参莲消癥汤治疗中晚期肝癌临床与实验研究
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摘要
一.研究目的:
     观察中晚期原发性肝癌患者中医单证证型分布特点,初步探讨岭南肝癌的病因病机特点,为经验方“参莲消癥汤”(SLXZT)提供立方依据:观察“参莲消癥汤”对中晚期患者的临床疗效及对H_(22)肝癌荷瘤小鼠的基础药效作用以初步评价参莲消癥汤的临床有效性及可能的作用机理。
     二.研究方法:
     1.文献研究部分:综述了现代医学及古代和现代中医医家对原发性肝癌病因病机的认识及治疗的研究现状,在此基础上进行评述和展望。
     2.临床研究部分:将广州、江门两地纳入的76例中晚期原发性肝癌,观察单证证型分布特点;随机单盲分为参莲消癥汤(SLXZT)组42例和对照组34例,在同予西医对症支持治疗及斑蝥酸钠维生素B6治疗的基础上,中药组加服用SLXZT,用药1月后进行临床症状、病灶、生存质量、血分析、肝肾功能、凝血功能、甲胎蛋白(AFP)、T细胞亚群等变化的比较以评价疗效。
     3.实验研究部分:建立H_(22)肝癌荷瘤小鼠动物模型,雌雄各半,设立正常组、模型组、CTX阳性药物对照组及SLXZT低、中、高剂量组,每组10只,日常观察小鼠的一般生活状态,用药10日后观察药物对荷瘤小鼠抑瘤率、胸腺指数,流式细胞仪(FCM)检测脾脏T细胞亚群变化及肿瘤细胞凋亡率,放免法检测(RIA)血清细胞因子IL-2、THF-α水平及的影响。
     三.结果:
     1.文献研究部分:文献研究表明,目前原发性肝癌的病因及发病机制尚未完全清楚,治疗上多以综合性治疗为主。中医药参与的原发性肝癌多学科综合治疗模式成为临床疗效提高的关键及肝癌临床研究的热点之一。近年来应用中医中药和中西医结合治疗原发性肝癌,从药物抗癌机制的理论探索到临床疗效研究取得一定成果。
     2.临床研究部分:
     (1)广州、江门两地76例中晚期肝癌单证证型出现频率依次为:脾虚型(65.79%)、气滞型(51.32%)、血瘀型(48.68%)、湿证型(40.79%)、热毒型(34.21%)和阴虚型(18.42%),脾虚证为最常见的中医单证证型,其次为气质型和血瘀型,阴虚型较为少见;
     (2)对症状、生活质量改变的观察显示,在综合症状改善方面,治疗后SLXZT组和对照组在症状积分上均较治疗前显著下降(P<0.01),治疗后SLXZT组症状缓解率达54.76%,明显优于对照组29.41%(P<0.05)。在生存质量观察中,治疗后SLXZT组生存质量提高率为30.95%,优于对照组20.59%,但两组间在提高率、稳定率及降低率方面比较无统计学差异(P>0.05);
     (3)在稳定瘤体及对AFP的影响显示,SLXZT组治疗稳定率为69.05%,接近于对照组的64.71%,两组间无统计学差异(P>0.05)。治疗后两组AFP仍呈进行性的升高,但较治疗前无统计学差异(P>0.05),治疗前后两组间均无统计学差异(P>0.05);
     (4)对肝脏功能的影响显示,治疗后两组AST、ALT、TB较治疗前均有所下降,其中两组均以ALT下降明显,较治疗前具有统计学差异(P<0.01);而两组ALB、A/G较治疗前也均有下降,两组ALB与治疗前比较具有统计学差异(P<0.05);
     (5)对外周血T细胞亚群的影响结果显示,治疗后SLXZT组CD3、CD4、CD4/CD8均较治疗前显著性升高(P<0.01),CD8稍有下降但与治疗前无明显差异(P>0.05);对照组治疗后CD3、CD4、CD8、CD4/CD8与治疗前相当(P>0.05),CD3、CD4及CD4/CD8均低于SLXZT组,具有统计学差异(P<0.05);
     (6)两组在治疗前后血分析、肾功能及凝血功能等相关指标变化差异不大(两组间及两组治疗前后比较均无统计学差异,P>0.05)。
     3.实验研究部分:
     (1)祛邪作用①H_(22)肝癌荷瘤小鼠造模成功;②药物对抑瘤率的影响显示,SLXZT各组瘤重均低于模型组,CTX组抑瘤率为45.27%,SLXZT低、中、高剂量组均具有不同程度的抑瘤作用,其中以中、高剂量组作用显著,瘤重与模型组比较具有统计学差异(P<0.05)。中药三个剂量组的抑瘤率分别为25.75%、35.29%、41.58%,存在一定的量效关系(中药组间比较无统计学差异P>0.05);③对肿瘤细胞凋亡率的影响显示,模型组凋亡率为5.27%,CTX组为12.25%,SLXZT低、中、高组的凋亡率分别为7.50%、8.30%和8.58%,均显著高于模型组,具有显著性差异(P=0.000)。
     (2)扶正作用①对一般状况观察显示,中药各组小鼠在脱毛、活动力、毛发光泽等方面均优于模型组和CTX阳性对照组;②药物对胸腺指数的影响显示,CTX组胸腺指数最低,为27.29±6.03mg/g,与正常组比较具有显著性差异(P<0.05),SLXZT组的脾脏指数高于CTX组,但无统计学差异(P>0.05);③对荷瘤小鼠血清IL-2水平的影响显示,各组荷瘤小鼠血清IL-2水平较正常组均有不同程度的下降,以CTX组下降幅度最大,与正常组比较存在有显著性差异(P<0.05),中药各组小鼠血清IL-2水平有所回升,其中SLXZT高剂量组IL-2水平显著升高,接近正常组小鼠(P=1.000):④荷瘤小鼠TNF-α水平影响显示,SLXZT低、中、高剂量组荷瘤小鼠TNF-α水平高于正常组和CTX组(P<0.05),但低于模型组(组间比较P>0.05);⑤对荷瘤小鼠脾脏T细胞亚群的影响显示,与正常组比较,实验各组T细胞亚群呈紊乱状态,CD3、CD4低下,CD8升高,CD4/CD8比值倒置,其中以CTX组CD3、CD4下降最为显著(P<0.01,P<0.05)。应用SLXZT后,CD3、CD4、CD3/CD8升高,CD8有所降低。
     四.结论:
