中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2011年
10期
693-696
,共4页
硬皮病,系统性%成纤维细胞%克隆细胞%胶原Ⅰ型%丹参
硬皮病,繫統性%成纖維細胞%剋隆細胞%膠原Ⅰ型%丹參
경피병,계통성%성섬유세포%극륭세포%효원Ⅰ형%단삼
Scleroderma,systemic%Fibroblasts%Clone cells%Collagen type Ⅰ%Salvia miltiorrhiza
目的 探讨丹参对系统性硬化病患者皮损高胶原合成成纤维细胞克隆Ⅰ型前胶原基因转录的调控.方法 采用系统性硬化病患者皮损和正常人皮肤胶原合成异质性成纤维细胞克隆,通过四甲基偶氮唑盐比色法,检测1 g/L丹参注射液及其主要水溶性单体(20 mg/L丹参素钠、5 mg/L丹酚酸B、5 mg/L原儿茶醛)和脂溶性单体(5 mg/L丹参酮ⅡA)对系统性硬化病和正常人高、低胶原合成克隆增殖活性(A490)的影响.利用瞬时转染、双荧光素酶报告基因技术,检测上述药物对克隆Ⅰ型前胶原α1链(COL1A1)基因近端启动区活性的调控.结果 在早期(≤3 d),丹参注射液及单体对成纤维细胞克隆增殖的抑制不显著(P＞0.05).但随作用时间延长,它们对克隆增殖的抑制作用多逐渐增强,第5天,丹参注射液组与水溶性阴性对照组比较,差异有统计学意义(q′=3.22,P＜0.01)；第7天,丹参注射液组、丹酚酸B组和原儿茶醛组与水溶性阴性对照组比较,差异均有统计学意义(q′分别为4.74、3.03、2.56,P值均＜0.05).第5天和第7天,丹参酮ⅡA组与脂溶性阴性对照组比较,差异均有统计学意义(t值分别为2.22 和2.15,P值均＜0.05).丹参注射液、丹参酮ⅡA和原儿茶醛抑制系统性硬化病和正常人成纤维细胞克隆中COL1A1近端启动区的活性(P＜0.01),前两种药优先下调系统性硬化病高胶原合成克隆中COL1A1 近端启动区的活性,COL1A1近端启动区在系统性硬化病高、低胶原合成克隆中的活性水溶性阴性对照组为12.019±0.830和5.388±0.480,丹参注射液组为4.445±1.061和2.856±0.597,F=31.78,P＜0.01；脂溶性阴性对照组为14.155±0.672和4.299±0.252,丹参酮ⅡA组为9.638±0.854和3.192±0.450,F=24.10,P＜0.01.结论 丹参可抑制系统性硬化病高胶原合成成纤维细胞克隆Ⅰ型前胶原基因的转录,其单体丹参酮ⅡA和原儿茶醛可能发挥主要作用.
目的 探討丹參對繫統性硬化病患者皮損高膠原閤成成纖維細胞剋隆Ⅰ型前膠原基因轉錄的調控.方法 採用繫統性硬化病患者皮損和正常人皮膚膠原閤成異質性成纖維細胞剋隆,通過四甲基偶氮唑鹽比色法,檢測1 g/L丹參註射液及其主要水溶性單體(20 mg/L丹參素鈉、5 mg/L丹酚痠B、5 mg/L原兒茶醛)和脂溶性單體(5 mg/L丹參酮ⅡA)對繫統性硬化病和正常人高、低膠原閤成剋隆增殖活性(A490)的影響.利用瞬時轉染、雙熒光素酶報告基因技術,檢測上述藥物對剋隆Ⅰ型前膠原α1鏈(COL1A1)基因近耑啟動區活性的調控.結果 在早期(≤3 d),丹參註射液及單體對成纖維細胞剋隆增殖的抑製不顯著(P＞0.05).但隨作用時間延長,它們對剋隆增殖的抑製作用多逐漸增彊,第5天,丹參註射液組與水溶性陰性對照組比較,差異有統計學意義(q′=3.22,P＜0.01)；第7天,丹參註射液組、丹酚痠B組和原兒茶醛組與水溶性陰性對照組比較,差異均有統計學意義(q′分彆為4.74、3.03、2.56,P值均＜0.05).第5天和第7天,丹參酮ⅡA組與脂溶性陰性對照組比較,差異均有統計學意義(t值分彆為2.22 和2.15,P值均＜0.05).丹參註射液、丹參酮ⅡA和原兒茶醛抑製繫統性硬化病和正常人成纖維細胞剋隆中COL1A1近耑啟動區的活性(P＜0.01),前兩種藥優先下調繫統性硬化病高膠原閤成剋隆中COL1A1 近耑啟動區的活性,COL1A1近耑啟動區在繫統性硬化病高、低膠原閤成剋隆中的活性水溶性陰性對照組為12.019±0.830和5.388±0.480,丹參註射液組為4.445±1.061和2.856±0.597,F=31.78,P＜0.01；脂溶性陰性對照組為14.155±0.672和4.299±0.252,丹參酮ⅡA組為9.638±0.854和3.192±0.450,F=24.10,P＜0.01.結論 丹參可抑製繫統性硬化病高膠原閤成成纖維細胞剋隆Ⅰ型前膠原基因的轉錄,其單體丹參酮ⅡA和原兒茶醛可能髮揮主要作用.
