaCaT cells after H2O2treatment. However, we didn't observe SA-β-Gal positive HaCaT cells even after treatment with H2O2.
Conclusions NHEKs are more resistant to H2O2-induced oxidative stress than HaCaT cells. HaCaT cells have senescence phenotypes, but do not express p-galactosidase.
Part Two Effects of CD147on Oxidative Stress-induced Senescence in Human Skin Fibroblasts
Objective
To investigate the effects of CD147on senescence of oxidative Stress induced by hydrogen peroxide in human skin fibroblasts.
Methods
Normal human skin fibroblasts (NHSFs) were obtained from children's foreskins. We use lentivirus encoding CD147siRNA to silence the CD147expression in NHSFs. U6-vshRNA-CMV-PUR lentivirus encoding CD147shRNA was constructed. NHSFs were infected with empty lentivirus or lentivirus encoding CD147shRNA, respectively. Cells were treated with100uM H2O2, we examined changes of cell viability with the MTT assay, senescence associated β-galactosidase (SA-β-Gal) positive cells, caspase-3/7levels, cellular apoptosis and the percent of cells arrested in G1phase in NHSFs-virus and NHSFs-CD147shRNA cells and after H2O2treatment or un-treatment. Statistical analysis was performed using SPSS13.0software.
Results
We observed the mild morphological changes related to senescence and associated with a mild increase in the number of SAβ-Gal positive cells in NHSFs-CD147shRNA cells without H2O2exposure, after H2O2treatment, NHSFs-virus cells became bigger and uneven in shape and the percentage of SA P-Gal positive cells increased moderately, representing a senescence phenotype, which was further amplified in NHSFs-CD147shRNA treated cells. The viability of NHSFs-virus treated cells decreased moderately at12,24,36,48h after H2O2treatment, which was further expanded in NHSFs-CD147shRNA cells with H2O2treatment. However, we found that NHSFs-CD147shRNA cells without H2O2exposure can also result in a mild decrease in cell viability. The cells arrested in G1phase resulted in a mild increase in NHSFs-CD147shRNA cells without H2O2exposure, after H2O2treatment, the percentage of cells arrested in G1phase was increased moderately in NHSFs-virus cells, which was further potentiated in NHSFs-CD147shRNA cells. NHSFs-CD147shRNA without H2O2exposure could induce in a mild increase in the caspase-3/7activity, after H2O2treatment, the caspase-3/7activity and the apoptotic rate was increased moderately in NHSFs-virus treated cells, which was further intensified in NHSFs-CD147shRNA cells.
Conclusions
Our results provide a new understanding that CD147may play important roles in protecting NHSFs from senescence under both normal conditions and oxidative stress. The results presented in this study are consistent with the speculation that CD147also plays an regulate-skin aging function.
Part Three The Defense Mechanism Study of against Oxidative Stress-induced Senescence in Human Skin Fibroblasts
Objective
To investigate the defense mechanism of CD147on senescence of oxidative Stress induced by hydrogen peroxide in human skin fibroblasts
Methods
Normal human skin fibroblasts (NHSFs) were obtained from children's foreskins. We use lentivirus encoding CD147siRNA to silence the CD147expression in NHSFs. U6-vshRNA-CMV-PUR lentivirus encoding CD147shRNA was constructed. NHSFs were infected with empty lentivirus or lentivirus encoding CD147shRNA, respectively. Cells were treated with100uM H2O2, we examined changes of intracellular ROS levels, SOD activity and the senescence-related protein klotho in NHSFs-virus and NHSFs-CD147shRNA cells and after H2O2treatment or un-treatment. Statistical analysis was performed using SPSS13.0software.
Results
We observed the intracellular ROS level (Geo Mean) in NHSFs-virus was increased moderately after H2O2treatment (NHSFs-virus+H2O2:17.62±1.12, NHSFs-virus:6.87±0.42, P<0.05), which was further enlarged in NHSFs-CD147shRNA (NHSFs-CD147shRNA+H2O2:28.91±1.33, P<0.05). Whereas NHSFs-CD147shRNA without H2O2exposure can only enhance the intracellular ROS level slightly (NHSFs-CD147shRNA:7.30±0.58, P>0.05). In addition, after H2O2treatment, intracellular SOD level increased moderately in NHSFs-CD147shRNA treated cells (NHSFs-CD147shRNA+H2O2:0.88±0.06, NHSFs-virus:0.28±0.06), which was further augmented in NHSFs-virus cells (NHSFs-virus+H2O2:2.18±0.17, P<0.05). In contrast, CD147suppression alone produced no effect in NHSFs-CD147shRNA cells without H2O2exposure (NHSFs-CD147shRNA:0.26±0.03, P>0.05). However, klotho expression was not detected before or after treatment with H2O2in NHSFs-CD147shRNA.
Conclusions
CD147can exacerbate the oxidative stress-induced senescence induced by H2O2by increasing ROS accumulation and destroying the intrinsic antioxidant defenses in NHSFs. This process may be related to klotho protein. CD147might play a crucial role in anti-oxidation and anti-aging via modulating klotho in NHSFs. We believe that the relationship of CD147and klotho and the exact mechanisms of CD147in senescence of NHSFs might be a novel and potent way to study skin aging.
引文
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