     本研究结果提示受岭南地域气候特点的影响,中晚期肝癌患者中医单证证型分布以脾气虚型最为常见,其次为:气滞型、血瘀型、湿证型,阴虚型相对少见。结合岭南肝癌以脾虚为本,气滞血瘀、湿毒蕴结为标的病因病机,针对性设立的以健脾理气、祛湿解毒、化瘀消癥法为代表的经验方“参莲消癥汤”可以改善患者综合症状并显示出一定的提高患者生存质量的优势,具有良好的细胞免疫调节作用。
     实验研究证实,参莲消癥汤具有直接抑制肿瘤、促进H_(22)荷瘤凋亡的祛邪作用及良好的对荷瘤小鼠免疫功能的调整作用。诱导细胞凋亡并通过调节和增强机体免疫功能可能是参莲消癥汤抗肿瘤作用机制。
1. Objective:Observed the medium or late stage primary liver cancer on the rules ofsingle syndrom, in order to discuss the etiology and pathogenesis characteristic of thesouthern China PLC, offered the foundations for Shenlian Xiaozheng Tang (SLXZT); Toobserve the therapeutic efficacy of SLXZT on medium or late stage primary liver cancerand the basic efficacy on H_(22) mice liver cancer, discuss the possible mechanism;
     2. Methods:
     2.1 Literature syudy: A literature review was carried out to summarize the westenmedcine and TCM etiology with pathogenesis of PLC,as well as its treatment. Meanwhile,based on the review results to discuss and expect.
     2.2 Clinical research: Collected 76 eligible patients from Gruangzhou and Jiangmen, toassess the single syndrome; 76 patients with stageⅡ-Ⅲliver cancer were dividedrandomly into two group, as SLXZT treated group A(42 patients) and control group B(34patients). The two groups accepted the same treatment with protecting liver, according tosyndroms and Sodium Cantharidinate Vitamin B6 Injection. Meanwhile, 42 patients inSLXZT treated group were treated with SLXZT more. The therapeutic efficancy of the twogroups were evaluated after curing one month.
     2.3 Experimental research:Copied the hepatocarcinoma 22(H_(22)) solid tumor-bearing micemodel, 30 male mice and 30 female mice were randomly divided into a normal group, amodel group, a CTX positive group, a low dosage of SLXZT, a moderate dosage of SLXZTand a high dosage of SLXZT, 10 mice in each group. The general condition, the inhibitoryrate of solid tumor and the thymus index were observed. T lymphocytes functions and thecell apoptosis was measured by Flow Cytometry(FCM). Radioimmunaoassay(RIA) examedthe content of IL-2 and TNF-αin serum.
     3.Results:
     3.1 Literature study: The results of literature review showed that the etiology withpathogenesis of PLC were not very clear. Generally comprehensive therapies are used totreat PLC. Multi-disciplinary comprehensive therapy combining with TCM has been the key to increase the curative effect and the hot topic of clinical study on cancer. In recentyears, some fruits on using integrated traditional and western medicine to treat PLC hasbeen achieved, which are not only about the mechanism of anti-cancer medicine but alsothe study on the curative effect.
