摘要
一、研究目的
立足系统性红斑狼疮(SLE)免疫失衡角度,从临床研究和动物实验,观察Thl／Th2细胞群偏移及血管损伤因子与SLE发病机制关系,探讨SLE发病机理；分析清养透解法对该平衡的调节和干预血管损伤的作用,探讨其治疗SLE的主要作用环节。
二、研究方法
(一)临床试验采用前瞻性单盲随机对照的方法,纳入观察病例共42例,其中治疗组22例,对照组20例,每组各2例脱失,另设12人为正常组。治疗组给予清养透解合剂(由青蒿10g、鳖甲15g、水牛角20g、生地15g、丹皮10g、玄参15g、秦艽10组成)及常规西药(以口服强的松片为主,病情复发或合并狼疮肾炎、狼疮脑炎等用口服激素难以控制病情时,加用标准环磷酰胺冲击疗法)治疗；对照组给予安慰剂及常规西药(同前)治疗,治疗6个月,观察治疗前后病情活动性评分、主要症状评分、中医证候积分变化以及评估治疗期间出现的药物副作用,取静脉血采用酶联免疫法进行Thl类细胞因子(IFN-γ、IL-12)、Th2类细胞因子(IL-10、IL-4)及血管损伤因子(sTM、vWF)检测。
(二)动物实验将24只8-10周雌性MRL/lpr小鼠按质量随机分为中药组、西药组、中西医结合组、模型对照组4组,每组6只,另取8-10周龄雌性BALB/C小鼠6只为正常对照组,中药组予清养透解合剂9.5g/kg生理盐水稀释按0.4ml/只,每日1次灌胃；西药组予强的松10mg/kg生理盐水稀释按0.4ml/只,每[211次灌胃；中西医结合组予强的松10mg/kg并清养透解合剂9.5g/kg生理盐水稀释0.4ml/只,每日1次灌胃；模型对照组予生理盐水按0.4ml/只,每日1次灌胃,连续给药6周,正常对照组则不给任何处理。6周后取血采用酶联免疫法进行Thl类细胞因子(IFN-γ、IL-12)、Th2类细胞因子(IL-10、IL-4)及血管损伤因子(sTM、vWF)检测,并取小鼠肾脏在光镜及透射电子显微镜下观察其病理变化。
三、结果
(一)临床实试验
1、一般临床资料：本研究纳入观察病例共42例,38人完成了试验,其中治疗组完成试验20例,对照组完成试验18例,另设12人为正常组,三组在性别、年龄组成比较无显著性差异(p>0.05),治疗组和对照组在脱失率、病程、病情分布上比较无显著性差异(p>0.05)。
2、疗效性临床资料分析：治疗组与对照组在治疗前病情活动性评分、主要症状评分、中医证候积分、强的松用量无显著性差异(p>0.05)；两组治疗前与治疗后6个月比较病情活动性评分、主要症状评分、中医证候积分、强的松用量有非常显著性差异,治疗6个月后明显低于治疗前(p<0.01)；治疗后6个月后两组间比较病情活动性评分、主要症状评分、中医证候积分、强的松用量有非常显著性差异,治疗组明显低于对照组(p<0.01)；2、对血清细胞因子的影响：治疗组和对照组治疗前与正常对照组比较血清Thl类细胞因子(IL-12、IFN-γ)、Th2类细胞因子(IL-10、IL-4)、Thl/Th2细胞因子比值(包括IFN-γ/IL-4、IL-12/IL-10的比值)、血管内皮损伤因子(包括sTM、vWF)均有非常显著性差异(p<0.01)；治疗6月与治疗前比较,治疗组能同时降低血清Thl类细胞因子(包括IL-12、IFN-γ)(p<0.05)、Th2类细胞因子(包括IL-10、IL-4)(p<0.05)、血管内皮损伤因子水平(包括sTM、vWF)(p<0.05),提高Thl/Th2细胞因子比值(包括IFN-γ/IL-4、IL-12/IL-10的比值)(p<0.05)；3、治疗6个月后出现的药物副作用：治疗组与对照组在柯兴氏征、高血压、消化性溃疡、脱发、继发白细胞减少、肝功能异常方面人数有非常显著性差异(p<0.01),治疗组发生这些副作用人数明显比对照组少。
(二)动物实验：1、血清Thl类细胞因子：中药组、西药组、中西结合组、模型对照组、正常对照组两两比较血清IL-12水平无显著性差异(p>0.05)；中药组、西药组、中西结合组、模型对照组间两两比较血清IFN-γ水平无显著性差异(p>0.05),但此四组小鼠与正常对照组比较血清IFN-γ水平均有显著性差异(p<0.05)；2、血清Th2类细胞因子：与正常对照组比较,模型对照组小鼠血清IL-10、IL-4水平升高,有显著性差异(p<0.05)；与模型对照组小鼠,中药组、中西医结合组能同时降低血清Th2类细胞因子(包括IL-10、IL-4)水平,西药组可降低血清IL-4水平,有显著性差异(p<0.05)；3、对Thl/Th2平衡的影响：中药组、西药组、中西医结合组、模型对照组与正常对照组比较,IL-12/IL-10及IFN-γ/IL-4比值水平下降,有显著性差异(p<0.05)；与模型对照组比较,中药组、西药组、中西医结合组三组能提高MRL/lpr血清IFN-γ/IL-4比值(p<0.05),中西医结合组还可提高血清IL 