【作者】 王三虎；

【导师】 王宗仁；

【作者基本信息】 第四军医大学 ， 中西医结合临床， 2007， 博士

【摘要】 肺癌是临床最常见的恶性肿瘤之一。但目前西医和中医的疗效都不尽如人意。其原因首先是肺癌究竟属于中医什么疾病概念不清,以往只是简单罗列,认为属肺积、痞癖、咳嗽、咯血、胸痛等范畴。本文提出了“肺痿的内容基本符合现代医学有关肺癌的论述”这一观点。论证了肺痿的咳嗽、咽喉不利、浊唾涎沫、气急、喉中水鸡声、咯血、血痰、胸痛、发热等症状都与肺癌的临床表现一致。将肺癌分为气阴两虚,痰浊泛肺;肺中虚寒,痰蒙清窍;痰粘胸咽,气机不利;阳虚痰壅,肺失宣降;表寒内热,肺气上逆;水积肺痿,正虚邪实;肺肾两虚,摄纳无权;气血双亏,阴阳俱损;其他型等9个证型。并为每个证型制定了治法和代表方剂。其中气阴两虚,痰浊泛肺和水积肺痿,正虚邪实2个证型的代表方剂润肺散结汤、葶苈泻水汤是笔者在经方的基础上结合自身临床经验新拟的。通过235例原发性支气管肺癌的临床观察,明确了原发性支气管肺癌中,气阴两虚,痰浊泛肺型占所有病例的30.64%,和另外8个证型均有显著差异(P<0.01)。说明气阴两虚,痰浊泛肺型是原发性支气管肺癌的最常见证型。就临床出现率多少排序,依次还有痰热粘滞,气机不利、顽痰壅滞,肺失宣降、气血双亏,阴阳俱损、水积肺痿,正虚邪实、肺肾两虚,摄纳无权、肺中虚寒,痰蒙清窍和表寒内热,肺气上逆。从病程来看,气阴两虚,痰浊泛肺这一证型贯穿始终,而气血双亏,阴阳俱损、肺肾两虚,摄纳无权、肺中虚寒,痰蒙清窍则明显集中在3期、4期,提示病久正气耗伤,阴损及阳的经典论述是有临床指导意义的。在22例水积肺痿,正虚邪实型肺癌中,腺癌有19例,占86.36%,远高于鳞癌的13.64%。提示临床对于病理报告为腺癌的肺癌患者要重视健脾宣肺利水化湿等治法。通过对72例气阴两虚痰浊泛肺型非小细胞肺癌进行的系统观察,证明润肺散结汤合并化疗,临床疗效可靠,主症好转率治疗组明显高于对照组,均显示P <0.01;治疗后,病灶均有缩小,缓解率(CR+PR)治疗组(55.55%)与对照组(50.00%)无显著性差异(P>0.05),而(CR+PR+NC)治疗组(91.66%)明显优于对照组(72.22%)(P<0.05):生活质量的改善和稳定方面,对照组也明显优于对照组,(P<0.05);毒副反应发生率,治疗组均低于对照组,差异有显著意义,P均<0.01;在完成化疗方面,治疗组也明显优于对照组,P<0.05);治疗组1年、2年、2年6个月的生存率均高于对照组,(P<0.05)。说明润肺散结汤合并化疗是治疗气阴两虚痰浊泛肺型非小细胞肺癌的有效方法。为观察润肺散结胶囊(RFSJC)对荷瘤小鼠(Lewis肺癌)的抑制作用。采用动物移植性肿瘤实验法,腋下接种Lewis肺癌,随机分组,口服RFSJC后观察瘤重变化和抑瘤率,并在光镜和电镜下观察瘤组织病理变化。以免疫组化检测肿瘤组织中Bcl-2和Bax蛋白的表达,并以流式细胞仪分析脾细胞内T淋巴细胞的比例的方法。结果:RFSJC用药组的抑瘤率为分别为36.5%、35.0%和30.1%;实验组与对照组相比,瘤质量和瘤体积也有显著差异(P<0.01)。瘤组织HE切片见明显的炎细胞浸润和明显的出血带,并有大片坏死区域,间质血管较少,电镜示凋亡小体出现。用药组Bcl-2蛋白的表达明显减少、染色变弱,IODT值显著减小(P<0.01),用药后Bax蛋白表达强度与对照组相比无明显差异。RFSJC各剂量组CD3+、CD4+和CD8+ T细胞比例均明显增高(P<0.01)。说明润肺散结胶囊对Lewis肺癌小鼠瘤体有明显的抑制作用,其机制可能与诱导肿瘤细胞凋亡和提高机体免疫力有关。为观察润肺散结胶囊对体外培养的鼠Lewis肺癌细胞的抑制作用。我们将不同浓度含润肺散结胶囊溶液的血清加入体外培养的Lewis肺癌细胞,用MTT法测定体外抗瘤活性,用流式细胞仪、免疫组化等观察其对肿瘤细胞周期及细胞凋亡的诱导作用。结果:润肺散结胶囊含药血清处理Lewis肺癌细胞24 h后,细胞形态出现典型凋亡特征,当浓度高于16μg/mL、作用72 h时,Lewis肺癌细胞的存活率低于5%,其IC50约为0.5μg/mL;经RFSJC处理后,癌细胞增殖能力明显下降;经RFSJC作用36 h后,S期Lewis肺癌细胞比例在整个细胞周期所占百分比较对照组明显增高(P<0.05),尤其是4.5μg/mL的Lewis肺癌细胞处理后S期比例升高至40.1%,G0/G1期细胞有所减少,M/G2期细胞较未经Lewis肺癌细胞处理的对照组也有减少,尤其是中剂量组,M /G2细胞所占比例极低;经RFSJC处理后,凋亡细胞比例显著增加;用药后也出现p53、Bcl-2阳性表达细胞比例显著下降(P<0.01),c-myc阳性着染细胞比例明显升高,着色变深。结论:润肺散结胶囊对体外培养的Lewis肺癌细胞有明显的抑制作用,肿瘤细胞S期比例的升高提示其作用的环节可能为DNA合成受阻,其机制可能为诱导肿瘤细胞凋亡。更多还原

【Abstract】 Primary pulmonary cancer which can simply called lung, is the most common malignant tumor. However there is no good effective treatment whether in Chinese medicine or western medicine. The main reason in Traditional Chinese Medicine theory, the concept of pulmonary cancer is usually confused, it is always just simply listed in numerous names in the past, such as Pulmonary mass, Hypochondriac lump, Cough, Hemoptysis, Blood spuyum, Chest pain, Fever. The author comes up with a new viewpoint that the symptom of Consumptive is basically accord with lung cancer. In this research explore and prove that cough, laryngopharyngeal paresthesia, ptyalism, short breath, wheezing sound, hemoptysis, blood sputum, chest pain, fever and other clinical manifestations in consumptive lung disease are similar to lung cancer. Lung cancer can be divided into the following 9 types: deficiency of both qi and Yin, stagnation of phlegm in the lungs, cold deficiency in the lungs, stagnation of phlegm in the interior, accumulation of phlegm-heat, disorder of qi, Yang deficiency due to phlegm stagnation, impairment of the descending and purifying effects of the lung, exterior heat and interior cold, upward adverse flow of the qi, phlegm retention due to consumptive lung