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Research on Clinical Effects of Method of Heat-clearing, Detoxifying and Promoting Blood Flow (HCDP) on Lupus Nephritis (LN) and Its Mechanism

Author: LiYan
Tutor: HongQinGuo
School: Guangzhou University of Traditional Chinese Medicine
Course: Chinese medical science
Keywords: The method of heat-clearing, detoxifying the method of promoting blood flow Lupus Nephritis (LN) glomerular mesangial cell (GMC) apoptosis
CLC: R259
Type: PhD thesis
Year: 2007
Downloads: 248
Quote: 1
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Objective (1) To sum up the progress of the research on the pathogenic mechanismand prevention measures of Lupus Nephritis (LN) in traditional Chinese andWestern medical sciences. (2) To observe the clinical therapeutic effects onLN of toxic heat and blood stagnation with the method of heat-clearing,detoxifying and promoting blood flow (HCDP). (3) To observe the effects ofblood serum of type-ⅣLN patients on glomerular mesangial cell (GMC)proliferation of healthy people. To observe the effects of Chinese crude drugof HCDP on GMC proliferation and apoptosis, Bcl-2 protein synthesis andBcl-2mRNA expression. To explore the mechanism of the method of heat-clearing,detoxifying and promoting blood flow (HCDP) on the level of cell and molecularbiology and to provide experimental evidence for clinical application.Methods (1) Literature study: To consult a large number of ancient and modernresearch documents about Chinese and Western medical sciences, summarize andassess traditional Chinese medical science-oriented understanding,etiopathogenisis and Western medical science-oriented pathogenesis of LN, aswell as the status quo of researches into prevention and cure measures. (2)Clinical research: Sixty patients with LN of toxic heat and blood stagnationwere randomly and equally divided into a control group and a subject group.The control group adopted treatment of western medicine, while the subjectgroup was offered the method of heat-clearing, detoxifying and promoting bloodflow (HCDP) on top of western medicine. The period of treatment was three months.Cardinal symptoms included: erythema on face, febricity, aphtha, erythema onhands, arthralgia, arthrocele, myalgia, alopecia, puffiness, chest pain,palpitation, hypodynamia, dysphoria, feverish sensation in palms and soles, xerostomia, insomnia, tinnitus, ache, pharyngalgia and menstrual clots.Laboratory index contained: white blood cell (WBC), lymphocyte (LYMN),hemoglobin (HGB), platelet (PLT), urinary leukocyte (LEU), urinaryerythrocyte (ERY), urinary protein (PRO), blood urea nitrogen (BUN), serumcreatinine (Cr), plasma albumin (ALB), erythrocyte sedimentation rate (ESR),C3, C4, CH50, antinuclear antibody (ANA), anti-ds-DNA antibody, and bloodcoagulation function (including fibrinogen, FIB), etc. The Systemic LupusErythematosus Disease Activity Index (SLEDAI) and the integral syndrome ofTCM before and after the therapy were calculated. Therapeutic effect on thedisease and syndrome were respectively evaluated. (3) Empirical study:①Serum of healthy adults and serum of different concentration (5%, 10%, 15%and 20%) of type-ⅣLN patients were used to affect GMC, and their effectson GMC proliferation were observed. The serum (x%) whose effect on promotingGMC proliferation was the most significant was selected for the next experiment.②With the method of blood serum pharmacology, 5 groups were observed:group-Ⅰ: blank, group-Ⅱ: control serum+x% serum of type-ⅣLN patients,group-Ⅲ: low concentration serum which contained Chinese medicine of HCDP+x% serum of type-ⅣLN patients, group-Ⅳ: medium concentration serum whichcontained Chinese medicine of HCDP+x% serum of type-ⅣLN patients, group-Ⅴ: high concentration serum which contained Chinese medicine of HCDP+x%serum of type-ⅣLN patients. MTT method was adopted to detect theproliferation and inhibition ratios of GMC. Apoptosis of GMC were detectedwith the fluorometric method and flow cytometer respectively.③Afterwards,the methods of western blot and RT-PCR were respectively used to detectsynthesis of Bcl-2 protein and expression of Bcl-2mRNA , which was the genein GMC inhibiting apoptosis, by 5 groups (group-1: blank, group-2: controlserum+x% serum of type-ⅣLN patients, group-3、4、5: low、medium and highconcentration serum which contained Chinese medicine of HCDP+x% serum oftype-ⅣLN patients respectively).Results (1)Study on publications:①Name: According to clinical manifestations,LN was discovered in diseases described in traditional Chinese medicalsciences, such as "toxin of Yin-Yang", "red butterfly-shaped sore","patch of Yang toxin", "arthromyodynia", "edema", "kidney deficiency"and so on.②Etiopathogenisis and pathogenesis: The etiopathogenisis andpathogenesis of LN of toxic heat and blood stagnation was summarized as Yin asthenia of liver and kidney, toxic heat and blood stagnation.③Therapy: Themain TCM therapeutic methods of LN include: treatment by types, by stages,by Chinese patent medicine, by private prescription, by a single medicinalherb, by extract from or injection of traditional Chinese medicine, etc. Thetherapeutic effects on LN are satisfactory. Nowadays, most academicians adoptthe method of HCDP to cure LN.