培土生金法通过TLR4信号通路干预LPS诱导的肺癌增殖研究

The proliferation of tumor cells is the result of its interaction with the microenvironment.chronic infection combined with inflammation promote tumor development and growth of metastases by up-regulating angiogenesis and increasing the secretion of carcinogenic cytokines .LPS via TLR4 leads to activation of many transcription factors,including NF-κB and AP1，as well as regulation of expression of inflammatory factors , which may initiate and promote tumorigenesis .Clinical studies have shown that there was a spectacular therapeutic index on the treatment of lung cancer with the method of "Reinforcing earth to strengthen metal".Our recent findings exhibit that the LPS encourages SPCA1 cells progression, and the medicated serum of Reinforcing earth to strengthen metal can inhibit LPS-induced proliferation of SPCA1 cells; Previous experimental and clinical evidence of our group demonstrated that inflammatory activity in lung tissue of asthmatic rat models could be reduced by regulating TLR4/NF-κB pathway with the " Reinforcing earth to strengthen metal "strategy.A hypothesis was,thus, presented by our research group that the therapy of lung cancer by the use of "Reinforcing earth to strengthen metal " leads to a reduction of metabolism and proliferation rate of the cells based on the TLR4 signaling pathway associated to the inhibition of LPS-induced inflammatory response and modulating process of angiogenesis,suppressing the secretion of carcinogenic cytokines and induction of tumor cell apoptosis.Aim to Test this theory,we observed the effect of LPS on lung cancer, TLR4 signaling pathway and the Regulatory role of method of " Reinforcing earth to strengthen metal", which reveals the impact of the inflammation in the carcinogenic process of lung cancer.This article is open up new avenues and strategies for the Prophylaxis and treatment of lung cancer by compound herbal medicinal Prescription which was composed by the rule"Reinforcing earth to strengthen metal ".

肿瘤细胞的增殖是其与微环境相互作用的结果。慢性感染和炎症通过促进血管生成和促肿瘤因子分泌等促进了肿瘤的发展。LPS能够通过TLR4激活NF-κB和AP1等转录因子，从而促进癌症发生、发展。临床研究表明培土生金法在肺癌的治疗中具有显著疗效。本课题组研究表明LPS能够促进肺癌SPCA1细胞的增殖，而培土生金法含药血清能够抑制LPS诱导的SPCA1细胞增殖；培土生金法能够通过调控TLR4/NF-κB通路抑制哮喘大鼠肺组织的炎症反应；但是其是否能通过TLR4信号通路抑制肺癌细胞的炎症反应还有待与进一步研究。因此本课题组提出培土生金法能够基于TLR4信号通路，抑制LPS诱导的炎症反应，调控炎性血管的生成、促肿瘤因子的分泌及肿瘤细胞凋亡等，从而抑制肺癌发展的假说。为证实该假说本文通过观察培土生金法对LPS诱导的肺癌增殖及TLR4信号通路变化的影响，从而揭示培土生金法抗炎症反应抑制肺癌发生、发展的机制。

中医学认为中焦脾胃虚弱，正气虚损是肺癌发生的根本原因，培土生金法用补脾益气的方药补益肺气的方法防治肺癌，充分体现了中医的整体观。肿瘤细胞的增殖是其与微环境相互作用的结果。慢性感染和炎症通过促进血管生成和促肿瘤因子分泌等促进了肿瘤的发展。LPS能够通过激活TLR4信号通路，从促进癌症发生、发展。本课题组采用采用体内与体外实验相结合的方法，运用分子生物学、血清药理学等技术，以肺癌小鼠模型和体外肺癌细胞株为研究对象，系统研究了培土生金法对肺癌增殖过程中TLR4信号转导通路的调控机制。体内研究表明培土生金法复方能够降低肺癌小鼠模型TLR4、VEGF、抑制凋亡基因bcl2的表达，增加促凋亡相关基因表达bax、caspase3表达，从而促进肿瘤细胞凋亡和血管生成；体外实验研究表明LPS能够促进肺癌SPCA1细胞的增殖，而培土生金法复方中药能够通过TLR4信号通路抑制LPS诱导的肺癌细胞增殖，抑制肺癌细胞的炎症反应，调控炎性血管的生成、及肿瘤细胞凋亡等。TLR4-siRNA能有效沉默肺癌细胞中TLR4的表达，能够阻滞细胞的生长，抑制细胞的增殖，在促进细胞凋亡的同时能够影响细胞周期分布从而抑制肿瘤细胞的生长，特异性干预TLR4基因的表达能够有效阻断LPS引起的炎症反应，从而影响肿瘤的发生与进展，有可能成为治疗肺癌的一种新手段。本课题为揭示培土生金法抗炎症反应抑制肺癌发生、发展的机制提供了新的思路与方法。为基于中医药理论发掘传统的中药复方治疗肺癌，并探索其作用的机制，具有重要的实际应用意义。