12/IL 10的比值,有显著性差异(p<0.05)；4、对血管损伤因子的影响：模型对照组与正常对照组比较,血清vWF、sTM水平均上升,有非常显著性差异(p<0.01)；与模型对照组比较,中药组、中西结合能同时降低血清sTM、vWF水平,西药组能降低血清vWF水平,有显著性差异(p<0.05)；5、对肾脏病理学影响：MRL/lpr小鼠肾脏的光镜下检查可见肾小球增生或萎缩,大小不均匀,肾小球系膜区增宽,基底膜节断性增厚,间质细胞增生,小血管管壁纤维素样坏死,血管周围大量慢性炎细胞浸润；部分肾小管上皮细胞变性、坏死、脱落；电镜下可见肾小球基底膜增厚,足突细胞部分融合,内皮细胞肿胀,系膜基质较多,内皮下、基底膜、系膜区可见电子致密物。中药组、西药组、中西结合组能减轻肾脏病理改变。
四、结论
(一)临床试验提示,SLE患者体内存在Thl/Th2细胞平衡紊乱,Thl/Th2细胞向Th2漂移,血清vWF、sTM表达增高。以清养透解合剂为主的中西医结合治疗组在减轻SLE病情活动、改善SLE主要症状及阴虚证候方面明显优于单纯西药对照组；中西医结合治疗能改善Thl/Th2细胞群失衡,降低血清vWF、sTM的表达,减轻血管损伤；加速强的松的撤减,减少强的松用量,减轻强的松及环磷酰胺引起的毒副作用。
(二)动物实验提示,MRL/lpr小鼠存在Th1/Th2细胞平衡紊乱,主要表现为Th2升高,Th1/Th2细胞向Th2漂移,血清vWF、sTM的表达增高,肾脏组织发生病变,清养透解合剂能降低Th2的表达,提高IFN-γ/IL-4比值,降低血清vWF、sTM的表达,改善肾脏病变。
(三)动物实验与临床试验存在差异,MRL/Lpr小鼠血清Thl类细胞因子与正常对照组比较,表达降低或差异无显著性意义,而SLE患者血清Th1类细胞因子水平均明显增高。人和动物研究结果之所以存在差异性,可能是由于MRL/lpr小鼠与SLE患者发病机制不完全一致所致。
(四)清养透解法可能通过改善Th1/Th2细胞失衡,从而维持T细胞功能正常,抑制B细胞的异常增殖,减轻因炎症因子的分泌和免疫复合物在组织沉积形成的炎性损伤,保护血管内皮而达到治疗SLE作用的。
Objectives:Focusing on the imbalance of immunity in SLE, the study was to investigate the imbalance of Th1 and Th2 cell immunity as well as vascular damage factors in the pathological process of SLE. Clinical experiments were carried out to explore the effect of Qingyangtou jie therapy used to treat SLE.
Methods:
1、Clinical trial:Prospective a single blind Randomized Controlled trial was adopted, and 42 cases were divided into treatment group (22 cases) and control group (20 cases). We selected 12 persons as a normal group in Addition. The patients in treatment group were treated with a therapy with Qing-yang-tou-jie mixture and western medicine, while those in control group were treated with placebo and western medicine. The patients' condition activity scores, main symptom scores, TCM syndrome scores and side effects of cortin were evaluated after 6 month of treatment. TH1 cell factors (IFN-γ, IL-12), TH2 cell factors (IL-10, IL-4) and vascular injury factors (sTM, vWF) in serum were tested by ELISA simultaneously.
