| Objective: To observe the effect of Yiqiyangyinquyu recipeon bleomycine induced idiopathic pulmonary firosis in ratsrelated cytokines, approach its functional mechanism, andexplore the possible mechanisms of IPF from the view of TCM,which will provide practical and theoretical evidence fordeveloping new method of treating and preventing IPF.Methods: 120 healthy SD rats (paroecious about half) raisedin the experimental environment about one week were randomlydivided into five groups (24 per-group):①Control group:pyrogen-free normal saline (NS, 200μl per rat) was instilledintratracheally;②model group,③Prednisone group,④low-dosegroup and⑤high-dose group: bleomycin A5 was instilledintratracheally (5mg·kg-1).From the second day after instillation, control group andmodel group were filled with NS (10ml/kg),drenched byisotonic Na chloride; low-dose group and high-dose group weredrenched with Chinese crude drug elixation differently (12.3g·kg-1and 24.6g·kg-1, 7 and 14 times the dosage of the adult'sclinical drug consumption); prednisone group were drenchedwith prednisone(6mg·kg-1). The rats in each group were decapitated and sampled bloodon the 7th,14th,28th day after making models, immediatelycentrifuge 5 min with 4℃3500rpm, separation serum, and ofwhich the lung tissue were taken to make into 10％homogenate.In the lab, we measured the level ofTNF-α,TGF-β1,IFN-γ,HGF in both serum and lung tissue. Taken lung tissue about 1 cm×1cm×1cm, which were fixed with 4％formaldehyde,embedded with paraffin, maken slice by HE and masson stain.And observe the change of lung tissue's pathomorphology, andthe expression of TGF-β1 on the lung was detected byimmunohistochemistry.Results:1 Observation of the common condition:Control group ate and drunk normally, reacted sensitively,the fur was luster, the urine and stool were normal, with nodeath. Model group scrunched, coughed and breathed difficultly,had listlessness and withered fur. High-dose group and low-dosegroup gradually had the better mind,fur and appetite, breathedsmoothly, and their activity increased; prednisone groupalsohad better condition, but inferior to high-dose group andlow-dose group.2 The promotion fibrosis cytokines TNF-α, TGF-β1 contentschange in serum and lung tissuea of each group rats2.1 The TNF-αcontents change in serum and lung tissue of eachgroup ratsThe results were as follows: Compared with the same time points of control group, the TNF-αcontents in serum and lungtissue of model group were increased obviously (P＜0.01).Compared with the same time points of model group, the TNF-αcontents in serum and lung tissue of the treatment groups weredecreased obviously (P＜0.05orP＜0.01). Compared withprednisone group, the TNF-αcontents in serum of high-dosegroup were decreased on the 7th and 28th day (P＜0.05orP＜0.01);The TNF-αcontents in lung tissue were decreased on the sametime points (P＜0.05orP＜0.01).2.2 The TGF-β1 contents change in serum and lung tissue ofeach group ratsThe results were as follows: Compared with the same timepoints of control group, the TGF-β1 contents in serum and lungtissue of model group were increased obviously (P＜0.01).Compared with model group, the TGF-β1 contents in serum ofprednisone group were decreased on the 7th day (P＜0.01); theTGF-β1 contents in serum of Chinese medicinal formulae groupson the sane time points (P＜0.01). Compared with the sametime points of model group, the TGF-β1 contents in lung tissuewere decreased (P＜0.01). Compared with the same time pointsof prednisone group, the TGF-β1 contents in serum and lungtissue of high-dose group were decreased (P＜0.05orP＜0.01).3 The resistance fibrosis cytokines IFN-γ, HGF contents changein serum and lung tissue of each group rats3.1 The IFN-γcontents change in serum and lung tissue of eachgroup rats The results were as follows: Compared with control group,the IFN-γcontents in serum and lung tissue were increased(P＜0.01)on the 7th day, after a steady decline, the IFN-γcontents inserum and lung tissue were decreased obviously on the 28th day(P＜0.05orP＜0.01). Compared with model group, the IFN-γcontents in serum and lung tissue of prednisine group weredecreased on the 7th day (P＜0.01), the IFN-γcontents in serumand lung tissue of Chinese