disease, weak body resistance resulting in existence of pathogenic factors, deficiency of both the lung and the kidney, regulation disorder, deficiency of both qi and blood, deficiency of both Yin and Yang patterns and so other forms. And formulate treating methods and kepresentative prescription for every syndrome, especially deficiency of both qi and Yin, stagnation of phlegm in the lungs, and phlegm retention due to consumptive lung disease, weak body resistance resulting in existence of pathogenic factors. The two types of phlegm retention due to consumptive lung disease, weak body resistance resulting in existence of pathogenic and deficiency of both qi and yin, stagnation of phlegm in the lungs, whose kepresentative prescription were Runfeisanjie Decoction and Tinglixieshui Decoction which were made from classical medical books combined with clinical experience by writer.Through the clinical observation on 235 cases primary brouchogenic caranoma of lung, we can definitely describe the lung cancer. The type of deficiency of both qi and Yin, stagnation of phlegm in the lungs constitute 30.64% in the whole that have marked differences with other 8 types. P<0.01, which prove that primary brouchogenic caranoma of lung is the most common type. According to the appearance rate, also have the type of accumulation of phlegm-heat, disorder of qi, pertinatious phlegm stagnation, impairment of the descending and purifying effects of the lung, deficiency of both qi and blood, deficiency of both Yin and Yang, phlegm retention due to consumptive lung disease, weak body resistance resulting in existence of pathogenic factors, deficiency of both the lung and the kidney, regulation disorder, cold deficiency in the lungs, stagnation of phlegm in the interior, exterior heat and interior cold, upward adverse flow of the qi, phlegm retention due to consumptive lung disease. As for course of disease, the type of deficiency of both qi and Yin, stagnation of phlegm in the lungs from the beginning to the end, but deficiency of both qi and blood, deficiency of both Yin and Yang, deficiency of both the lung and the kidney, regulation disorder, cold deficiency in the lungs, stagnation of phlegm in the interior were centralized in 3 and 4 period, pointed out healthy qi consumed because of long disease, deficiency of Yin affecting Yang also have clinical leading meaning. There are 22 cases of phlegm retention due to consumptive lung disease, weak body resistance resulting in existence of pathogenic factors in the lung cancer. 19 cases adenocarcinoma constitute 86.36%, higher than squamous carcinoma 13.64%. We should focus on strengthening the spleen and promoting the dispersing functions of the lung, inducing diuresis and resolving dampness if pa thological reported the lung cancer patient caused by adenocarcinoma.But the systematic observation on 72 cases with the type of deficiency of both qi and Yin, stagnation of phlegm in the small cell lung cancer (SCCL), which proved that Runfeisanjie Decoction merged chemotherapy have reliable clinical curative effect, principle syndrome improvement rate high than control group P<0.01.Focus also shrinked, there is no greatly differences with remission rate (CR+PR) between treating group(55.55%) and control group (50.00%) after treatment (P>0.05),but (CR+PR+NC) treating group (91.66%) higher than control group (72.22%) (P<0.05), even from the quality of life improvement and stabilization aspects (P<0.05); as for toxic and side reaction rate, treating group lower than control group that have greatly differences<0.01. In the period of finishing chemotherapy, treating group higher than control group with survive rate in 1 year,2year,6 months. So, Runfeisanjie Decoction plus chemotherapy is an effective method for treating the type of deficiency of both qi and Yin, stagnation of phlegm in the lungs non-small cellular.To observe the effect of Runfeisanjie Capsule on inhibiting mouse Lewis Lung Carcinoma tumor growth. A piece of Lewis Lung Carcinoma was transplanted into the subcutaneous space on the armpit of each C57BL mouse. Mice were random separated into several groups. After orally administered, the mean weight of the subcutaneous tumors was examined, then the average tumor control rate was analysed. And observed the change of organization of tumor by light microscope and electron microscope, Bcl-2 and Bax were detected by the method of immunity histochemistry and the proportion of T lymphocyte were calculated by flow cytometer FCM. In the group treated by RFSJC, the tumor control rate was 36.5%, 35.0% and 30.1% respectively. Compared to the tumor model group, the weight and the volume of tumor were decreased significantly (P<0.01). Cells of inflamed infiltration in HE dyeing and hemorrhage band, a piece of necrosis zone, blood vessel less, apoptotic body appeared in electron microscope. The express of Bcl-2 and the value of IODT decreased greatly (P<0.01). The express of Bax changed little. The proportion of CD3+, CD4+ and CD8+ increased greatly (P<0.01). Being proved these results suggest that RFSJC could inhibit the growth of mouse Lewis Lung Carcinoma, and its mechanism may be related with its action of inducing the apoptosis of tumor cells .To investigate the effect and the mechanism of Runfeisanjie Capsule (RFSJC) on Lewis pulmonary tumor in vitro, we treated the Lewis cells with RFSJC plasma, the activation of anticancer were examined by MTT assay. The rate of inhibition and life prolongation were calculated and pathological sections were observed. Apoptosis of tumor cells was detected by flow cytometry and immune histochemistry after being treated with RFSJC. RFSJC inhibited Lewis cells proliferation significantly at the concentration above 16μg/mL for 72 h and its IC50 was 0.5μg/mL. The shape of cells appeared typical characteristic of apoptosis. The percentage of apoptosis increased significantly treated by RFSJC. After intervene by RFSJC 36hour later, the apoptotic percentage of Lewis lung cancer higher than control group in S period, (P<0.05), especially the group of 4.5ug/mL, S period of Lewis lung cancer amount to 40.1%, while cells number decreased in G0/G1 period as well as M/G2 period which compared to processing Lewis lung cancer, especially middle dosage group, proportion in M/G2 period was very low. It was thus evident that proportion of positive expression for p53 and Bcl-2 decreased obviously after medication, and the proportion of c-myc increased obviously (P<0.01) and color darkened. Result: RFSJC could inhibit the growth of pulmonary tumor cultured in vitro and it’s antitumor mechanism may be explained that the cancer cells in S period of is accumulate show that the key link may be the blocking of the DNA synthesis.更多还原