Toll-like Receptor Pathways To Explore The Mechanism Flavored The Campanulaceae Soup Treatment Of Endotoxin Induced Acute Lung Injury

Acute lung injury(abbreviated as ALI) refers to acute progressive respiratory failure with stubborn hypoxemia induced by different predisposing factors except cardiogenic cause. The pathogenesis of ALI is anfractuosity and is not completely elucidated by now, so there is no satisfactory therapeutic measure, which case fatality is 50%. According to its clinical manifestations, ALI belongs to the Pyretic pulmonary syndrome, the Syndrome of Dyspnea, or Thoracic Retention of Traditional Chinese Medicine(abbreviated as TCM). In the past few years, according to the new research of ALI about its pathogenesis, TCM researchers combinated methods of differential diagnosis of diseases with differentiation of symptoms and signs to treat ALI basing on the theory of Pyretic toxicity, but not lowered the fatality of ALI. So it was necessary to lucubrated the the TCM nosogenesis of ALI, and use the mechanism to direct our to find out the effective prescription and drugs.Now, datas indicate that 70% ~80% pathogenic bacterium causing ALI is gram-negative bacterium, which major antigen and pathogenicity ingredient is lipopolysaccharide (abbreviated as LPS) and the Toll-like receptor 4(abbreviated as TLR4) is the transmembrane signal transduction receptor of LPS. So the signal transduction pathway of LPS mediated by TLR4 should play an important role in the development of ALI. The theory also made us change the therapeutic strategy from diseases to sick person and think highly of the relation between the physiological functions of TCM lung with ALI. So we proposed that impairment of dispersing and descending function of the lung, vicious toxicant stagnated in the lung and the confused activities of Qi was the TCM nosogenesis of ALI. And then, the therapeutic principle named as Xuanfei Jiedu(abbreviated as XFJD) was in conformance with the physiological functions of the lung and in line with the pathogenesis of ALI. Therefore, the Decoction of Radix Platycodi with addition (or Jiawei Jiegeng Decoction, abbreviated as JWJGD) guided by the XFJD principle maybe have a satisfaction therapeutic effect on the ALI induced by LPS.Objectives1. To explore the role of LPS signal transduction pathway mediated by TLR4 in the nosogenesis of ALI2.To explain the relations between physiology and pathology of Lung with ALI and investigate the modern biology foundation about the pathomechanism of ALI and Visceral figure theory of Lung.3.To discuss the feasibility of the principle named as Open the inhibited lung-energy to detoxify nonage (or XuanFei Jiedu, abbreviated as XFJD) to treat ALI.4.To study the pharmacological mechanism of JWJGD from the holism, cell and molecular level to offer experimental groundwork to scientifically prevent and cure ALI.Methods1 Literatures StudyBasing on the relation between the Visceral figure theory of Lung with ALI and the outline of Symptom complex and treatment about the method named as opening the inhibited lung-energy of Zhang Zhongjing to explore the feasibility of XFJD, and discuss the theory foundation of JWJGD to treat ALI.2 Empirical Study2.1 Pharmacodynamics Study of JWJGD90 male Sprague-Dawley rats were randomly divided into 4 groups, 9 rats in normal group, 27 rats in model control group, 27 rats in Qingre Jiedu group, 27 rats in XFJD groups. Rats in the Qingre Jiedu group and XFJD group were intragastric administration with Huangqin Qingfei Decoction(composed with Scutellaria, Gardenia florida, et al.) and JWJGD(composed by Huangqin Qingfei Decoction added Jiegeng Decoction) respectively, the other two groups were intragastric administration with equal volume isotonic Na chloride for 6 days. On the seventh day, rats in the model group, Qingre Jiedu group and XFJD group were injected with a single dose of LPS(4.9mg/kg) by vena femorali, and the normal groups were injected with equal volume N. after that, observing and recording the symptoms of all the rats. Rats of the other three groups except the normal group were killed randomly on the 2h, 4h and 6h after injected with LPS. Three rats of each group were irrigated by left ventricle with 4% paraform to fix, and then making them into light microscope sample, the other 6 rats were killed after anesthetized and open the thorax to obtain the lung, throwing the lung into the liquid nitrogen after weighed. We evaluated the therapeutic effect of Huangqin Qingfei Decoction and JWJGD about ALI by measuring the lung coefficient, lung water content, MPO activity of lung tissue homogenate.2.2 Mechanism Study of JWJGDThe grouping and model-made methods were same with the pharmacodynamics study. Using the biochemistry method to measure the content of SOD and MDA of lung tissue homogenate and the content of NO, NOS and iNOS of serum; Using the ABC-ELISA method to measure the content of TNF- , IL-1β, IL-6, and IL-10 of serum; NF-κB p65 of lung was detected with Western-blot and immunohistochemical method; Using RT-PCR method, the TLR4 mRNA of lung was detected.Results1 Theoretic StudyThere is a close relation between the physiological functions of TCM lung with the development of ALI. Impairment of dispersing and descending function of the lung caused by Vicious