2、Animal experiment:Twenty-four female MRL/lpr mice of 8 to10 weeks were divided into therapentic group, western medicine treatment group, combined Chinese and western medicine treatment group and model control group. Six female BALB/C mice of 8 to 10 weeks were selected as a normal group. After continuous exposure therapy of six weeks, TH1 cell factors (IFN-γ, IL-12), TH2 cell factors (IL-10, IL-4) and vascular injury factors (sTM, vWF) in serum were tested; renal pathological changes and ultrastructure were observed by light microscopy and electronic microscopy.
Result
1、Clinical trial:
Thirty-eight cases in the clinical trial were recorded completely, which involves 20 treatment cases,18 control cases and 12 normal control cases. There was no difference in sex and age of these cases. There was no difference in the case lost rate, disease progress between therapentic and control groups. (1)Clinical effects:After 6 month of treatment, the patients'condition activity score, main symptom score, TCM syndrome score and diversity of prednisone dosage were significantly ameliorated than those of pre-treatment. The improvement in the condition of treatment group was significantly superior to the control group (P<0.01).
(2) Influence on cell factors:Before treatment, compared with normal group, the factors such as IL-12, IFN-γ, IL-10 and IL-4 in SLE patients increased obviously (p<0.05), and IL-12/IL-10, IFN-γ/IL-4 decreased significantly (P <0.01). Vascular injury factors also decreased significantly. After 6 month of treatment, the serous cell factors (both Thl and Th2 cell factors) decreased significantly in the treatment group, and vascular injury factor (sTM, vWF) decreased too. At the same time, Th1/Th2 ratio (both IFN-γ/IL-4 and IL-12/IL-10) in the treatment group increased, the difference was significant.
(3) Compared with control group, the treatment group had reduced incidences of side effects, such as Cushing syndrome, hypertension, peptic ulcer, alopecia, secondary leucopenia and liver dysfunction, and the differences were extraordinary remarkable (P<0.01)
2、Animal experiment:
(1)Th1 cell factors:No significant differences in IL-12 level were found in these five groups, independently (p>0.05), but there was significant difference in IFN-γbetween MRL/lpr mice and BALB/C mice (p<0.05);
(2)Th2 cell factors:Compared with normal group, the model mice's serous level of IL-10 and IL-4 increased significantly (p<0.05). While the level of Th2 cell factors (IL-10, IL-4) decreased significantly in the therapentic group and the combined Chinese and western medicine treatment group. The serous level of IL-4 decreased in western medicine group.
(3)Th1/Th2 balance:The ratio of Th1/Th2 (both IL-12/IL-10 and IFN-γ/IL-4) decreased in all groups except the normal group. But the ratio of IFN-γ/IL-4 was increased significantly in the therapic group, the western medicine group and the with group combined Chinese and western medicine. The serous level of IL-12/IL-10 was effectively increased in Chinese and western medicine group (p<0.05);
(4)Vascular injury factor:The serous vWF and sTM increased greatly in model mice (MRL/lpr mice). The therapentic treatment and the combined Chinese and western medicine treatment lowed greatly both vWF and sTM in the model mice. Western medicine treatment lowered the level of serous vWF in model mice.
(5)Renal Pathology:Under microscope, glomerulus proliferation or atrophy were found by observing renal pathological sections. There were also capillo fibrinoid necrosis and chronic inflammatory cells infiltrated around blood vessels. Under electron microscope, incrassation of basilar membrane, swollen endothelial cells, and dense deposits near basilar membrane were observed. All the treatments might relieve the micro pathology to some extent.
Conclusion
1. According to the result of clinical experiment, the balance of Th1/Th2 was disturbed in SLE patients, with the rise of Th2 and increase in the expression of vWF and STM in the serum. Qing-yang-tou-jie as the principal treatment in combination with western medicine was effective to reduce symptoms of SLE and Yin-deficiency, and the effect was superior to that of west medicine alone. The combined therapy was effective to improve the balance between Th1/Th2 cells, reduce the expression of vWF and sTM in the surum, relieve the vascular injury, and reduce the dosage of prednisone as well as its toxic side effects.
