养阴凉血方治疗过敏性紫癜性肾炎的理论及实验研究
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摘要
过敏性紫癜性肾炎是一种由免疫复合物(IC)介导的系统性小血管炎,本病的中心证候是血尿、蛋白尿、皮疹紫斑、关节肿痛,或见水肿等,属于祖国医学“血证”的范畴。其病机为风、湿、热毒之邪夹杂,热瘀阻络、络伤血溢,血溢脉外。早在清代,温病学派创始人叶天士就指出：“入血就恐耗血动血,直须凉血散血”,并倡导“务在先安未受邪之地”的治疗思想,即在疾病过程中要主动采取措施,防变于先,控制病势发展。本研究通过对“养阴凉血方”修复HSPN动物模型肾损害的研究,验证导师马健教授经验方“养阴凉血方”治疗过敏性紫癜性肾炎的免疫学、药效学机制,为其临床应用提供微观实验依据。
过敏性紫癜性肾炎为一种常见的血管变态反应性疾病,因机体对某些致敏物质发生变态反应,导致毛细血管脆性及通透性增加,血液外渗,产生皮肤、粘膜及某些器官出血；可同时伴发血管神经性水肿、荨麻疹等其他过敏表现；血管病变处有IgA和补体成分沉积为病理基础的临床综合征。多见于小儿,男女比例为2：1,肾脏累及在过敏性紫癜中十分常见,其严重程度并不与肾外表现相一致。观其临床表现,约30%～70%的患者出现血尿及(或)蛋白尿。据统计,过敏性紫癜性肾炎在所有小儿肾小球疾病中约占15%。本病病理共分VI型,光镜下肾穿刺发现本病与IgA肾病相似。典型的肾小球病变为系膜增生型肾小球肾炎伴不同程度的新月体形成。过敏性紫癜主要是免疫球蛋白A(IgA)介导的变态反应性疾病,免疫失控是其主要病理机制,体液免疫异常,IgA的作用尤其受到重视。T细胞功能紊乱和多种促炎症因子失调,并且其免疫复合物激活补体发挥作用。HSPN存在着免疫功能紊乱：Th2类细胞活化使IgA、IgE水平增高,尤其是IgA,形成IgA免疫复合物而致病,白细胞介素(IL)4、6、8及肿瘤坏死因子a(TNF a)作为调节细胞免疫重要细胞因子,在HSP／HSPN发病机制中起重要作用。长时间CD4/CD8比值降低是病情加重和预后不良的指标之一。外周血CD4+、CD4+/CD8+明显降低,而CD8+明显增高,表明T细胞亚群异常。CD4+减少,产生多种淋巴因子和协助B细胞产生抗体及辅助其它细胞亚群等功能减弱；CD8+增加,抑制CD4+和B细胞作用增强,影响细胞免疫反应和抗体形成。从而使机体免疫功能降低,导致感染反复发作和预后不良。
实验研究用双抗体夹心酶免疫吸附法(ELISA)测定血浆白细胞介素-1(IL-1)和TNF-α活性；采用免疫荧光检查肾脏(IgA、IgG、IgM);用流式细胞仪检测CD3+、CD4+、CD8+T细胞亚群等,通过免疫学检测,从免疫调节角度探讨了“养阴凉血方”方药防治过敏性紫癜性肾炎(HSPN)的机制。
本研究通过观察养阴凉血方配合激素治疗过敏性紫癜性肾炎的作用,初步结论如下：1、养阴凉血方配合激素对过敏性紫癜性肾炎有明显治疗作用,其可能机制：促进肝脏合成蛋白质增加,从而提高血清白蛋白水平,减轻血尿、蛋白尿,间接起到保护肾小球、肾小管间质的作用。同时抑制胆固醇合成,降低甘油三脂,并可改善肾脏微循环,减轻肾脏病变程度。2、养阴凉血方可能通过调整垂体-肾上腺系统,使降低的皮质醇水平恢复正常,减轻了激素的副作用,提高激素疗效。3、养阴凉血方能降低血浆IL-1及TNF-α含量,调整CD3+、CD4+及CD8+T细胞亚群的百分率或及其比值,表明该方对过敏性紫癜性肾炎大鼠的免疫功能可能有一定的调节作用。
Henoch-Schonlein purpura nephritis is an immune complex by the (IC)-mediated systemic small vessel vasculitis, the center of this disease syndrome are hematuria, proteinuria, rash, purpura, joint swelling, pain, or see edema and so on, belongs to Chinese medicine, "blood card (purpura, hematuria, blood in the stool)" category. The pathogenesis of wind, wet, hot toxic mixture of evil, network overflow injured blood, blood pulse overflow outside. As early as the Qing Dynasty, sent Febrile Diseases, founder of Ye Tianshi pointed out:"Blood on the fear of consumption of blood intothe movement of blood, straight to Liangxue casual blood," "service earlier an unaffected evil land" and the idea that in the disease process China should take the initiative to take measures to prevent changes in the first, control the development of illness. This study was the " Yang Yin Liang Xue Soup " Repair HSPN renal damagein animal model research, validation of experience mentor Professor Ma Jian, " YangYin Liang Xue Soup " treatment of allergic purpura nephritis immunity,pharmacodynamics mechanisms, their clinical application providing micro-experimental basis.
