| Purpose:According to the theory of "different disease with the same treatment" of traditional Chinese medicine,Professor Shao Changrong’s experience in the field of silicosis was applied to treat idiopathic pulmonary fibrosis.Chinese medicine prescription Huaxian Decoction diriving from the effective experience of silicosis treating was used for the treatment of IPF.A randomized,controlled clinical study was conducted to evaluate the clinical efficacy of Huaxian Decoction in the treatment of IPF belong to deficiency of both qi and yin phlegm accumulating with stagnation type.Cell experiment in vitro was conducted to observe the effects of Huaxian Decoction lyophilized powder solution on proliferation,activation and secretion of extracellular matrix of human embryonic lung fibroblasts.The regulation of TNF-α cytokine and NF-κB signaling pathway was observed in order to explore the mechanism of Huaxian Decoction in the treatment of IPF.Methods:Clinical trials: A randomized,controlled clinical study was performed in 64 patients with IPF who met the inclusion criteria,33 patients in the treatment group,and 31 patients in the control group.Both groups were treated with N-acetylcysteine,while the treatment group was also treated with Chinese medicine Huaxian Decoction.The TCM syndrome score,lung function FVC,TLC,DLCO,6 minite walking distance,Borg dyspnea score,SGRQ quality of life score,serum cytokines TNF-α,TGF-β were compared before and after treatment.Cell experiment: TGF-β1 was used to induce proliferation and activation of human embryonic lung fibroblast MRC-5.Huaxian Decoction lyophilized powder solution was used to intervened.CCK method was used to detect the effect of Huaxian Decoction lyophilized powder solution on proliferation of MRC-5 cells.QRT-PCR method was used to detect the expression of α-SMA,type I collagen and type III collagen m RNA in order to observe the transformation of fibroblasts into myofibroblasts and extracellular matrix secretion.QRT-PCR method was used to detect the expression of TNF-α and NF-κB p65,IκBα and IKKβ m RNA,while Western Blot method was used to detect the protein expression of NF-κB P65 in order to observe the effect of Huaxian Decoction lyophilized powder solution on inflammatory response factor TNF-α and signaling pathway NF-κB.Result:1.TCM syndrome points: At the end of treatment,the main symptoms wheezing,cough,cough,chest tightness scores,secondary symptoms dry mouth thirst,spontaneous sweating,fatigue scores and TCM syndrome total score of the treatment group were decreased compared with before treatment.There were having significant difference between treatment before and after(P<0.01),The main symptom scores,secondary symptom scores and TCM syndrome total score of the control group were increased compared with those before treatment.The cough score had significantly difference from before treatment(P<0.05),and the rest symptom scores and TCM syndrome scores had no significant difference from before treatment.There had significant difference between the two groups after treatment of the main symptom scores,secondary symptom scores and TCM syndrome total score(P<0.01).2.The efficacy of TCM syndromes: The effective rate of the treatment group was 54.5%,while the effective rate of the control group was 3.2%.The effective rate of the treatment group was higher than that of the control group through the rank sum test.3.Pulmonary function: After treatment,the main index of lung ventilation function FVC increased(88±168)m L from baseline in the treatment group,while decreased(198±195)m L in the control group.There was no significant difference in FVC of two groups compared with treatment before separately.The difference of FVC between the two groups was having statistically significant after treatment(P<0.05).After the treatment,there was not significantly difference in TLC of two groups compared with treatment before separately.There was significant difference in TLC between the two groups after treatment(P<0.05).after treatment,There was no significant difference in DLCO of two groups compared with treatment before.There was significant difference in DLCO between the two groups after treatment(P<0.05).4.Six-minute walk test: After treatment,the treatment group had a significant increase in 6MWD compared with treatment before(P(27)0.05),while the control group had a decrease in 6MWD compared with treatment before,but there was no significant difference.There had significant difference between the two groups after treatment(P(27)0.01).5.Borg dyspnea score: After treatment,the Borg score of the treatment group was significantly lower than