中文摘要

中文摘要

目的： 通过观察裴氏软肝消痞丸(PRGXP)对小鼠移植性肝癌H22瘤组织中P27、Bcl-2表达的影响，探讨裴氏软肝消痞丸对荷瘤小鼠免疫系统的调节、抑制肿瘤的作用，分析其治疗原发性肝癌的作用机制，为裴氏软肝消痞丸在临床上的应用提供理论依据。

方法： 通过小鼠右前腋部皮下接种法，建立荷H22（肝癌）实体瘤小鼠模型，随机平均分为空白组、模型对照组、裴氏软肝消痞丸小剂量组、裴氏软肝消痞丸中剂量组、裴氏软肝消痞丸大剂量组和复方斑蝥胶囊组。灌胃给药10d后，脊髓脱臼法处死，剥离肿瘤、胸腺、脾脏并称重，计算抑瘤率，测定胸腺指数(TI)及脾脏指数(SI)。采用免疫组织化学方法测定小鼠移植性肝癌H22瘤组织中P27及Bcl-2蛋白的含量。

结果：

1. 裴氏软肝消痞丸小、中、大剂量组的平均瘤重均低于模型对照组，分别为（0.935±0.227)g、(0.776±0.122)g、(0.926±0.237)g，与模型对照组比较均有统计学意义(p<0.05)。其抑瘤率分别为23.9%、36.8%和24.5%；
2. 裴氏软肝消痞丸小、中、大剂量组荷瘤小鼠的胸腺指数(TI)、脾脏指数(SI)均高于模型对照组，其中中剂量组TI为(53.2±15.6)mg/10g，较模型对照组增长31.4%，与模型对照组比较有统计学意义(p<0.05)；裴氏软肝消痞丸小、中、大剂量组SI分别为(58.9±14.5)mg/10g、(65.0±10.1)mg/10g、(55.2±15.9)mg/10g，与模型对照组比较有统计学意义(p<0.05)；
3. 裴氏软肝消痞丸小、中、大剂量组P27的阳性表达较模型对照组增强，与模型对照组比较均有统计学意义(p<0.05)；
4. 裴氏软肝消痞丸小、中、大剂量组Bcl-2的阳性表达均较模型对照组减少，与模型对照组比较均有统计学意义(p<0.05)。PRGXP中剂量组肿瘤组织中Bcl-2的阳性表达低于BM组，与BM组比较有统计学意义(p<0.05)。

结论： 裴氏软肝消痞丸具有抑制肿瘤生长的作用，可提高荷H22肝癌小鼠的胸腺指数、脾脏指数，通过提高P27蛋白的表达，减少Bcl-2蛋白的表达，起到抗肿瘤作用。

关键词： 裴氏软肝消痞丸；H22(肝癌)；P27；Bcl-2

裴正学系列方药的研究

ABSTRACT

Objective: Through observing the effect of Peishiruanganxiaopi pills (PRGXP) on the P27 and Bcl-2 in H22 tumor-bearing mice, we have discussed the regulating and antitumor effects of PRGXP on the immune system of tumor-bearing mice. To support for its clinical use, we aim to explore the basic mechanism of PRGXP's effect on primary carcinoma of the liver.

Methods: Firstly, we established the tumor-bearing mice model by hypodermic inoculation in the right front axilla of mice. All the mice were divided into blank group, model group, PRGXP low-dose group, PRGXP medium-dose group, PRGXP high-dose group, and Fufang Banmao capsule (BM) group. Ten days later, they were fed and then killed. Those tumors, thymuses and spleens were peeled off and weighted. The antitumor rate, the thymus index (TI) and the spleen index (SI) were calculated. By studying the expression of P27 and Bcl-2 protein in the tumor tissue by immunohistochemistry, we analyzed the statistics.