medicinal formulae groups weredecreased on the 7th day(P＜0.01) and increased on the 14thand 28th day (P＜0.05orP＜0.01). Compared with prednisonegroup, the IFN-γcontents in serum and lung tissue of Chinesemedicinal formulae groups were decreased on the 7th day (P＜0.01) and increased on the 14th and 28th day (P＜0.05orP＜0.01).Compared with low-dose group, the IFN-γcontents in serum ofhigh-dose group were decreased on the 7th day (P＜0.01), andthe IFN-γcontents in lung tissue were increased on the 14th and28th day (P＜0.05).3.2 The HGF contents change in serum and lung tissue of eachgroup ratsThe results were as follows: The HGF contents in serum ofmodel group were increased at peak value on the 7th day,although after a steady decline, they were still higher thancontrol group (P＜0.05orP＜0.01). Compared with the same timepoints of model group, the HGF contents in serum of Chinesemedicinal formulae groups were decreased (P＜0.05orP＜0.01).Compared with control group, the HGF contents in lung tissue were increased (P＜0.01) on the 7th day, after a steady decline,the HGF contents in lung tissue were decreased obviously onthe 28th day (P＜0.01). Compared with the same time points ofmodel group, the HGF contents in lung tissue of Chinesemedicinal formulae groups were increased (P＜0.05orP＜0.01).Compare with prednisone group, the HGF contents in bloodserum of low-dose group were decreased on the 14th day(P＜0.05), the HGF contents in blood serum of high-dose groupwere decreased on the 14th and 28th day (P＜0.05); The HGFcontents in lung tissue of Chinese medicinal formulae groupswere increased on the 14th and 28th day (P＜0.05).4 Observation of the lung's histomorphology4.1 Macroscopic observation: The lung's appearance of controlgroup was normal with color pink, surface smooth, elasticitygood and lobes of lung edge sharpness. Of model group, lungson the 7th day were bigger and spotty hemorrhage andecchymosis were seen; lungs on the 14th day were gray, onwhich there were inequality of size nodus, sppotty hemorrhage;on the 28th day, lungs were gray, smaller and harder, with nodusand bead-like aerate vacuole on the surface. The lung'sappearance of each treatment group was better than that ofmodel group.4.2 Obeservation under light microscope: Severe injuries wereobserved in lung tissues of model group, which werecharacterized by widened septa of alveoli, diffuse infiltration ofinflammatory cells accompanied by atrophied or disappeared alveolitis, and compensatory alveolar emphysema. In the lateperiod, midrange or severe pulmonary interstitial fibrosisdeveloped (More collagenous fibers in lung matrix wereobserved by Masson dyeing). Compared with model group,alveolitis and pulmonary fibrosis of each treatment group weredeclined significantly, especially in high-dose group.5 The expression of TGF-β1 in lungs of each group ratsThe results were as follows: The control group have poorexpressions in the lung tissue; compared with the same timepoints of control group, the degree of TGF-β1's expression wereenhanced in model group; compared with model group at thesame time points, the expression were decreased in the threetreatments (P＜0.01). There was no statistical significanceamong three treatment groups (P＞0.05).Conclusion:1 Under the condition of IPF, the promotion fibrosis cytokinesTNF-α,TGF-β1 contents in serum and lung tissue are increasedand the resistance fibrosis cytokines IFN-γ,HGF contents aredecreased, tiny epithetial tissues are continually insulted andpara-renovation after insulted by these changes. This alleosisprobably causes the pathogenesis of pulmonary fibrosis.2 Yiqiyangyinquyu recipe brings down the TNF-αand TGF-β1contents in serum and lung tissue, dajusts the IFN-γand HGFcontents, thus attenuates the promation of pulmonary fibrosisformation by cytokine network, inhibits the fibrosis evolution,gain the ends that preventing and curing pulmonary fibrosis. 3 Under the condition of IPF, lung tissue having hypsoexpression character of TGF-β1, indicates TGF-β1 promotes thefibrosis formation; Yiqiyangyinqu recipe relieves the degree ofPF through down regulation to the hypso-expressed TGF-β1inhibiting the deposition of ECM.
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