2. Experiment on animal showed that MRL/lpr mice suffered disturbance in Th1/Th2 balance, which was reflected by increase of Th2, decrease in Th1/Th2, increased expression of vWF and sTM in the serum, and pathologic changes in the kidney tissue. Qing-yang-tou-jie therapy was able to decrease the expression of Th2, increase IFN-γ/IL-4 ratio, suppress the expression of vWF and sTM in the serum and relieve pathological changes of kidney.
3. There was difference in the results of the animal experiment and clinical experiments. The Th1 cell factors in MRL/Lpr mice were lower or had no difference compared with the normal control mice, while in SLE patients, serum Th1 factors were significantly higher than in the control group. The difference in the results between the model mice and SLE patients might be due to difference in disease mechanisms.
4 Qing-yang-tou-jie therapy effectively to ameliorate the imbalance of Th1/Th2 cell factors, and thus helped to maintain the normal function of T cells, suppress the abnormal reproduction of B cells, reduce the inflammatory injury caused by the deposition of inflammatory factors and immune complex, and protect vascular endothelium, achieving the effect to cure SLE.
引文
[1]刘喜德.论肾虚瘀毒为系统性红斑狼疮的基本病因病机[J].中国中医药信息杂志[J],2002,9(12)：4-5.
[2]于佐文,金实.金实教授系统红斑狼疮证治经验探讨[M].硕士毕业论文：14
[3]许伟英,任世禾.中医药治疗系统性红斑狼疮机理探讨[J].光明中医,2007,22(5)：36-37。
[4]榻国维,范瑞强,陈达灿.中医皮肤病临证精粹[M].广州：广东人民出版社,2001,150-1561。
[5]王萍,张广中,王禾,等.养阴益气解毒法治疗系统性红斑狼疮探讨[J].中国中西医结合皮肤性病学杂志,2003,2(2)：79-82.
[6]周仲瑛.“伏毒”新识[J].世界中医药,2007,2(2)：73-75.
[7]赵智强,周仲瑛.从阳毒辨治红斑狼疮[J].中医药学报：1998,4：18-19。
[8]眭书魁,马秀清,董燕平,等.系统性红斑狼疮的中医病机研究[J].河北中医：2002,24(2)：153-154。
[9]许德清,曾凡钦.红斑狼疮[M].中国医药科技出版社.2003,6(1)：79.
[10]Morimoto S et al. The imcreased interieukin-13 in patients with systemic lupus eryrhematosus; relations to other Th1、Th2 related cytokines and clinical findings[J]. Auto immunity 2001,34(1):19-25.
[11]王少敏,张洁,马丽,等.30例女性SLE患者淋巴细胞培养上清液中IL-2、TNF-α、IFN-β、IL-4水平测定[J].中国妇幼保健,2008,23(7)：982-983.
[12]MajoJ, GhezzoH, CosioMG. [J].Eur RespirJ,2001,17:946-953.
[13]吴元胜,欧阳杰,榻国维,等.狼疮Ⅱ号对缺乏淋巴增殖基因狼疮鼠Thl／Th2细胞因子谱的影响[J].广州中医药大学学报,2009,26(4)：363-427.
[14]谢红付,李捷,施为,等.皮质类固醇激素对系统性红斑狼疮患者Th细胞因子平衡的影响[J].湖南医科大学学报,2002,27(6)：533-535.
[15]弓娟琴,陈志强,胡兹嘉,等.系统性红斑狼疮患者外周血单个核细胞中IL-4、IL-13 mRNA的表达[J].中国免疫学杂志.2001,17：500.
[16]Viallard JF.Th1(IL-2, IFN-γ) and Th2(IL-4, IL-10)cytokine production by peripheral blood mononuclear cells(PBMC) from patients with systemic lupus erythem tosus(SLE)[J]. Clin Exp Immunol,1999,115(1):189-195.
[17]石韫珍,孟锐,李丽,等.青藤碱对红斑狼疮患者外周血细胞的抑制作用及机制探讨[J].中国免疫学杂志,2008,24(9)：800-807
[18]吴昌辉,蔡川川,陆善词,等.系统性红斑狼疮患者Th1、Th2细胞因子 偏移与类风湿因子和抗心磷脂抗体的相关性研究[J].中国中西医结合皮肤性病学杂志,2005,4(3)：141-143.