Results:

1. According to the dosage of the PRGXP, three groups can be divided: the low, the medium and the high. The weight of tumor in the three groups of PRGXP, separately is (0.935±0.227)g, (0.776±0.122)g and (0.926±0.237)g, which is lower than the model group (p<0.05) while the antitumor rate are 23.9%, 36.8% and 24.5%.
2. The TI and SI of the three groups are higher than the model group (p<0.05). The TI of the PRGXP medium-dose group is (53.2±15.6) mg/10g, and the percentage increase is 31.4% than the model group (p<0.05). The SI in low-dose group, medium-dose group and high-dose group of the PRGXP were (58.9±14.5) mg/10g, (65.0±10.1) mg/10g, (55.2±15.9) mg/10g, which indicated that the three groups were all statistical significances compared with model group (p<0.05).
3. The expression of P27 was increased in PRGXP groups and BM group, which were all statistical significances compared with model group (p<0.05).
4. The expression of Bcl-2 was decreased in PRGXP groups and BM group, which were all statistical significances compared with model group (p<0.05). The middle dose group of PRGXP has significant antitumor effect.

Conclusion: PRGXP could inhibit the growth of liver cancer, and could significantly increase the thymus index and the spleen index in H22 tumor-bearing. PRGXP was also shown significant antitumor effect through the increased expression of P27 and the expressions of Bcl-2 reduced.

Keywords: Peishiruanganxiaopi pill; H22 (transplanted liver cancer); P27; Bcl-2

前言

原发性肝癌（简称肝癌）是发生于胆管细胞和肝细胞的临床上常见的恶性肿瘤。《WHO 2008》

裴正学系列方药的研究

《世界癌症报告》将原发性肝癌列为十大恶性肿瘤之一，其居世界第5位，死亡率居第3位。肝癌的发病率在全球范围内逐年增长，全世界新发病例约62万人/年，其中55%在我国，并占我国肿瘤相关死亡第2位[1][2]。肝癌起病隐匿，初期症状不明显，病情发展快，恶性程度高且容易复发，5年生存率低，严重威胁人类健康。

肝癌有手术、化疗、放射、微创、生物、原位肝移植等多种治疗方式。手术是早期肝癌首选的治疗方法，但由于大部分患者就诊时，已经失去了手术机会，且由于患者对放疗的耐受性低、化疗后不良反应严重、肝癌对化疗药物敏感程度较差等原因，治疗效果不甚理想。我国的特色中医药，在治疗肝癌方面取得了较好的成绩。在肝癌早期，采用中医药治疗，可提高患者对手术的耐受性；手术后可以增强患者体质，清除残余病灶；放化疗的患者，可以减轻其毒副作用，提高疗效；长期服用中药，可以有效地防治肝癌复发和转移，改善患者生活质量，延长生存期。因此，以中医药为主对肝癌的综合防治，日渐被人们所重视。

根据肝癌患者肝脏肿大，肝区疼痛，黄疸，肝硬化征象等临床表现，肝癌的中医病名归属于"积聚"、"肝积"、"黄疸"、"鼓胀"等范畴。《诸病源候论》曰："痞气，肝病传脾，脾当传肾，肾冬适王，旺者不受邪，脾欲复还于肝，肝不肯受，故留结为积"；《济生方·瘤瘤积聚门》曰："肥气之状，在左胁下，大如覆杯，肥大而似有头足，是为肝积"。肝癌的中医病因病机主要为正气不足，邪气盘踞，因此肝癌属于正虚邪实之症。《医宗必读》曰："积之成也，正气不足，而后邪气踞也"。肝癌中医治法治则主要有健脾益气、清热解毒、补益肝肾、活血化瘀、化痰利湿等。

裴氏软肝消痞丸是我国著名中西医结合专家裴正学教授，在四十余年临床实践中总结的治疗肝癌的有效方药，全方主要由丹参、黄芪、柴胡、白芍、莪术、海藻、皂角刺、白花蛇舌草、半枝莲等药物组成，具有疏肝理气、软坚散结、破血祛瘀、清热解毒之功效，扶正祛邪，标本兼治。裴氏软肝消痞丸在对原发性肝癌、胃癌的治疗上，在减轻放、化疗引起的不良反应、提高患者生活质量等方面取得了满意的疗效。

本实验运用动物实验，通过免疫组化、图像分析等方法，对使用裴氏软肝消痞丸后的H22荷瘤小鼠抑瘤率、胸腺、脾脏指数进行计算与分析，从细胞增殖与细胞凋亡两方面检测P27及Bcl-2表达情况，研究裴氏软肝消痞丸抑瘤的作用机制，为裴氏软肝消痞丸在临床上的广泛应用提供理论依据。

立题依据