Henoch-Schonlein purpura nephritis is a common vascular allergic diseases, because the body's allergic reaction to certain allergens occurred, leading to increased capillary fragility and permeability, blood extravasation, resulting in the skin, mucous membranes and some organ hemorrhage; can simultaneously associated with angioedema, urticaria and other allergic manifestations; vascular lesions Department has IgA and complement component deposition of the pathological basis of the clinical syndrome. More common in children, male to female ratio of 2:1, and kidney in Henoch-Schonlein purpura involving the very common, its severity is not consistent with the extra-renal manifestations. View of its clinical manifestations, about 30%～70% of patients with hematuria and (or) proteinuria. According to statistics, Henoch-Schonlein purpura nephritis in all glomerular diseases account for about 15%. The disease pathology is divided into VI type, light microscopy, renal biopsy was found similar to this disease and IgA nephropathy. The typical glomerular lesions and mesangial proliferative glomerulonephritis with varying degrees of crescent formation. Henoch-Schonlein purpura is mainly immunoglobulin A (IgA)-mediated allergic diseases, the immune control is the main pathological mechanisms, humoral immune abnormalities, IgA, particularly the role of attention. T-cell dysfunction and a variety of pro-inflammatory factor disorders, and its immune complex activation of complement play a role. HSPN there is immune dysfunction: Th2 type cell activation so that IgA, IgE levels increased, especially IgA, the formation of IgA immune complexes and disease, interleukin (IL) 4,6,8 and tumor necrosis factor-α(TNFα) as an important cytokines regulating cell-mediated immunity, in the HSP/HSPN play an important role in the pathogenesis. So for the immune regulation function of the drug is present and future efforts.
Experimental research program to further in-depth study from revealing aspects of immune regulation medicine Febrile Diseases, "Yin cooling blood side" control sensitivity purpura nephritis (HSPN) mechanism. Double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) determination of plasma interleukin-1 (IL-1) and TNF-a activity; using immunofluorescence kidneys (IgA, IgG, IgM); by flow cytometry CD3+, CD4+, CD8+T cell subsets, by immunoassay, from the point of view of immune regulation, "nourishing yin and cooling blood side" Prescriptions for prevention and treatment of allergic purpura nephritis (HSPN) mechanism. Peripheral blood CD4+, CD4+/CD8+ significantly reduced, while CD8+increased significantly, indicating that abnormal T-cell subsets. CD4+ decrease, resulting in a variety of lymphokines and help B cells produce antibodies and complement other features such as reduced cell subsets; CD8+increase, inhibit the role of CD4+ and B cells enhanced affect cell immune response and antibody formation. Thereby reducing immune function, leading to recurrent infections and poor prognosis.
From the experimental study in-depth study reveals aspects of immune regulation Febrile Diseases in traditional Chinese medicine, " YangYin Liang Xue Soup " control sensitivity purpura nephritis (HSPN) mechanism. Double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) determination of plasma interleukin-1 (IL-1) and TNF-a activity; using immunofluorescence kidneys (IgA, IgG, IgM); by flow cytometry CD3+, CD4+, CD8+ T cell subsets, by immunoassay, from the point of view of immune regulation, " YangYin Liang Xue Soup "Prescriptions for prevention and treatment of allergic purpura nephritis (HSPN) mechanism.
This study by observing the " YangYin Liang Xue Soup "with hormone therapy Henoch-Schonlein purpura nephritis role, draw the following conclusions:1, " YangYin Liang Xue Soup "cooperate with the hormone on Henoch-Schonlein purpura nephritis significant therapeutic effects, its possible mechanisms:the promotion of Liver protein synthesis increase, thereby raising the level of serum albumin to reduce hematuria, proteinuria, indirectly, for protection of glomerular, tubular interstitial role. At the same time inhibit cholesterol synthesis, lower triglycerides and may improve renal microcirculation, reduce the degree of kidney disease.2, Yin Liangxue party may by adjusting the pituitary-adrenal system, so that lower cortisol levels return to normal, reducing the side effects of hormones, improve hormone effect.3, Yin cooling blood can only reduce the plasma IL-1 and TNF-a levels, adjust the CD3+, CD4+ and CD8+ T cell subsets and the percentage or ratio, indicating that the side Henoch-Schonlein purpura nephritis in rats immune features may have some regulatory role.