[19]Reuss-Borst MA, Geyer A, Berner B. Cytokine-producing T cells in systemic lupus erythematosus:comment on the article by Akahoshi et al[J]. Arthritis Rheum,2000,43:711-712
[20]王晓栋,黎莉,沈南,等.Thl/Th2失衡在初发系统性红斑狼疮患者中的研究[J].中华风湿病学杂志,2002,6(5)：316-319.
[21]贺仆,孙涛.系统性红斑狼疮患者th1/th2失衡的研究[J].医学检验与临床,2008,19(3)：8-17.
[22]Tucci M, Lombardi L, Richards H B, Dammacco F, ext. Overexpression of interleukin-12 and T helper 1 predominance in lupus nephritis [J]. Clin Exp Immunol,2008,154:247-254.
[23]Horwitz DA, Gray JD, Behrendsen SC, et al. Decreased productionof interleukin-12 and other Thl-type cytokines in patients with recent onset systemic lupus erythematosus[J]. Arthritis Rheum,1998,41:838-844.
[24]章晓联主编.医学免疫学[M].武汉大学出版社,2008,1：61.
[25]Namjou B, Kilpatrick J, Harley JB.Genetics of clinical expression in SLE[J].Autoimmunity.2007 Dec,40(8):602-612.
[26]林梅青,宫奇林,孙涛,等.系统性红斑狼疮患者Thl、Th2细胞因子表达异常与T细胞亚群功能失调相关性研究[J].医学检验与临床,2007,18(5)：4-6.
[27]Uhm W S, Na K, Song G W, Jung S S, Lee T, Park M H, et al. Cy-tokine balance in kidney tissue from lupus nephritis patients [J]. Rheumatology (Oxford),2003,42:935-938.
[28]夏嘉,苏晓.干扰素γ与系统性红斑狼疮[J].第二军医大学学报,2010,31(9)：1020-1022.
[29]Chan R W, Lai F M, Li E K, Tam L S, Chow K M, Lai K B, et al. Intrarenal cytokine gene expression in lupus nephritis [J]. Ann Rheum Dis,2007,66:886-892.
[30]Hasegawa K, Hayashi T, Maeda K. Promotion of lupus in NZB XNZWF1 mice by plasmids encoding interferon(IFN) - gamma but not by those encoding interleukin (IL)-4 [J]. J Comp Pathol,2002,127:1-6.
[31]姜玉章,季晓辉,李玉峰,等.体外环磷酰胺和环孢素A对系统性红斑狼疮患者细胞因子释放及Th细胞亚群的作用研究[J].中华风湿病学杂志,2006,10(3)：161-165.
[32]李书芳.血栓调节蛋白在儿科领域的研究进展[J].中华现代临床杂志, 2005,3(21)：2233-2234.
[33]邹吉敏,袁宝军,苗丽娟,等.SLE患者血清IL-4、IL-10的检测及意义[J].中国煤炭工业医学杂志,2006,9(2)：178-179.
[34]陶洪群,李小龙,郑兴平,等.系统性红斑狼疮患者IL-18、IL-4和IFN-γ的变化及意义[J].医学研究杂志,2006,35(5)：48-49.
[35]Trinchieri GlInterleukin-12:a proinflammatory cytokine wit h im2munoregulatory fanctions t hat bridge innate resistance and antigen-specific adaptive immunity [J]. Annu Rev Immunol,1995,13:251-276.
[36]尚志华,于峥,杨建宇,等.浅述系统性红斑狼疮中药临床的几个问题.光明中医,2003,18(6)：52-54.
[37]邢美芬,刘晓华,姜玉章,等.系统性红斑狼疮26例IL-10及IL-12的异常表达及其相互作用[J].南京医科大学学报(自然科学版),2006,26(4)：283-287.
[38]Lauwerys BR, Van Snick J, Houssiau FA. Serum IL-12 in systemic lupus erythematosus:absence of p70 heterodimers but presence of p40 monomers correlating with disease activity[J]. Lupus,2002,11 (6):384-387.
[39]金彩云.调节性T细胞及细胞因子与系统性红斑狼疮[J].实用预防医学,2009,16(6)：1985-1987.
