摘要
目的考察丹参酮ⅡA(TSA)微针对瘢痕增生的抑制作用及机制。方法(1)动物实验:将42只新西兰兔随机分成正常组、模型组、空白微针组、阳性药物组(2.5%积雪草总苷)以及丹参酮ⅡA微针(TSA-MN)低、中、高剂量组[(173.93±3.55)μg、(421.44±6.67)μg、(619.94±12.39)μg]。通过新西兰兔耳腹侧面制备创口使其增生,得到兔耳增生性瘢痕模型,连续观察28 d;模型复制成功后给药干预,每日1次,连续21 d。采用HE染色观察增生性瘢痕组织病理形态变化,并测量瘢痕增生指数(SEI)和成纤维细胞密度;采用ELISA法检测兔血清中转化生长因子β1(TGF-β1)、白细胞介素2(IL-2)和白细胞介素4(IL-4)水平。(2)细胞实验:采用体外培养人皮肤成纤维细胞,分为空白对照组、空白微针组、丹参酮ⅡA原料药组(4μg·mL^(-1))以及丹参酮ⅡA微针高、中、低剂量组(6、4、2μg·mL^(-1))。采用Western Blot法检测人皮肤成纤维细胞中TGF-β1、Smad7蛋白表达。结果与正常组比较,模型组兔血清中的TGF-β1、IL-2、IL-4水平及兔耳增生性瘢痕组织SEI值均明显升高(P<0.05)。与模型组比较,丹参酮ⅡA微针中、高剂量组兔血清的IL-2、TGF-β1水平及兔耳增生性瘢痕组织SEI值、成纤维细胞密度均明显降低(P<0.05),丹参酮ⅡA微针高剂量组兔血清的IL-4水平明显降低(P<0.05)。Western Blot实验结果表明,与空白对照组比较,丹参酮ⅡA微针中、高剂量组人皮肤成纤维细胞的TGF-β1蛋白表达明显下调(P<0.05,P<0.01),丹参酮ⅡA微针低、中、高剂量组人皮肤成纤维细胞的Smad7蛋白表达显著上调(P<0.01)。结论丹参酮ⅡA微针有抑制增生性瘢痕的作用,可能与调节TGF-β1/Smad通路及其抗炎作用有关。
Objective To investigate the inhibitory effect and its mechanism of Tanshinone ⅡA(TSA)microneedle on hypertrophic scars(HS). Methods (1)Animal experiment:42 New Zealand rabbits were randomly divided into normal group,model group,blank microneedle group,positive drug group(2.5% asiaticoside)and Tanshinone Ⅱ A microneedle(TSA-MN)low-,medium-and high-dose groups [(173.93±3.55)μg,(421.44±6.67)μg,(619.94±12.39)μg]. A rabbit ear HS model was obtained by preparing a wound on the ventral side of the New Zealand rabbit ear to make it hyperplastic,and the model was observed continuously for 28 days;the intervention was administated once daily for 21 days after successful replication of the model. The histomorphological changes of the hyperplastic scar were observed by HE staining,and the scar proliferation index(SEI) and fibroblast density were measured;the levels of transforming growth factor(TGF-β1),interleukin 2(IL-2)and interleukin 4(IL-4)in rabbit serum were measured by ELISA.(2)Cell experiment:Human skin fibroblasts(HSF)were cultured in vitro and divided into blank control group,blank microneedle group,Tanshinone ⅡA API group(4 μg·mL^(-1))and Tanshinone Ⅱ A microneedle high-,medium-and low-dose groups(6,4 and 2 μg·mL^(-1)). The protein expression of TGF-β1 and Smad7 in human skin fibroblasts was detected by Western Blot method. Results Compared with the normal group,the levels of TGF-β1,IL-2 and IL-4 in serum and the SEI value of ear HS tissue in the model group were significantly increased(P<0.05). Compared with model group, the serum levels of IL-2 and TGF-β1, skin SEI value and fibroblast density of HS tissue were significantly decreased in medium-and high-dose tanshinone Ⅱ A microneedle groups(P<0.05),and the serum IL-4 level was significantly decreased in high-dose tanshinoneⅡ A microneedle group(P<0.05). Western Blot results showed that TGF-β1 protein expression in human skin fibroblasts in mediumand high-dose tanshinoneⅡA microneedle groups was significantly down-regulated compared with blank control group(P<0.05,P<0.01). The expression of Smad7 protein in human skin fibroblasts in low-,medium-and highdose tanshinone Ⅱ A microneedle groups was significantly up-regulated(P<0.01). Conclusion Tanshinone Ⅱ A microneedle has an inhibitory effect on HS,which may be related to modulation of the TGF-β1/Smad pathway and anti-inflammation.
作者
詹燕珊
赖建辉
郭思旖
王纠
韩兵
时军
ZHAN Yanshan;LAI Jianhui;GUO Siyi;WANG Jiu;HAN Bing;SHI Jun(School of Traditional Chinese Medicine,Guangdong Pharmaceutical University,Guangzhou 510006 Guangdong,China;Key Laboratory of Advanced Drug Delivery of Guangdong Province,Guangzhou 510006 Guangdong,China;Plastic Surgery Hospital,Chinese Academy of Medical Sciences and Peking Union Medical,Beijing 100144,China;Guangdong Engineering&Technology Research Center of Precise Drug Delivery System,Guangzhou 510006 Guangdong,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2022年第4期461-467,共7页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金项目(82173982,82103764)。
作者简介
詹燕珊,女,硕士研究生,研究方向:中药制剂研究与开发。Email:2871314142@qq.com。;通信作者:时军,男,博士,副教授,研究方向:中药经皮给药技术研究。Email:shijum8008@163.com。;韩兵,女,博士,医师,研究方向:增生性瘢痕的机制及临床治疗研究。Email:ZXhan8867@163.com。