过敏性紫癜性肾炎为一种常见的血管变态反应性疾病,因机体对某些致敏物质发生变态反应,导致毛细血管脆性及通透性增加,血液外渗,产生皮肤、粘膜及某些器官出血；可同时伴发血管神经性水肿、荨麻疹等其他过敏表现；血管病变处有IgA和补体成分沉积为病理基础的临床综合征。多见于小儿,男女比例为2：1,肾脏累及在过敏性紫癜中十分常见,其严重程度并不与肾外表现相一致。观其临床表现,约30%～70%的患者出现血尿及(或)蛋白尿。据统计,过敏性紫癜性肾炎在所有小儿肾小球疾病中约占15%。本病病理共分VI型,光镜下肾穿刺发现本病与IgA肾病相似。典型的肾小球病变为系膜增生型肾小球肾炎伴不同程度的新月体形成。过敏性紫癜主要是免疫球蛋白A(IgA)介导的变态反应性疾病,免疫失控是其主要病理机制,体液免疫异常,IgA的作用尤其受到重视。T细胞功能紊乱和多种促炎症因子失调,并且其免疫复合物激活补体发挥作用。HSPN存在着免疫功能紊乱：Th2类细胞活化使IgA、IgE水平增高,尤其是IgA,形成IgA免疫复合物而致病,白细胞介素(IL)4、6、8及肿瘤坏死因子a(TNF a)作为调节细胞免疫重要细胞因子,在HSP／HSPN发病机制中起重要作用。长时间CD4/CD8比值降低是病情加重和预后不良的指标之一。外周血CD4+、CD4+/CD8+明显降低,而CD8+明显增高,表明T细胞亚群异常。CD4+减少,产生多种淋巴因子和协助B细胞产生抗体及辅助其它细胞亚群等功能减弱；CD8+增加,抑制CD4+和B细胞作用增强,影响细胞免疫反应和抗体形成。从而使机体免疫功能降低,导致感染反复发作和预后不良。
实验研究用双抗体夹心酶免疫吸附法(ELISA)测定血浆白细胞介素-1(IL-1)和TNF-α活性；采用免疫荧光检查肾脏(IgA、IgG、IgM);用流式细胞仪检测CD3+、CD4+、CD8+T细胞亚群等,通过免疫学检测,从免疫调节角度探讨了“养阴凉血方”方药防治过敏性紫癜性肾炎(HSPN)的机制。
本研究通过观察养阴凉血方配合激素治疗过敏性紫癜性肾炎的作用,初步结论如下：1、养阴凉血方配合激素对过敏性紫癜性肾炎有明显治疗作用,其可能机制：促进肝脏合成蛋白质增加,从而提高血清白蛋白水平,减轻血尿、蛋白尿,间接起到保护肾小球、肾小管间质的作用。同时抑制胆固醇合成,降低甘油三脂,并可改善肾脏微循环,减轻肾脏病变程度。2、养阴凉血方可能通过调整垂体-肾上腺系统,使降低的皮质醇水平恢复正常,减轻了激素的副作用,提高激素疗效。3、养阴凉血方能降低血浆IL-1及TNF-α含量,调整CD3+、CD4+及CD8+T细胞亚群的百分率或及其比值,表明该方对过敏性紫癜性肾炎大鼠的免疫功能可能有一定的调节作用。
Henoch-Schonlein purpura nephritis is an immune complex by the (IC)-mediated systemic small vessel vasculitis, the center of this disease syndrome are hematuria, proteinuria, rash, purpura, joint swelling, pain, or see edema and so on, belongs to Chinese medicine, "blood card (purpura, hematuria, blood in the stool)" category. The pathogenesis of wind, wet, hot toxic mixture of evil, network overflow injured blood, blood pulse overflow outside. As early as the Qing Dynasty, sent Febrile Diseases, founder of Ye Tianshi pointed out:"Blood on the fear of consumption of blood intothe movement of blood, straight to Liangxue casual blood," "service earlier an unaffected evil land" and the idea that in the disease process China should take the initiative to take measures to prevent changes in the first, control the development of illness. This study was the " Yang Yin Liang Xue Soup " Repair HSPN renal damagein animal model research, validation of experience mentor Professor Ma Jian, " YangYin Liang Xue Soup " treatment of allergic purpura nephritis immunity,pharmacodynamics mechanisms, their clinical application providing micro-experimental basis.