[40]Xue C, Lan-lan W, Bei C, et al, Abnormal Fas/ FasL and caspase-3-mediated apoptotic signalling pat hways of T lymphocyte subset in pa2tients wit h systemic lupus eryt hematosus [J]. Cell Immunol,2006,239 (2): 121-128.
[41]Wang H, Xu J, Ji X, et al. The abnormal apoptosis of T cell subsets and possible involvement of IL-10in systemic lupus eryt hematosus [J]. Cell Immunol,2005,235 (2):117-121.
[42]Crispin JC, Martinez A, Alcocer-Varela J. Quantification of regulatory T cells in patients with systemic lupus erythematosus[J].J Autoimmun, 2003,21:273-276.
[43]Kyriakos A., Kirouand Mary KC. New Pieces to the SLE cytokine puzzle[J]. Clin. Immunol.1999,91:1-5.
[44]巫向前主编.临床检验结果的评价[M],北京：人民卫生出版社[M],2000,4,1：219-223.
[45]张信强,刘晓华,柯瑶,等.系统性红斑狼疮患者血管内皮细胞损伤标志物检测及其临床意义[J].山东医药.2005,45(16)：11.
[46]Ahu ja A, Shupe J, Dunn R, et al. Dep letion of B cells in murine lupus: efficacy and resistance [J]. J Immunol,2007,179 (5):3351-3361.
[47]任文英,陈扬荣,郑全瑛,等.系统性红斑狼疮小鼠模型的研究近况[J].免疫学杂志,2003,19(3)：91-95.
[48]Atkinson C, Qiao F, Song H, et al. Low-dose targeted complement inhibition protects against renal disease and other manifestations of autoimmune disease in MRL/lpr mice[J]. J Immunol,2008,180 (2):1231-1238.
[49]Izui S, Merino R, FossatiL, et al. The role of the Yaa gene in lu2 pus syndrome[J]. Int Rev Immunol,1994,11 (3):211-230.
[50]余立凯,黄安斌,侯晓华,等.自发性狼疮小鼠模型研究进展[J].国际病理科学与临床杂志,2009,29(2)：82-85.
[51]朱方石,金实,汪悦,等.狼疮静颗粒对自发性狼疮小鼠肾狼疮抑制作用的实验研究[J].中国中西医结合杂志,2004,24(4)：343-347.
[52]钟嘉熙,彭胜权,张剑勇,等.中西医结合治疗系统性红斑狼疮25例[J],中国中西医结合杂志1999,19(1)：47-48.
[53]张剑勇,钟嘉熙.苓丹片、青蒿琥酯对系统性红斑狼疮患者IL-2与sIL-2R影响的研究[J].河南中医学院学报,2003；18(2)：38-39.
[54]张剑勇,钟嘉熙,戴馨仪,等.苓丹片、青蒿琥酯对SLE小鼠T淋巴细胞亚群的影响[J].中药药理与临床,1998；14(6)：27-29.
[55]张剑勇,钟嘉熙,史志云,等.苓丹片与青蒿琥醋对系统性红斑狼疮患者T细胞亚群的影响[J].中国中西医结合杂志,2002,22(7)：489.
[56]陈银环,钟嘉熙,刘叶,等.清养透解法为主综合治疗系统性红斑狼疮患者例生存质量评价[J].新中医,2009,41(1)：47-48.
[57]高学敏,许占民,李钟文.中药学[M],北京：人民卫生出版社,2000,1：580,543
[58]叶燕霞,曾耀英,黄秀艳,等.双氢青蒿素对小鼠T细胞的免疫抑制作用[J].中国病理生理杂志,2010.,26(3)：417-42.
[59]米绪华,栾森,马行,等.青蒿琥酯对IgA肾病模型大鼠肾组织MCP—1及其mRNA表达的影响[J].四川大学学报(医学版)2009,40(5)：821-825.
[60]王红,姜波,张华勇,等.青蒿琥酯通过抑制ICAM—1治疗鼠狼疮性肾炎的研究[J].实用临床医药杂志,2010,14(17)：13.
[61]杜志峰,李振彬,孙春霞,等.青蒿琥酯对佐剂性关节炎大鼠血清TNF-α及IL-1β含量的影响[J].广东医学,2010,31(13)：1637-1640.
[62]唐尹萍,刘焱文.鳖甲研究概况[J].中国药师,2010,13(3)：423-425.
