Experimental Study On Effects Of Wen YangQuYu Method On Scleroderma Skin Fibroblasts

Objective To investigate the pharmacologic mechanism of WenYangQU Yu(WYQY)method in the treatment of sysmetic sleroderma(SSc).Methods (1)Making medicated serum: According to the difference of therapeutics, twelve patiens were divided into four groups: the control group(3 cases),the medicined group(3 cases),the traditional Chinese medicine(TCM) group(2cases).the traditional Chinese and western medicine group (TCM-WM) group(4case). (2)Normal dermal fibroblasts and scleroderma lesional dermal fibroblasts were established from skin biopsies of lesional areas of the patiens with scleroderma and healthy human skin by tissue culturing method in vitro.(3) The proliferation of scleroderma and normal skin firoblasts was measured by MTT assay. (4)Flow cytometry were performed to observe the effects of medicated serum of WYQY herbs on scleroderma dermal fibroblastic cycle.(5) we investigated collagen production by Hydroxyproline Colorinmetric analysis.Results (1)Medicated serum of the treatment groups could change the morphology of fibroblasts derived from scleroderma and normal skin, respectively. (2)The medicated serum of different concentrations significally inhibited normal skin fibroblasts proliferation compared the control group (P< 0.01),and the TCM-WM groupe more inhibited than the western medicine group(P< 0.01).At the same time , the medicated serum of different concentration of the western medicine group and the TCM-WM group significally inhibitedscleroderma skin fibroblast proliferation compared with the control group(P < 0.01),but 20% only the medicated serum the TCM group inhibited its proliferation(P < 0.01); 20% the TCM-WM group more inhibited its proliferation than the western medicine group(P < 0.01).@20% the medicated serum of the TCM groups significally increased the percentage of G0-G1 phase(P < 0.01) and reduced the percentages of S and G2-M phase in the normal fibroblastic cycle (P < 0.01).At the same time,20%the medicated serum of the TCM groups significally increased the percentage of Go-Gi phase and reduced the percentages of S and G2-M phase in the sleroderma dermal fibroblastic cycle (P < 0.01).Cultrued scleroderma skin fibroblasts significally increaseed collagen synthesis compared with normal fibroblasts in vitro(p<0.01).Conclusions (1)WYQY method significally inhabite the growth of SSc skin fibroblasts and the integrated traditional Chinese and western medicine is superior to western medicine or TCM alone in inhibition of scleroderma skin fibroblasts,indicating of synergistic effect TCM herb and the western medicine. (2)WYQY method decrease overproduction of ECM totally through inhibiting scleroderma lesional fibroblast proliferation , this is one possible mechanism in treatment ofSSc. cycleis probabelly is one of mechanism of inhibition of scleroderma skin fibroblasts proliferation. (4)WYQY method unselectively interfere with normal and scleroderma skin fibroblasts.(5)Collagen synthesis is elevated in the scleroderma skin fibroblasts in vitro.It is one of pathlogy of SSc.