| ObjectiveWe observed the expression changes of buyanghuanwu decoction on HMGB1mRNA and TLR4mRNA in the rat model of pulmonary fibrosis that induced by bleomycin, to investigate the intervention effect of buyanghuanwu decoction on pulmonary fibrosis. and to provide evidence for Chinese medicine treatment of pulmonary fibrosis.Method144 SD male rats, weight 200±10g, after one week of adaptive feeding, rats were randomly divided into 6 groups:control group, model group, prednisone group, buyanghuanwu decoction low dose group, buyanghuanwu decoction middle dose group, buyanghuanwu decoction high dose group. In addition to the control group, the other 5 groups made pulmonary fibrosis model by using the classic method, which was established by a disposable injection 0.2-0.3ml bleomycin (5mg/kg) into its trachea and then quickly erected and rotated. Under the same conditions, the control group disposably injected the same amount of normal saline into its trachea. The rats were treated with medicines after 24 hours. The control group and model group were intervened by 10ml/kg·d distilled water, the prednisone group was given 4.2mg/kg·d prednisone solution, the low dose group was given buyanghuanwu decoction solution by 6.83g/kg·d, the middle dose group was given buyanghuanwu decoction solution by 13.66g/kg-d, the high dose group was given buyanghuanwu decoction solution by 27.32g/kg-d. During the 28 days of treatment, the general situation of rats was observed. At the 7th、14th and 28th day, the rats of six goups were killed by cutting the right femoral artery in batches. The rats’ chest were opened, their general situations were observed and their the right lung tissues were picked. HE staining and Masson staining were used to evaluated for alveolar inflammation and fibrosis in lung tissue, according to the Szapiel method is divided into 4 levels; RT-PCR method was used to detect the expression changes of HMGB1mRNA and TLR4mRNA in lung tissue.Results1. General situation:the rats of control group had smooth hair, strong body, normal food intake, sensitive reaction and increased body weight. The rats of model group were in low spirits, they had dry hair, trouble breathing, reduced food intake, and cyanosis of lips and limbs. The rats of each treatment group had a better spirit and stable breathing; their reduced activity, loss of appetite, cyanosis of lips and limbs and other symptoms had different degrees of improvement with the advance of time.2. Pathomorphology:The control group had no significant pathological changes. Compared with the control group, the model group and each treatment group showed inflammatory cell infiltration in the alveolar cavity, alveolar septum broadening, fibroblast and fibrocyte proliferation, and collagen fiber hyperplasia (P<0.05). Compared with the model group, the alveolar inflammation and fibrosis in each treatment group had varying degrees of relief (P<0.05). Compared with prednisone group, each dose group had no significant difference over the same period(.P>0.05). There was no significant difference between each dose group(P>0.05).3. The expression of HMGB1mRNA in lung tissue:At the 7th、14th and 28th day, the control group had a certain amount of basal expression. Compared with the control group, the other groups had different degrees of increase in the expression of HMGB1mRNA (P<0.05). Compared with the model group, each treatment group had different degrees of decrease at different time points (P<0.05). Compared with prednisone group, each dose group had no difference at the 7th day, but were significantly reduced at the 14th and 28th day (P<0.05).4. The expression of TLR4mRNA in lung tissue:Compared with the control group, the other groups had different degrees of increase in the expression of TLR4mRNA (P<0.05). Compared with the model group, each treatment group had different degrees of decrease at different time points (P<0.05). Compared with prednisone group, the expression of TLR4mRNA in each dose group was higher than that of prednisone group at the 7th day (P<0.05), but there was no significant difference at the 14th and 28th day.Conclusion1. At different time points, the pathological changes of pulmonary inflammation and fibrosis in the model group and treatment groups were higher than that in the control group, and the biological characteristics also had obvious changes. These changes were consistent with the references reported, and implied that we have successfully duplicated the rat model of pulmonary fibrosis.2. Each treatment group were down regulated HMGB1mRNA. Each dose group were significantly reduced at the 14th and 28th day, which illustrated that buyanghuanwu decoction mainly have acted on fibrosis stage and inhibited formation of late fibrosis.3. Each treatment group were down regulated TLR4mRNA, wherein the decline ranges of prednisone group were greater than each dose group at the 7th day, but there was no significant difference at the 14th and 28th day. It further illustrated that buyanghuanwu decoction mainly have acted on fibrosis stage. It also may be that TLR4 as a downstream signal of HMGB1, buyanghuanwu decoction down regulated HMGB1 to reduce expression of TLR4 and inhibited formation of late fibrosis. |
PDF Full Text Request