Effects of Xinkang Granules-contained Serum on the Proliferation of Cardiac Fibroblasts via Regulating GAS5-miR21
10.19879/j.cnki.1005-5304.202306159
- VernacularTitle:心康冲剂含药血清调控GAS5-miR21对心肌成纤维细胞增殖的影响
- Author:
Rongfang LIU
1
;
Bing GUO
;
Yi YU
;
Yuan LI
;
Min WANG
Author Information
1. 暨南大学附属江门市五邑中医院,广东 江门 529000
- Keywords:
cardiac fibrosis;
cell cycle;
Xinkang Granules;
GAS5;
miR-21;
cardiac fibroblasts
- From:
Chinese Journal of Information on Traditional Chinese Medicine
2024;31(9):102-109
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the effects of Xinkang Granules-contained serum on the proliferation of cardiac fibroblasts(CFs)induced by angiotensin Ⅱ(Ang Ⅱ);To explore its mechanism of inhibiting the proliferation of CFs.Methods Taking out the heart of SD suckling rats,extracting the CFs from the hearts by differential attachment,and then they were used to replicate cell proliferation model induced by Ang Ⅱ.After transfection with miR-21 overexpressing lentivirus and growth arrest specific transcript 5(GAS5)silencing lentivirus,Xinkang Granules-contained serum was intervened for 24 hours,respectively.Cell viability was detected by cell CCK-8 method,cell cycle was detected by flow cytometry,α-smooth muscle actin(α-SMA)was detected by immunofluorescence,RT-qPCR was used to detect the mRNA expression of GAS5,miR-2l,phosphatase and tensin homolog(PTEN),Western blot was used to detect the expressions of PTEN,protein kinase B(Akt),Cyclin D1 and cyclin-dependent kinase 4(CDK4)protein.Results Compared with the blank group,the activity of CFs in the model group increased,the proportion of G0/G1 phase cells decreased,the proportion of S phase cells increased,the positive expression of α-SMA increased,the expressions of GAS5,PTEN mRNA and PTEN protein decreased,the expressions of miR-21 mRNA and Akt,Cyclin D1,CDK4 protein increased,with statistical significance(P<0.05).Compared with the model group,the activity of CFs in the Xinkang Granules group decreased,the proportion of G0/G1 phase cells increased,the proportion of S phase cells decreased,the positive expression of α-SMA decreased,the expressions of GAS5,PTEN mRNA and PTEN protein increased,while the expressions of miR-21 mRNA and Akt,Cyclin D1 and CDK4 protein decreased;The activity of CFs increased in the miR-21 overexpression group and the GAS5 silencing group,and the proportion of G0/G1 phase cells decreased,the proportion of S phase cells increased,and the positive expression of α-SMA increased,the expressions of GAS5,PTEN mRNA and PTEN protein decreased,while the expressions of miR-21 mRNA and Akt,Cyclin D1 and CDK4 protein increased,with statistical significance(P<0.05).Compared with the miR-21 overexpression group and the GAS5 silencing group,the miR-21 overexpression+Xinkang Granules group and the GAS5 silencing+Xinkang Granules group showed a decrease in CFs activity and an increase in the proportion of G0/G1 phase cells,the proportion of S phase cells decreased,the positive expression of α-SMA decreased,the expressions of GAS5,PTEN mRNA and PTEN protein increased,and the expressions of miR-21 mRNA and Akt,Cyclin D1,CDK4 protein decreased,with statistical significance(P<0.05).Conclusion Xinkang Granules-contained serum can inhibit the increase of miR-21 and Akt expression caused by silence GAS5,resist the decrease of GAS5 and PTEN caused by overexpress miR-21,inhibit the proliferation cycle of CFs,promote GAS5 to play the role of"molecular sponge",and improve the excessive proliferation of CFs induced by Ang Ⅱ.
