Effects of HRZ triple antituberculosis drug/TGF-β1 siRNA nanoliposomes on the expression of Ag85A and TGF-β1 in human BCG-infected human macrophages

English Keywords:Isoniazid/rifampicin/pyrazinamide(HRZ)  Triple antituberculosis drug/transforming growth factor-β1 siRNA nanoliposomes  Macrophage  Antituberculosis  Targeted gene silencing

English Abstract:

  【Abstract】 Objectives: To investigate the effects of isoniazid/rifampicin/pyrazinamide(HRZ) triple anti-tuberculosis/epidermal transforming growth factor(TGF)-β1 siRNA nanoliposomes on the expression of Ag85A and TGF-β1 in human macrophages in vitro. Methods: The human mononuclear cell strain THP-1 was induced to differentiate into macrophages in vitro. The experiment was divided into 4 groups. The blank group was cultured as macrophage alone. In the model group, macrophage and bacillus calmette-guerin(BCG) were cocultured at the ratio of 1∶5 for 4h to prepare a BCG-infected macrophage model. In the control group, BCG-infected macrophages and HRZ triple antituberculosis nanoliposomes were cocultured. In the experimental group, BCG-infected macrophages and HRZ triple anti-tuberculosis drug/TGF-β1 siRNA nanoliposome(C1 35mg/ml, C2 40mg/ml, and C3 50mg/ml) were cococultured. The mRNA expression levels of Ag85A and TGF-β1 in human macrophages were detected by RT-PCR. The protein expression levels of Ag85A and TGF-β1 were detected by western-blot method. Results: RT-PCR and western-blot results showed that the mRNA and protein levels of Ag85A and TGF-β1 were significantly up-regulated in the model group(BCG-infected macrophage model) compared with the blank group(macrophage culture alone)(P<0.05). The mRNA and protein expressions of Ag85A and TGF-β1 in the control group(cocultured with BCG-infected macrophages and HRZ triple anti-tuberculosis nano-liposomes) were significantly down-regulated compared with the model group(P<0.05); Compared with the model group and the control group, the mRNAs and protein expressions of Ag85A and TGF-β1 treated with HRZ triple antituberculosis drugs/TGF-β1 siRNA nanoliposomes in three different concentrations(C1 group, C2 group and C3 group) in the experimental group was further down-regulated(P<0.05). With the increase of nano-liposome concentration(35mg/ml, 40mg/ml, 50mg/ml), the expressions of TGF-β1 mRNA and protein decreased significantly(the difference between C1 group and C2 group and C3 group was statistically significant, P<0.05). Conclusions: The self-developed HRZ triple anti-tuberculosis drug/TGF-β1 siRNA nano-liposome has obvious down-regulation of Ag85A gene expression and TGF-β1 gene silencing.