[63]王慧铭,孙炜.鳖甲多糖对小鼠免疫调节作用的研究[J].中国中药杂志,2007,32(12)：1245-1247.
[64]杨瑁,邹全明,王东听.鳖甲超微细粉免疫调节功能实验研究[J].食品科学,2000,21(3)：40-42.
[65]刘睿,段金廒,李友宾,等.水牛角主要药效学评价及解热活性物质基础研究[J].南京中医药大学学报,2007,23,(5)：297-301.
[66]李宝国,黄丽华,杜雪,等.水牛角不同相对分子质量酶解液的药理作用研究[J].中国当代医药,2009,16(13)：16-17.
[67]拜玲,钟进芳,吴晓云,等.生地熟地中矿物元素的测定与分析[J].广东微量元素科学,2009,16(10)：50-55.
[68]王朴.生地黄的现代药理研究与临床应用[J].中国中医药现代远程教育,2008,6(8)：86.
[69]赵润生,张一听,苗冬雪,等.生地黄对血瘀模型大鼠血液流变性的影响[J].中药药理与临床,2006,22(3)：123.
[70]谢小艳,夏春森.中药玄参的化学成分及药理研究进展[J].亚太传统医药,2010,6(5)：121-125.
[71]李静,陈长勋,高阳,等.玄参提取物抗炎与抗动脉硬化作用的探索[J].时珍国医国药,2010,21(3)：532-534.
[72]王春丽.牡丹皮化学成份及临床应用[J].中国实用医药,2010,5(25)：176-177.
[73]郭齐,李贻奎,王志国,等.丹皮酚药理研究进展[J].中医药信息,2009,26(1)：20-22.
[74]徐泽红.中药秦艽的研究进展[J].中国医药导报,2008,5(6)：29-30.
[75]高兰月,高亦珑,赵淑红.秦艽对心血管的作用及毒性研究[J].中国民族民间医药,2010,19(11)：24.
[76]李永平,李向阳,王树林,等.秦艽提取物抗病毒的药效学实验研究[J].时珍国医国药,2010,21(9)：2267-2269.
[77]系统性红斑狼疮诊断标准[J].疑难病杂志,2007,6(4)：227.
[78]Bombardier C, Gladman DD,Vrowitz MB, et al. Derivation of the SLEDAI:a disease activity index for lupus patients. Arthritis Rheum,1992,35:630-640.
[79]郑筱萸主编.中药新药临床研究指导原则[M],北京：中国医药科技出版社,2002,1：111115.
[80]刘喜德.论肾虚瘀毒为系统性红斑狼疮的基本病因病机[J].中国中医药信息杂志[J],2002,9(12)：4-5.
[81]张曲矗,陆进明.系统性红斑狼疮与细胞因子的研究进展[J].现代生物医学进展[J],2009,9(19)：3794-3797.
[82]邵松,刘义庆,许丽,等.性激素对狼疮模型鼠Th1/Th2类细胞因子分泌的影响[J].现代检验医学杂志,2009,24(3)：31-33.
[83]杨会君,徐海丰,王亚斐,等.曲古抑菌素A对SLE患者外周血IL-10和IL-12分泌的影响[J].中国麻风皮肤病杂志,2006,9：20-22.
[84]孔海云主编.现代自身免疫病学[M],北京：人民军医出版社,1996,1：151-183.
[85]喻文球.论系统性红斑狼疮之阴精亏损[M],北京：世界科学出版社,1999,1：164.
[86]苏晓.沈王安教授治疗系统性红斑狼疮的经验[J].新中医,1998,30(8)：10.
[87]安家丰.张志礼.皮肤病医案选萃[M],北京：人民卫生出版社,1994,1：61.
[88]陈银环,钟嘉熙.钟嘉熙运用伏气温病理论辫治杂病经验[J].辽宁中医学欢学报,2005,11(6)：565-566.
[89]Velasquez X, Verde jo U, Massardo L, Martinez ME, Arriagada S, Rosenberg H, Valdivieso A, Jaeobelli S. Outeome of Chilean Patients with Lupus Nephritis Response to Intravenous CyeloPhosphamide. J Clin Rheulnatol.2003;9(1):7-14。
[90]董熠,刘秀珍,刘从彬,等.血栓调节蛋白的临床研究进展[J].安徽医药,2010,14(4)：383-384.