Henoch-Schonlein purpura nephritis is a common vascular allergic diseases, because the body's allergic reaction to certain allergens occurred, leading to increased capillary fragility and permeability, blood extravasation, resulting in the skin, mucous membranes and some organ hemorrhage; can simultaneously associated with angioedema, urticaria and other allergic manifestations; vascular lesions Department has IgA and complement component deposition of the pathological basis of the clinical syndrome. More common in children, male to female ratio of 2:1, and kidney in Henoch-Schonlein purpura involving the very common, its severity is not consistent with the extra-renal manifestations. View of its clinical manifestations, about 30%～70% of patients with hematuria and (or) proteinuria. According to statistics, Henoch-Schonlein purpura nephritis in all glomerular diseases account for about 15%. The disease pathology is divided into VI type, light microscopy, renal biopsy was found similar to this disease and IgA nephropathy. The typical glomerular lesions and mesangial proliferative glomerulonephritis with varying degrees of crescent formation. Henoch-Schonlein purpura is mainly immunoglobulin A (IgA)-mediated allergic diseases, the immune control is the main pathological mechanisms, humoral immune abnormalities, IgA, particularly the role of attention. T-cell dysfunction and a variety of pro-inflammatory factor disorders, and its immune complex activation of complement play a role. HSPN there is immune dysfunction: Th2 type cell activation so that IgA, IgE levels increased, especially IgA, the formation of IgA immune complexes and disease, interleukin (IL) 4,6,8 and tumor necrosis factor-α(TNFα) as an important cytokines regulating cell-mediated immunity, in the HSP/HSPN play an important role in the pathogenesis. So for the immune regulation function of the drug is present and future efforts.
Experimental research program to further in-depth study from revealing aspects of immune regulation medicine Febrile Diseases, "Yin cooling blood side" control sensitivity purpura nephritis (HSPN) mechanism. Double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) determination of plasma interleukin-1 (IL-1) and TNF-a activity; using immunofluorescence kidneys (IgA, IgG, IgM); by flow cytometry CD3+, CD4+, CD8+T cell subsets, by immunoassay, from the point of view of immune regulation, "nourishing yin and cooling blood side" Prescriptions for prevention and treatment of allergic purpura nephritis (HSPN) mechanism. Peripheral blood CD4+, CD4+/CD8+ significantly reduced, while CD8+increased significantly, indicating that abnormal T-cell subsets. CD4+ decrease, resulting in a variety of lymphokines and help B cells produce antibodies and complement other features such as reduced cell subsets; CD8+increase, inhibit the role of CD4+ and B cells enhanced affect cell immune response and antibody formation. Thereby reducing immune function, leading to recurrent infections and poor prognosis.
From the experimental study in-depth study reveals aspects of immune regulation Febrile Diseases in traditional Chinese medicine, " YangYin Liang Xue Soup " control sensitivity purpura nephritis (HSPN) mechanism. Double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) determination of plasma interleukin-1 (IL-1) and TNF-a activity; using immunofluorescence kidneys (IgA, IgG, IgM); by flow cytometry CD3+, CD4+, CD8+ T cell subsets, by immunoassay, from the point of view of immune regulation, " YangYin Liang Xue Soup "Prescriptions for prevention and treatment of allergic purpura nephritis (HSPN) mechanism.
This study by observing the " YangYin Liang Xue Soup "with hormone therapy Henoch-Schonlein purpura nephritis role, draw the following conclusions:1, " YangYin Liang Xue Soup "cooperate with the hormone on Henoch-Schonlein purpura nephritis significant therapeutic effects, its possible mechanisms:the promotion of Liver protein synthesis increase, thereby raising the level of serum albumin to reduce hematuria, proteinuria, indirectly, for protection of glomerular, tubular interstitial role. At the same time inhibit cholesterol synthesis, lower triglycerides and may improve renal microcirculation, reduce the degree of kidney disease.2, Yin Liangxue party may by adjusting the pituitary-adrenal system, so that lower cortisol levels return to normal, reducing the side effects of hormones, improve hormone effect.3, Yin cooling blood can only reduce the plasma IL-1 and TNF-a levels, adjust the CD3+, CD4+ and CD8+ T cell subsets and the percentage or ratio, indicating that the side Henoch-Schonlein purpura nephritis in rats immune features may have some regulatory role.
引文
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