     3.2 Clinical research:
     (1) The single syndrome distributing characteristics of the 76 patients were that spleendeficiency syndrome(65.79%), the most common syndrome in the patients.Othersyndromes in orders were Qi stagnation syndrome(51.32%), blood stasis syndrome(48.68%), excess of damp syndrome(40.79%), excess of heat syndrome(34.21%) andYin-deficiency syndrome(18.42%).
     (2) After curing one month, the two groups were better in symptoms thanpretherapy(P<0.01).The Group A (54.76%)had a better effect in the remission ofsymptoms than Group B(29.41%), P<0.05; The Group A (30.95%) had slightlyadvantage in improving the Kamofsky score than Group B (20.95%), but nosignificance in statistics(P>0.05).
     (3) About stabilizing the tumor, The Group A (69.05%)had a same effect with GroupB(64.71%), P>0.05; AFP still ascending after treatment, but there was no significantdifference among the two groups in the whole therapy course.
     (4) Comparing between the two groups, ALT, AST and TB descended than pretherapy, andthe descending of ALT has significant difference (P<0.01);However, ALB and A/G alsodescended, but have no significance in statistics than pretherapy.
     (5) The results of peripheral blood T lymphocytes functions showed that the number ofCD3, CD4 cells and CD4/CD8 were significant increased in Group A than beforemedcine (P<0.01) and have much better than Group B, CD8 had slightly decreased thanbefore medcine (P>0.05).
     (6) There were no significant difference in blood analyse, kidney functions and cruorfunctions between the two groups among the whole therapy.
     3.3 Experimental research:
     (1) The effect of eliminating pathogenic factors:①Mice models of the solid tumor livercancer were successfully established;②The tumor weights of each dosage of SLXZTgroup and CTX group were significant lower than model group, the inhibitory rate ontumor growth of CTX group was the lowest(45.27%). In the different dosage SLXZTgroups, the more dosage grew with the higher inhibition rate of minor.③The cellapoptotic rate of model group was 5.27%, each of the therapeutic groups wasstatistically higher than that of the model group (P<0.01), CTX group was 12.25%. In the different dosage SLXZT groups, the more the dosage grew, the higher the cellapoptotic rate was.
     (2) The effect of strengthening anti-pathogenic qi:①About general condition, the groupsdeal with SLXZT had less depilation phenomena and better activity than the CTX groupand model group;②The thymus index of CTX group was 27.29±6.03mg/g, whichstatistically lower than normal group (P<0.05), each of the herb groups was higher thanCTX group, but have no significance in statistics;③The results of serum IL-2showed that compared with the normal group, IL-2 reduced in every experimentgroup,CTX group has the lowest IL-2 level, each of the herb groups was higher thanCTX group, serum IL-2 level of the high dosage SLXZT group was close to normalgroup;④The content of serum TNF-αof every SLXZT group was higher than that ofthe normal group and CTX group, but lower than that of the model group;⑤Theresults of T lymphocytes functions showed that compared with the normal group,CD3,CD4,CD4/CD8 reduced in every experiment group, T lymphocytes functions wasfoul-up, SLXZT has the effect of increasing the content of CD3,CD4 and CD4/CD8.
     4. Conclusion:
     In the southern China, the special environmental and climate factors would influencethe etiology and pathogenesis of PLC. The single syndrome distributing characteristics ofthe 76 patients were that spleen deficiency syndrome(65.79%), the most common syndromein the patients.Other syndromes in orders were Qi stagnation syndrome(51.32%), bloodstasis syndrome (48.68%), excess of damp syndrome(40.79%), and the appearance oftheYin-deficiency syndrome(18.42%) was the least .According to the etiology andpathogenesis of PLC, which spleen deficiency is the principal aspect and qi stagnation andblood stasis, damp and toxic substances accumulation is the secondary aspect, ProfessorChen Ruishen established theapeutic methods was invigorate the spleen, replenishing qi,clear out damp, remove toxic substances, remove stasis and eliminate mass,named"Shenlian Xiaozheng Tang (SLXZT)". The results showed that SLXZT had a well effectin the remission of symptoms, in addition, it also had slightly advantage in improving theKamofsky score. SLXZT had a well effect on improving the cell-immunity.
     The results of experimental research showed that SLXZT had the effect of directkilling liver rumor in mice, inducing the apoptosis of the H22 liver cancer cells and wellimproving the immunity of mice,which would be the partial mechanism of SLXZT infighting the PLC.
引文
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