The Expression Of PDGF And HIF-1α In The Serum Of IPF Patients With Different Syndrome Types And The Effect Of Shenqi Cordycepes Recipe On The PI3K/AKT Signaling Pathway In Rats Induced By Bleomycin

Objective:1 To study the correlation between the levels of PDGF and HIF-1α in serum of IPF patients with deficiency of Qi and Yin and syndrome of stasis and internal heat of yin deficiency and syndrome types.2 To study the effect of Shenqi Cordyceps Recipe containing serum on bleomycin-induced PI3K/AKT signaling pathway in rats.Methods:1 Clinical study: Collect 60 patients with idiopathic pulmonary fibrosis.The TCM syndromes are Qi and Yin deficiency and stasis syndrome or Yin deficiency and internal heat syndrome,30 cases in each group.According to the syndrome type,they are divided into Qi and Yin deficiency and stasis syndrome group(Q),Yin deficiency and internal heat syndrome group(Y),30 healthy people served as normal control group(N).The serum levels of PDGF,HIF-1α,and expression in the three groups of cases were detected,and the levels of each index were compared between the groups according to the test results.2 In vitro cell experiments:(1)Select 20 SD healthy male rats and randomly divide them into 2 groups.Choose a one-time intratracheal instillation of bleomycin to create a model.The blank group uses normal saline instead of bleomycin for sham operation.After 28 days,the lung tissue was taken aseptically,and the primary lung microvascular endothelial cells were isolated and cultured.(2)Select 40 healthy male SD rats and randomly divide them into 2 groups,Shenqi Cordyceps Prescription Group(SQCC)and Blank Group(KB).SQCC group was gavage with Shenqi Cordyceps decoction every day,and KB group was gavage with the same amount of normal saline twice a day for 3days.One hour after the last gavage,blood was collected from the abdominal aorta and the serum was separated.(3)Immunofluorescence identification of rat lung microvascular endothelial cells and their purity.(4)Divide rat lung microvascular endothelial cells into seven groups,normal group(group A);model group(B);model group + 10% drug-containing serum group(C);model group + PI3K inhibitor group(D);Model group + 10% medicated serum + PI3K inhibitor group(E);model group +VEGF inhibitor(F);Model group + 10% medicated serum + VEGF inhibitor group(G).After corresponding treatments,observe the number of tubules formed in each group.(5)Divide rat lung microvascular endothelial cells into five groups: blank group(K),model group(MX),10% drug-containing serum group(SQ),PI3K inhibitor group(YZ),10%drug-containing serum Group + PI3K inhibitor group(SQYZ),using RT-q PCR technology to detect the relative expression of PI3KmRNA,HIF-1α mRNA,AKTmRNA and VEGFmRNA in each group.Result1 Clinical research1.1 Compared with group N,the content of PDGF in the serum of group Q and group Y was significantly increased(P<0.01);the PDGF content of patients in group Q was higher than that of group Y(P<0.05).1.2 Compared with the N group,the serum HIF-1α content of the Q group and the Y group were significantly increased(P<0.01);the HIF-1α content of the Q group was higher than the HIF-1α content of the Y group(P<0.05).2 In vitro cell experiments2.1 Immunofluorescence identification results: v WF immunofluorescence staining was positive;the cells were identified as P1 generation cells,and the purity of the cells reached more than 90%.2.2 Lumen formation results: Compared with group A,the number of tubules formed in group B increased significantly(P<0.01);compared with group B,the number of tubules formed in groups C,D,and F decreased(P<0.05);compared with group C,the number of tubules in group D was reduced There was no statistical difference in the number of formation(P>0.05);the number of tubule formation in groups E,G and C,D,and F was significantly reduced(P<0.05);compared with group E,the number in group G increased(P<0.05).2.3 Measurement results of HIF-1α mRNA,AKT mRNA,PI3K mRNA,and VEGF mRNA expression of rat lung microvascular endothelial cells in each group2.3.1 HIF-1αmRNA expression in lung microvascular endothelial cells of rats in each group: HIF-1αmRNA in the lung microvascular endothelial cells of rats in the K group has a certain basic expression;compared with the K group,the relative expression of HIF-1a mRNA in the MX group increased(P<0.01);Compared with MX,the relative expression of HIF-1a mRNA in SQ group,YZ group and SQYZ group decreased(P<0.05);compared with SQ group,there was no difference in the expression level of YZ group(P>0.05),The relative expression of HIF-1a mRNA in the SQYZ group decreased(P<0.05);compared with the YZ group,the relative expression of HIF-1a mRNA in the SQYZ group decreased(P<0.05).2.3.2 Expression of AKT mRNA in pulmonary microvascular endothelial cells of rats in each group: A certain amount of basal expression of AKT mRNA in pulmonary microvascular endothelial cells of rats in group K;compared with group K,the relative expression of AKT mRNA in group MX increased(P<0.01);Compared with MX group,SQ group,YZ group and SQYZ group,the relative expression of AKT mRNA decreased(P<0.05);compared with SQ group,the expression level of YZ group had no difference(P>0.05),SQYZ group was relatively AKT mRNA The expression level decreased(P<0.05);compared with the YZ group,the relative expression level of AKT mRNA in the SQYZ group decreased(P<0.05).2.3.3 Expression of PI3K mRNA in pulmonary microvascular endothelial cells of rats in each group: PI3K mRNA in the pulmonary microvascular endothelial cells of rats in group K had a certain basic expression;compared with group K,the relative expression of PI3K mRNA in group MX increased(P<0.01);Compared with the MX group,the relative expression of PI3K mRNA in the SQ,YZ and SQYZ groups decreased(P<0.05);compared with the SQ group,the expression in the YZ group increased(P<0.05),and the SQYZ group had a relatively higher expression of PI3K mRNA The expression level decreased(P<0.05);compared with the YZ group,the relative expression level of PI3K mRNA in the SQYZ group decreased(P<0.05).2.3.4 Expression of VEGF mRNA in rat lung microvascular endothelial cells: There was a certain amount of basic expression of VEGF mRNA in rat lung microvascular endothelial cells in group K;the relative expression of VEGF mRNA in MX group increased compared with group K(P<0.01);and Compared with the MX group,the relative expression of VEGF mRNA in the SQ,YZ and SQYZ groups decreased(P<0.05);compared with the SQ group,the expression in the YZ group increased(P<0.05),and the relative expression of VEGF mRNA in the SQYZ group decreased(P<0.05);Compared with the YZ group,the relative expression of VEGF mRNA in the SQYZ group decreased(P<0.05).Conclusion:1 In IPF patients,hypoxia leads to increased angiogenesis factors in the body,resulting in a hypercoagulable state of blood,which is manifested by the blood stasis syndrome of traditional Chinese medicine.2 Angiogenesis may be involved in the pathogenesis of IPF.Shenqi Cordyceps prescription may inhibit the angiogenesis of pulmonary microvascular endothelial cells through PI3K/AKT signaling pathway,thereby interfering with the formation of pulmonary fibrosis.3 Shenqichongcao Prescription may inhibit angiogenesis and interfere with the process of IPF through PI3K/AKT signaling pathway.