Effect Of Yiqiruanpi Decoction Ultramicro-drug Piece On Systemic Sclerosis By Clinical And Exper-imental Research

CHAPTER I:A retrospective study on 236 cases of systemic sclerosis treated with Yiqiruanpi PillObjective:To explore the pathogenic factors, clinical features, and the correlative factors influencing the effectiveness of SSc,and to evaluate the clinical effectiveness and safety of Yiqiruanpi Pill on treating systemic sclerosis(SSc).Method:Clinical data of 236 cases of SSc treated with Yiqiruanpi Pill were retrospectively analyzed, including sex, age, course of disease and treatment,initial symptoms,family history, clinical manifestations, laboratory examinations, untoward effect and therapeutic effect.All available data were dealt with statistical analysis and correlative conclusions were made.Results:1. Of the 236 patients 81 were males(34.3%)and 155 were females(65.7%), the ratio of males to females was 1 to 1.91.The mean age of onset was 41.08±11.73 years old and the male's was 44.91±10.87 years old and the female's was 39.08±12.22 years old (P<0.01). The mean course of disease was 3.71±4.11 years and the male's was 3.15±4.05 years and the female's was 4.01±4.13 years (P<0.01).168 cases had Raynaud's phenomenon as their initial symptom (71.2%) and 4 cases had family history.2. The total effective rate was 95.3%.The principal symptoms and physical signs and partial laboratory examinations were significantly improved.The effectiveness of Yiqiruanpi Pill was optimal to swollen phase and secondary to scleritic phase and inferior to atrophy phase. The course of treatment had distinct effect to the effectiveness of Yiqiruanpi Pill and over 9 months' treatment had better effectiveness.3. There's a higher safety of Yiqiruanpi Pill on treating SSc.Conclusion:Yiqiruanpi Pill is effective for SSc and has a higher safety.CHAPTERⅡ:Clinical research on 32 cases of SSc treated with Yiqiruanpi Decoction ultramicro-drug.Objective:To evaluate the clinical effectiveness of Yiqiruanpi Decoction ultramicro-drug(YDUD) on treating SSc,and to explore its effect on circulating endothelial cell(CEC) and endothelin-1(ET-1).Method:32 patients with SSc were divided randomly into two groups, treated with traditional Yiqiruanpi Decoction(YD) and 1/3-dose YDUD Piece respectively in 3 months.The changes of skin sclerosis scores, minimal middle finger-to-palm distance,maximal upper-to-lower teeth distance, the numbers of CEC and the plasma levels of ET-1 were measured respectively before and after treatment,and the therapeutic effects were evaluated.Results:1. The total effect rate of YD group was 87.5% and the 1/3YDUD group was 93.8%. The clinical effectiveness of the two groups were equal (p>0.05)2. Comparison of pre-treatment, YD group and 1/3YDUD group could both remarkably reduce skin sclerosis scores and minimal middle finger-to-palm distance and improve maximal upper-to-lower teeth distance (P<0.01). And there were no significant difference between them (p>0.05)3. Comparison of pre-treatment, YD group and 1/3YDUD group could both significantly reduce the number of CEC and down-regulate the plasma level of ET-1 (P<0.01). And there were no significant difference between them (p>0.05)Conclusions:1. YD can remarkably reduce skin sclerosis scores and minimal middle finger-to-palm distance and improve maximal upper-to-lower teeth distance, the efficiency between YD group and 1/3YDUD group are equal.2. YD can significantly reduce the number of CEC and down-regulate the plasma level of ET-1, and there were no significant difference between YD group and 1/3YDUD group.CHAPTERⅢ:Investigation of anti-fibrotic effect and its mechanisms of Yiqiruanpi Decoction ultramicro-drug on murine model of BLM- induced scleroderma.Objective:To investigate the anti-fibrotic effect of YDUD on murine model of BLM-induced scleroderma, and to explore the molecular mechanisms of this effect.Method:1.72 SPF female BALB/c mice were randomly divided into 6 groups (12 in each group):normal control group(A), model group (B), D-Penicillamine (D-Pen) group (C), YD group(D), YDUD group (E) and 1/3-dose YDUD group (F). Group B, C, D,E and F received daily subcutaneous injection of 0.1 ml of BLM (300μg/ml, diluted in saline), and group A received 0.1ml of saline;Meanwhile,group C, D, E and F were treated with 125mg/kg D-Pen,16.6g /kg YD,16.6g/kg YDUD,5.5 g/kg YDUD, and group A and B were treated with the same volume of saline, respectively,all by daily i.g. for 4 weeks.2.On the next day after the final treatment the back skin pieces of all these mice were removed for histological examination, measurement of dermal thickness,determination of hydroxyproline contents by colorimetric method, determination of CTGF mRNA expression by in situ hybridization (ISH) and COL-Ⅰ,Ⅲ, TGF-β1 by immunohistochemistry. The right-side lungs were also excised for histological examination.Results:1.The model mice skin and lungs showed significant sclerosis histologically compared with normal control group,and all treatment groups had less sclerosis than the model group.2.The dermis of model group was significantly thicker than that of normal control group and all treatment groups (P<0.01).The skin hydroxyproline contents of model group were significantly higher than those of normal control group and all treatment groups (P<0.01).There were no marked differences in group E and group F compared with group D(P>0.05).3. The integral optical density(IOD) of the skin CTGFmRNA of model group was significantly higher than that of normal control group and all treatment groups (P<0.01). There were no marked differences of IOD in group E and group F compared with group D(P>0.05).4. The integral optical density(IOD) of the skin TGF-β1 and COL-Ⅰ,Ⅲof model group was significantly higher than that of normal control group and all treatment groups (P<0.01). There were no marked differences of IOD in group E and group F compared with group D(P>0.05).5. The abundance of COL-Ⅰ,Ⅲof group D,E and F correlated closely with CTGFmRNA and TGF-β1.After treatment with Yiqiruanpi Decoction, the abundance of COL-Ⅰ,Ⅲwas reduced with the down- regulation of CTGFmRNA and TGF-β1.Conclusions:1.BLM can successfully induce scleroderma in BALB/c mice at a daily dose of 0.1ml (300μg/ml) by subcutaneous injection into the back for 4 weeks.2. YD can significantly prevent the development of dermal fibrosis induced by BLM.3. YD can down-regulate the expression of CTGF mRNA and TGF-β1, which may result in the down-regulation of the expression of COL-Ⅰand COL-Ⅲ.