목적 탐토단삼대계통성경화병환자피손고효원합성성섬유세포극륭Ⅰ형전효원기인전록적조공.방법 채용계통성경화병환자피손화정상인피부효원합성이질성성섬유세포극륭,통과사갑기우담서염비색법,검측1 g/L단삼주사액급기주요수용성단체(20 mg/L단삼소납、5 mg/L단분산B、5 mg/L원인다철)화지용성단체(5 mg/L단삼동ⅡA)대계통성경화병화정상인고、저효원합성극륭증식활성(A490)적영향.이용순시전염、쌍형광소매보고기인기술,검측상술약물대극륭Ⅰ형전효원α1련(COL1A1)기인근단계동구활성적조공.결과 재조기(≤3 d),단삼주사액급단체대성섬유세포극륭증식적억제불현저(P＞0.05).단수작용시간연장,타문대극륭증식적억제작용다축점증강,제5천,단삼주사액조여수용성음성대조조비교,차이유통계학의의(q′=3.22,P＜0.01)；제7천,단삼주사액조、단분산B조화원인다철조여수용성음성대조조비교,차이균유통계학의의(q′분별위4.74、3.03、2.56,P치균＜0.05).제5천화제7천,단삼동ⅡA조여지용성음성대조조비교,차이균유통계학의의(t치분별위2.22 화2.15,P치균＜0.05).단삼주사액、단삼동ⅡA화원인다철억제계통성경화병화정상인성섬유세포극륭중COL1A1근단계동구적활성(P＜0.01),전량충약우선하조계통성경화병고효원합성극륭중COL1A1 근단계동구적활성,COL1A1근단계동구재계통성경화병고、저효원합성극륭중적활성수용성음성대조조위12.019±0.830화5.388±0.480,단삼주사액조위4.445±1.061화2.856±0.597,F=31.78,P＜0.01；지용성음성대조조위14.155±0.672화4.299±0.252,단삼동ⅡA조위9.638±0.854화3.192±0.450,F=24.10,P＜0.01.결론 단삼가억제계통성경화병고효원합성성섬유세포극륭Ⅰ형전효원기인적전록,기단체단삼동ⅡA화원인다철가능발휘주요작용.
Objective To study the transcriptional regulation of type Ⅰ procollagen gene in systemic scleroderma(SS)-derived high collagen-producing fibroblast clones by Radix Salviae Miltiorrhizae(RSM).Methods Fibroblast clones with different collagen-producing capacity were previously obtained from patients with SS and normal human controls,and divided into 5 groups to be treated with RSM(1 g/L)injection,its water-soluble active monomers including sodium danshensu(20 mg/L),salvianolic acid B(5 mg/L)and protocatechuic aldehyde(5 mg/L),and lipid-soluble active monomer(tanshinone Ⅱ A,5mg/L)respectively.The fibroblast clones incubated with no drugs served as the water soluble negative control group,and those with dimethyl sulfoxide(DMSO)as the lipid soluble negative control group.MTT assay was performed to evaluate the proliferation of the fibroblast clones after 1-,3-,5-,and 7-day treatment,transient transfection and dualluciferase reporter assay system to quantify the relative activity of collagen type Ⅰ,alpha 1(COL1A1)proximal promoter in these fibroblast clones.Results The inhibitory effect of RSM and its active monomers on the proliferation of fibroblast clones was inapparent within the initial 3 days(P ＞ 0.05),but was enhanced with incubation time.A significant difference was observed in the proliferation level of fibroblast clones between RSM group and water-soluble negative control group on day 5(q′ =3.22,P ＜ 0.01),between RSM,salvianolic acid B,protocatechuic aldehyde groups and the water-soluble negative control group(q′ =4.74,3.03,2.56,all P ＜0.05)on day 7,and between tanshinone Ⅱ A and lipid-soluble negative control group on day 5 and 7(t =2.22,2.15,both P ＜ 0.05).RSM injection,tanshinone Ⅱ A and protocatechuic aldehyde significantly inhibited COL1A1 proximal promoter activity in SS-derived and normal control fibroblast clones(all P ＜ 0.01),and the former two drugs preferentially downregulated COL1A1 proximal promoter activity in SS-derived high collagenproducing fibroblast clones.Significantly different COL1A1 proximal promoter activity was observed in SS-derived high and low collagen-producing fibroblast clones between water-soluble negative control group and RSM injection group(12.019 ± 0.830 vs.4.445 ± 1.061,5.388 ± 0.480 vs.2.856 ± 0.597,F=31.78,P＜ 0.01),and between lipid-soluable negative control group and tanshinone Ⅱ A group(14.155 ± 0.672 vs.9.638 ±0.854,4.299 ± 0.252 vs.3.192 ± 0.450,F=24.10,P＜ 0.01).Conclusions RSM inhibits the transcription of COL1A1 gene in SS-derived high collagen-producing fibroblast clones,which may be mainly attributed to tanshinone Ⅱ A and protocatechuic aldehyde.
