| Pulmonary interstitial fibrosis (PF) is a kind of disease characterized by diffusion, infiltration and fibrosis as a common outcome of many interstitial lung diseases. Constantly, the course of PF progressively develops. Its pathological features are the chronic inflammation leading to persistent injury of alveolar cell, over reproduction of lung fibroblast (FB) and excess deposition of extracellular matrix (ECM). It′s well known, the injury of tissues is usually accompanied by inflammation and repairing course. If the scope of injury is small, tissues can recover to normal structure and function, such as the infection reducing by most of bacteria and virus. Otherwise when the range is relatively big or recurrence, the frequent repairing will lead to fibrosis or scar formation, which will influence flexibility and respiratory function and induce the patient died of heart and lung failure as well as hypoxemia. As result of its pathogenesis is not thoroughly clear so far, there is no specific medicine in clinic. The traditional medicine is mainly glucocorticoid and immunosuppressant, delaying the course of PF through relieving inflammation. But to the patient diagnosed as PF in clinic, not only fibrosis has been developing and the disease condition had been developing continuously, but also the curative effect is still dissatisfactory and obvious adverse reactions were induced after long application. Therefore at present, the urgent question is to develop the better effect and high security medicine for PF. PF is mostly divided into the category of"Consumptive Lung Disease""Bi syndrome of the lung"in TCM, believed that the location of PF is in lung, also relating to multiple viscera, and its pathogenesis due mostly to deficient vitality and excessive evil. Through combining with the study on the ancient and modern literature and the long clinical observation, we considered that this disease is belonged to the category of"Consumptive Lung Disease", and the ill location lies in lung, but closely connective with spleen, and the key of pathogenesis is deficiency and stasis, so we put forward the hypothesis of deficiency of spleen and blood stasis. Furthermore combining with the theory of TCM, we also believes that invigorating spleen and lung, promoting blood and stasis-resolving is the basically big method, and applies the modification of the herbal compound of strengthening earth to support metal and stasis-resolving in treating PF, and has achieved the satisfactory effects in clinic. In order to further explore the effect of the herbal compound on PF, we apply the compound to treat PF rats, choose relative molecular biological markers, and use advanced experimental technique, observing the effect of the herbal compound on PF rats from different levels and angels, which provide theory basis for treating PF in clinic.In this experiment, we apply BLM to induce the PF model on rats, studying the effect of"strengthening earth to support metal and stasis-resolving compound"on PF rats, at the time of 7d,14d and 28d, exploring the mechanism of the herb from the different levels and angels such as molecular biology, immunity and pathology etc,so as to find the more effect medicines for the clinic. The experimental research consists of four parts as following:1 The effects of"strengthening earth to support metal and stasis-resolving compound"on pathomorphology and lung index in PF rats.Through observing general status of PF rats (such as weight, movement and respiration etc), Lung index and the changes in pathomorphology (HE and MASSON stain), studying the effects of"strengthening earth to support metal and stasis-resolving compound"on PF rats, and then comparing the effects of the herbal compound according to statistic data.The results showed as following: Exclude the normal group, other groups in PF rats had the phenomena of weight reducing, movement decreasing, less eating and lung index increasing. Under the microscope, alveolitis and collagen deposition were observed in different degrees among lung interstitial by HE and MASSON stain. After the herbal compound treatment, the general status of PF rats were improved, lung index reduced, and alveolitis and fibrosis relieved on pathomorphology.The results:⑴The PF rat model induced by BLM was successfully duplicated.⑵The herb compound of"strengthening earth to support metal and stasis-removing"can improve the degree of the alveolitis and fibrosis in PF rats.2 The study on the effects of"strengthening earth to support metal and stasis-resolving compound"on contents of HYP in PF rats.Though the research on the effects of"strengthening earth to support metal and stasis-resolving compound"on the contents of HYP in PF rats, discussing the change about contents of HYP in the process of PF. HYP is one of the main components in the collagen of body, and very few in the elastin, whereas no in the other protein. Therefore, HYP is the endemic components of collagen fiber. So that it is the diagnostic marker of PF and can judge the degree of PF.The results showed as following: Besides the normal group, the deposit of collagen appeared obviously in the lung tissues in the other groups. Data suggested that the contents of HYP increase in different degrees, furthermore the degree of PF aggravate gradually with the lapse of time. After the herbal compound treatment, HYP contents reduced, thus indirectly decreasing the contents of collagen, and acting on relieving the effect of PF. The results:⑴The PF rat model induced by BLM was successfully duplicated.⑵The herb compound of"strengthening earth to support metal and stasis-resolving"can reduce the HYP contents, so that it can improve the degree of fibrosis in PF rats.3 The study on the effects of"strengthening earth to support metal and stasis-resolving compound"on the expression of TGF-β1 and MCP-1 protein in PF rats.Through the method of immunohistochemistry, combining with image analysis, we observed the expression of TGF-β1 and MCP-1in lung tissues at different phases, further discussing the mechanism of PF from immunohistochemistry.The results showed as following: In the different time, the expression of TGF-β1 and MCP-1 protein in lung tissues are higher than the normal group. After"strengthening earth to support metal and stasis-resolving compound"treatment, the expression of TGF-β1 and MCP-1 protein all reduced in different degrees, which suggested that the herbal compound may be through inhibiting the high expression of TGF-β1 and MCP-1 protein, therefore making the degree of PF reduce.The results: After the herbal compound of"strengthening earth to support metal and stasis-resolving", the above makers may be improved effectively, so decreasing the degree of PF from immunohistochemistry.4 The study on the effect of the expression of TGF-β1mRNA in lung tissuesIn this study, using the method of RT-PCR and image analysis, we compare the expression of TGF-β1mRNA in lung tissues at different phases in every group, further discussing the mechanism of the herbal compound of"strengthening earth to support metal and stasis-resolving"from molecular biology.The results as following: TGF-β1mRNA expressed in lung tissues on different stages of PF, and there was significant difference in the expression intensity between the normal group and other groups. After"strengthening earth to support metal and stasis-resolving compound"treatment, the expression of TGF-β1mRNA in lung tissues was improved in different degrees, which suggested that the herbal compound may inhibit the expression of TGF-β1mRNA on PF rats.The results: After the herbal compound of"strengthening earth to support metal and stasis-resolving"treatment, the expression of TGF-β1mRNA obviously decreased, thus reducing the degree of PF from molecular biology.In summary, this study, by using BLM to induce PF model on rats, we apply the herbal compound of"strengthening earth to support metal and stasis-resolving"to treat the PF rats, thus discussing the effects of the herbal compound on PF rats. The results suggest that the herbal compound can effectively improve the abnormal status, reduce the content of HYP, suppress the high expression of TGF-β1mRNA, TGF-β1 and MCP-1 in lung tissues, thus relieving the degree of alveoli and fibrosis and blocking the pathological process, which provide the solid theoretical foundation for the clinical treatment in future.
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