摘要
目的探讨艾灸治疗后豚鼠血清对体外人黑素细胞和角质形成细胞(MC-KC)共培养体系中黑素细胞增殖、凋亡及干细胞生长因子(SCF)、c-kit表达的影响。方法从50只豚鼠中随机选取10只作为空白对照组,其余40只采用随机数字表法按不同观察时间点分为4组,每组10只,分别为T-1组(艾灸治疗后即刻组)、T-2组(艾灸结束后1个月随访组)、T-3组(艾灸结束后2个月随访组)和T-4组(艾灸结束后3个月随访组)。实验前,所有动物背部剃除约2 cm×2 cm区域毛发。T-1组至T-4组豚鼠于经外奇穴“灸癜风”及位于脱毛中心区的“阿是穴”上3 cm处,使用温和灸补法进行悬灸治疗。每次每穴施灸约10 min,每日1次,共治疗1个月。空白对照组仅给予同等时间固定操作,不施灸。比较各组艾灸血清对体外MC-KC共培养体系中人黑素细胞增殖、凋亡,以及c-kit和SCF蛋白及mRNA表达含量的影响。结果细胞增殖实验结果显示,与空白对照组比较,T-1组5%、10%、15%血清的人黑素细胞增殖率均升高,差异均有统计学意义(P<0.05)。与空白对照组比较,T-2组5%、10%血清的人黑素细胞增殖率均升高,差异均有统计学意义(P<0.05)。T-3组5%、10%、15%血清人黑素细胞的增殖与空白对照组比较,差异均无统计学意义(P>0.05)。与空白对照组比较,T-4组15%血清的人黑素细胞增殖率升高,差异有统计学意义(P<0.05)。凋亡实验结果显示,与空白对照组比较,T-1组10%血清能够提高黑素细胞的正常细胞含量,差异有统计学意义(P<0.05)。免疫组织化学和逆转录聚合酶链反应(RT-PCR)实验结果表明,与空白对照组比较,T-1组、T-2组、T-3组和T-4组的艾灸处理组均显著增加人黑素细胞c-kit和SCF蛋白表达,T-1组的艾灸血清能提高c-kit mRNA和SCF mRNA的表达,差异均有统计学意义(P<0.05)。结论艾灸处理能够有效促进体外MC–KC共培养体系中人黑素细胞增殖,并且艾灸血清对黑素细胞c-kit和SCF基因表达及蛋白含量均有显著影响,同时对黑素细胞凋亡具有保护作用。
Objective To investigate the effects of guinea pig serum after moxibustion treatment on the proliferation,apoptosis and stem cell growth factor(SCF),c-kit expression of melanocytes in the co-culture system of human melanocytes and keratinocytes(MC-KC)in vitro.Methods Ten of the 50 guinea pigs were randomly selected as the blank control group,and the remaining 40 were divided into 4 groups according to different observation time points by the random number table method,with 10 in each group,which were T-1 group(immediate group after moxibustion treatment),T-2 group(1 month follow-up group after moxibustion),T-3 group(2 months follow-up group after moxibustion)and T-4 group(3 months follow-up group after moxibustion).A 2 cm×2 cm area of the back of each guinea pig was depilated.Except for the blank control group,the other four groups received moxibustion treatment.The acupoints for moxibustion were the extraordinary acupoint"Jiaobingfeng"and the"Ashi Point,"with the"Ashi Point"positioned within the depilated area.Moxibustion was performed 3 cm above the acupoint for approximately 10 minutes per point,once daily,over the course of one month.The blank control group was only fixed in the same manner without moxibustion.This study compared the effects of moxibustion serum from each group on the proliferation,apoptosis,and expression of c-kit and SCF protein and mRNA content in the in vitro MC-KC co-culture system of human melanocytes.Results The results of cell proliferation assay showed that compared with the blank control group,the proliferation rates of human melanocytes in 5%,10%and 15%serum of T-1 group were increased,and the differences were statistically significant(P<0.05).Compared with the blank control group,the proliferation rates of human melanocytes in 5%and 10%serum of T-2 group were increased,and the differences were statistically significant(P<0.05).There were no statistically significant differences in the proliferation of 5%,10%and 15%serum human melanocytes between the T-3 group and the blank control group(P>0.05).Compared with the blank control group,the proliferation rate of human melanocytes in the 15%serum of the T-4 group was increased,and the differences were statistically significant(P<0.05).The results of melanocyte apoptosis experiment showed that compared with the blank control group,10%serum of T-1 group could increase the normal cell content of melanocytes,and the difference was statistically significant(P<0.05).The results of immunohistochemistry and reverse transcription polymerase chain reaction(RT-PCR)showed that compared with the blank control group,the moxibustion treatment groups of T-1 group,T-2 group,T-3 group and T-4 group significantly increased the expression of c-kit and SCF protein in human melanocytes.The moxibustion serum of T-1 group could increase the expression of c-kit mRNA and SCF mRNA,and the difference was statistically significant(P<0.05).Conclusion The findings demonstrate that moxibustion treatment effectively promotes the proliferation of human melanocytes in the in vitro MC-KC co-culture system.Additionally,moxibustion serum significantly influences the expression of c-kit and SCF genes and proteins in melanocytes,while also offering protection against melanocyte apoptosis.
作者
王帅骅
牛天慧
郭广进
刘石美
王湘宜
成玉
张苍
WANG Shuaihua;NIU Tianhui;GUO Guangjin(Department of Dermatology,Beijing Hospital of Traditional Chinese Medicine,Capital Medical University,Beijing 100010,China;Department of Obstetrics,Gynecology and Reproductive Medicine,Air Force Medical Center,Beijing 100142,China;Aviation Medicine Experimental Center,Air Force Medical Center,Beijing 100142,China)
出处
《临床和实验医学杂志》
2025年第21期2241-2245,共5页
Journal of Clinical and Experimental Medicine
基金
国家自然科学基金项目(编号:82174380)。