④The study of modern medical science: It isconsidered that the incidence of LN is very complicated. It is in connectionwith genetic factors (genetic predisposing factors, etc), abnormal GMCproliferation and apoptosis, dysfunction of immunoregulation (imbalance ofcytokine network, declining ability to generate autoantibody like anti-ds-DNAantibody and anticardiolipin antibody and clean immune complex), as well asneural, endocrine and metabolic factors, etc. The abnormal generation andapoptosis of intrinsic glomerular cells such as GMC is an important mechanismof the incidence and evolvement of LN. Therefore, prevention and cure methodsare to inhibit proliferation and induce apoptosis of intrinsic glomerularcells like GMC. Therapeutic methods contain glucocorticoid, immunosuppressiveagent, other treatments such as plasmapheresis, immunoadsorption (IA),immunoglobulin, etc. But Western medical science-oriented therapy has manydisadvantages, such as non-standard therapeutic programs, insensitivity ordrug tolerance of some patients, considerable side effects, and so on.Therefore, researches focus on studying TCM therapeutic methods of tangibleeffects, economy and few side effects. (2)Clinical research: In the subjectgroup, the total effective rates of the disease and syndrome of TCM were 83.33%and 93.33% respectively. The effective rate of the disease was higher thanthe control group, but the difference had no statistical significance (P>0.05). The effective rate of the syndrome was also higher compared with thecontrol group and the discrepancy was extremely significant (P<0.01). In thesubject group, SLEDAI and the integral syndrome of TCM after therapy wereremarkably lower than before, and the discrepancies were extremely significant(P<0.01). After therapy, the reduction of SLEDAI in the subject group wasmore satisfactory compared with the control group but the difference was notsignificant (P>0.05); while the integral syndrome of TCM was remarkably lowerthan the control group, and the difference was extremely significant (P<0.01). After therapy, main symptoms of the patients in the two groups wererelieved notably compared with symptoms before therapy and the differences were extremely significant (P<0.01). In the subject group, the effect ofmoderating main symptoms of LN was better than the control group, but thediscrepancy was not significant (P>0.05). Moreover, in the subject group,symptoms such as dysphoria, pharyngalgia, acne, menstrual clots, xerostomia,feverish sensation in palms and soles, tinnitus and cardiopalmus werealleviated more remarkably than the control group, and the difference wasextremely significant (P<0.01). Regarding laboratory detection, in thesubject group, WBC, LYMN, HGB, PLT, ALB, C3, C4, CH50 rose notably after therapy,and the discrepancy was extremely significant (P<0.01). Furthermore, ERY,PRO, ANA, ESR and FIB fell remarkably, and negative conversion of anti-ds-DNAantibody was visible. The discrepancies were extremely significant (P<0.01)after treatment. The function of reducing ESR and converting anti-ds-DNAantibody to negative were better than the control group, and the differencewas significant (P<0.05). (3)Empirical study:①The effects of serum oftype-ⅣLN patients on GMC: Different concentration (5%,10%,15% and 20%)serumof type-ⅣLN patients could promote GMC proliferation more markedly comparedwith control serum, and the difference was extremely significant (P<0.01).The 15% serum of type-ⅣLN patients made the most significant impact.②Theeffects of traditional Chinese medicine of HCDP on GMC proliferation andapoptosis: Being intervened by the 15% serum of type-ⅣLN patients, differentconcentration (low, medium and high) serum containing traditional Chinesemedicine of HCDP could inhibit the proliferation of GMC and induce theapoptosis of GMC more notably than the control group, showing extremelysignificant difference (P<0.01).The function indicated an obviousdose-effect relationship. The effect of high concentration serum was the mostnotable compared with other groups, the discrepancies were extremelysignificant (P<0.01).③The synthesis of Bcl-2 protein and the expressionof Bcl-2mRNA in GMC: Being intervened by the 15% serum of type-ⅣLN patients,different concentration (low, medium and high) serum containing traditionalChinese medicine of HCDP could inhibit the synthesis of Bcl-2 protein and theexpression of Bcl-2mRNA, a gene inhibiting apoptosis, more remarkably thanthe control group. The function also showed an apparent dose-effectrelationship. The effect of high concentration serum was the most notable.Conclusions (1)The pathogenesis of LN is very complicated. Abnormalproliferation and apoptosis of GMC is an important mechanism. Western medical science-oriented therapy has disadvantages such as insensitivity of somepatients, many side effects, etc. Studying traditional Chinese medicine thatcan inhibit GMC proliferation and induce GMC apoptosis is tremendouslypromising in clinical application. (2)Clinical research shows: TraditionalChinese medicine of HCDP can relieve main symptoms of LN of toxic heat andblood stagnation, improve laboratory detection indices, and reduce SLEDAI andthe integral syndrome of TCM. The therapeutic effects are satisfactory andthe side effects are fairly few. (3)Empirical study discovers: The serum oftype-ⅣLN patients can remarkably promote the proliferation of GMC and the15% serum has the most notable impact. Being intervened by the 15% serum oftype-ⅣLN patients, the serum containing traditional Chinese medicine of HCDPcan inhibit the proliferation of GMC, induce the apoptosis of GMC, and inhibitthe synthesis of Bcl-2 protein and the expression of Bcl-2mRNA, a geneinhibiting apoptosis remarkably. This may be the molecular mechanism of TCMof HCDP curing LN and provides experimental and theoretical evidence forclinical application.

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