and poison invasion is the pathology foundation about ALI; Struggling between the healthy Qi and pathogen, continue accumulation of patho-products always causes the vicious poison to obstruct the lung collaterals and disorder the activities of Qi, which all can aggravate the ALI; The lung cannot dominate Qi that is the key point of the turnover about ALI. So the XFJD method is an important way to treat ALI, and the JWJGD will have a satisfactory therapeutic effect on ALI.2 Experimental Study1.PMNs'assembling and adherency in lung is the premature event during process of ALI induced by LPS, PMNs can result in widely damages of lung vascular endothelial cells and alveolar epithelial cells, and the later can lead pneumonedema and micro thrombus;2.Imbalance between Oxidation and Antioxygen always occurs in ALI induced by LPS;3.There is imbalance between aggressive inflammatory response with anti-inflammatory response in the process of ALI induced by LPS, quickly and generous expression of TNF-,IL-1βis molecule mark of mononuclear macrophage activator, and the generous expression of IL-10 during the nonage of ALI always prognosticates immune function disorder;4.In ALI induced by LPS, NF-κB p65 always transfers from cytolymph to nuclear, the activation of NF-κB p65 has a kernel effect on the development of ALI;5.TLR is not only the first line to defense the exogenous evil invasion, but also the bridge between hereditary immunity with acquired immunity. During the process of ALI induced by LPS, generous mRNA gene expression of TLR4 is an important trigger point to aggravate the acute inflammatory reaction network;6. The followings are the mechanism of action of JWJGD to treat ALI induced by LPS:(1)Decreasing the aggregation and adherency of PMNs in the lung to attenuate the soakage in lung tissue;(2)Ant oxidation, cleaning oxygen free radical, and regulating the imbalance between Oxidation and Antioxygen;(3)Diminishing the content of TNF-,IL-1βand IL-10 in serum of ALI rats;(4)Inhibiting the transfer of NF-κB p65 from endochylema to nuclear to attenuate the expression of inflammatory factor;(5)Inhibiting TLR4 mRNA gene expression and the signal transduction pathway to regulate the activation of NF-κB.Conclusions1 Scientific basis of using XFJD principle to treat ALI. Basing on the relation between the Visceral figure theory of TCM Lung with ALI, we considered that the pathology location is lung, vicious and poison invading the lung resulted to impairment of dispersing and descending function of the lung was the pathogenesis juncture, which can lead to transit disorder about QI, blood and body fluid, so the fever vicious, water, phlegm and haemostasis, et al.suddenly accumulated, obstructing the lung collaterals to make activities of Qi confused, these were the core bit to aggravate ALI. Asthenia of healthy energy and magnificent of vicious leaded to the lung not dominating Qi and regulatory function disorder was the key point of the turnover about ALI. So the TCM nosogenesis of ALI indicated that physiologic function of TCM lung had an important role in the process of ALI. It also evocated the early treatment principle named as XFJD method. On the whole, the XFJD principle not only breaked through the traditional treatmental theory which was heat-clearing and detoxicating method, but also was the inheritance and development about Open the inhibited lung-energy to eliminate pathogen of Zhang Zhongjing.2 Modern biological foundation between some pathomechansism of ALI with the Visceral figure theory of TCM LungThere were many kinds of pathology action such as protein and lipoid dysbolismus, over expression of oxygen free radical and inflammatory factor and apoptosis, et al. We should apply combined and multitarget therapy to treat ALI induced by LPS because of its complicated pathology component. The research about LPS signal transduction pathway mediated by TLR4 can not only provide cut-in point for clinical control the cascades inflammatory reaction, but also partly reveal the biological foundation between macro theory such as Study on the Correlation Between Sluggishness of Lung-Wei, Diseases Caused by Vicious Factors and Lung regulating visceral activities with Toll-like receptors passageway and construct the bridge communicating the macro pathogenesis and micro pathogenesis about ALI.3 Pharmacological mechanism of action about JWJGD on ALIImpairment of dispersing and descending function of the lung, vicious factors obstructing the lung collaterals and activities of Qi disorder were the core pathology link of ALI, and the vicious factors was the important factors caused ALI, So we should open the inhibited lung-energy to detoxicate on the early process of ALI to interrupt the development of ALI. Using the method as opening the inhibited lung-energy to detoxicate and detoxicating to open the inhibited lung-energy to recover the physiologic function of lung. On the other hand, the synergy of complex prescription compatibility can be reflected in different patho-link and different target biological effect. JWJGD can significant ameliorate pneumonedema of ALI, regulate the imbalance between aggressive inflammatory responses with anti-inflammatory response, et al. all these may be the pharmacological mechanism of JWJGD to treat ALI induced by LPS.