treatment before(P(27)0.01),the Borg score of the control group was higher than treatment before,but there was no significant difference between treatment before and after.There had significant difference between two groups after treatment(P(27)0.01).6.SGRQ quality of life score: After treatment,the symptom part,activity part,affected part and total score of the treatment group were lower than those before treatment,while these scores were increased in the treatment group after treatment.There was no significant difference between treatment before and after of the two group.There had significant difference in these scores between the two groups after treatment(P(27)0.05),and the scores of the treatment group were significantly lower than the control group.7.Serum cytokines: After treatment,TNF-α and TGF-β in serum of the treatment group were decreased compared with treatment before,and they were having significant statistics difference(P(27)0.05).The TNF-α and TGF-β of the control group were were increased compared with treatment before,but they were not having significant difference.There was significant difference between the two groups after treatment(P(27)0.05),and the treatment group was lower than the control group.8.The effect of Huaxian Decoction lyophilized powder solution on the proliferation of MRC-5 cells: The cell proliferation level was detected by CCK method.Compared with the blank control group,the cell proliferation level of the model control group was significantly increased(P<0.01).Compared with the model control group,the proliferation level of MRC-5 cells in the medium and high dose groups of Huaxian Decoction lyophilized powder solution decreased significantly(P<0.01).9.The effect of Huaxian Decoction lyophilized powder solution on the activation of MRC-5 cells: QRT-PCR was used to detect the level of α-SMA,type I collagen and type III collagen m RNA.Compared with the blank control group,the expression of α-SMA,type I collagen and Type III collagen m RNA in the model group were significantly increased(P < 0.01).Compared with the model group,the expression of α-SMA,type I collagen and type III collagen m RNA in Huaxian Decoction lyophilized powder solution groups were significantly sigdecreased(P<0.05 or P<0.01).10.The effect of Huaxian Decoction lyophilized powder solution on inflammatory factor TNF-α and inflammatory signaling pathway NF-κB:QRT-PCR was used to detect the level of TNF-α,NF-κB p65,IκBα and IKKβ m RNA.The results showed that compared with the blank control group,The expression of TNF-α and NF-κB p65 m RNA in the model group were significantly increased(P<0.05 or P<0.01).The expression of IκBα and IKKβ m RNA were significantly decreased(P<0.01).Compared with the model group,the expression of TNF-α and NF-κB p65 m RNA of each Huaxian Decoction lyophilized powder solution group were decreased,the expression of IκBα,IKKβ m RNA of each Huaxian Decoction lyophilized powder solution group were increased(P<0.05,P<0.01 or P>0.05).The protein expression of NF-κB P65 was detected by Western Blot.The results showed that the protein expression of NF-κB P65 in the model group were significantly increased compared with the blank control group(P<0.01).Compared with the model group,the protein expression of NF-κB P65 of each Huaxian Decoction lyophilized powder solution group were decreased,they were having statistically different(P<0.01).Compared with the model group,the expression level of NF-κB P65 protein in each chemical fiber freeze-dried powder solution group decreased,which was statistically significant(P<0.01).Conclusion:1.Chinese medicine Huaxian Decoction could delay the decline of lung function FVC,TLC,DLCO in patients with IPF,improve symptoms,increase exercise tolerance and elevate quality of life.2.Chinese medicine Huaxian Decoction could decrease serum cytokines TNF-α and TGF-β of IPF patients.It was speculated that Huaxian Decoction may exert a therapeutic effect on IPF patients by inhibiting the inflammatory reaction.3.Chinese medicine Huaxian Decoction lyophilized powder solution could inhibit the proliferation of human embryonic lung fibroblasts,reduce the expression of α-SMA,type I collagen and type III collagen m RNA,reduce the extracellular matrix secretion.This indicated that Huaxian Decoction has anti-fibrotic effects in vitro cell experiments.4.Chinese medicine Huaxian Decoction lyophilized powder solution played a therapeutic role in the progression of IPF disease by inhibiting the secretion of inflammatory factor TNF-α and the activation of NF-κB inflammatory signaling pathway. |
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