| Objective:The purpose of this study was to investigate the effect of Wnt3a protein transfected with BMSCs(bone marrow mesenchymal stem cells)on inflammatory factors(IL-10;IL-6;IL-1β)of chronic obstructive pulmonary disease(COPD).Method:BMSCs were extracted and cultured by whole bone marrow adherent cell method.Rat bone marrow mesenchymal stem cells were transfected with Wnt3a lentivirus.The rat model of COPD was induced by two intratracheal instillations of LPS and exposed to cigarette smoke for 1 month.The experiment was divided into 4groups,and the healthy control group(Group C)was injected with 1ml PBS solution into normal rats via tail vein.COPD group(Group S):1ml PBS was injected into the body of COPD rats through tail vein.BMSCs group(group M):The labeled MSCs were injected into COPD rats by tail vein in the amount of 1×10~6/1MLPBS.Wnt3a-bmscs group(Group I):The P3 generation Wnt3a-BMscs were injected into COPD rats by tail vein,and the amount was 1×10~6/1MLPBS.Rats in each group were sacrificed on the 7th,14th and 28th day,and lung tissues were retained.Western blot was used to detect the expression of IL-10,IL-6 and IL-1βproteins in lung tissue,and QRT-PCR was used to detect the expression of IL-10,IL-6 and IL-1βmRNA in lung tissue.Result:1.The BMSCs extracted and cultured by flow cytometry showed high expression of CD29(96.3%)and CD44(69.2%),while low expression of CD34(0.25%)and CD45(0.74%).2.BMSCs were successfully transfected with Wnt3 lentivirus,and Wnt3a gene was highly expressed in COPD rats.3.Western blot analysis of IL-10,IL-6 and IL-1βprotein expression in lung tissue showed that:(1)The expression of pro-inflammatory factors IL-6 and IL-1βin COPD group increased at D7,D14 and D28 compared with the control group,showing an increasing trend,with the most obvious expression in D28(P<0.05).However,the expression of anti-inflammatory factor IL10 in COPD group at D7,D14 and D28 was decreased compared with that in control group,showing a decreasing trend,and the expression of anti-inflammatory factor IL10 was the most obvious in D28,P<0.05.(2)At the same time node on day 28,the protein expression levels of pro-inflammatory factors IL-6 and IL-1βin COPD group were higher than those in healthy control group,while the histone expression levels in BMSCs group and Wnt3a-BMSCs group were lower than those in COPD group,and the Wnt3a-BMSCs group was lower than that in BMSCs group,P<0.05.The protein expression of anti-inflammatory factor IL-10 in COPD group was lower than that in healthy control group,while the histone expression of BMSCs group and Wnt3a-BMSCs group was higher than that in COPD group,and Wnt3a-BMSCs group was higher than that in BMSCs group(all P<0.05).4.The expression levels of IL-10,IL-6 and IL-1βmRNA in lung tissues were detected by QRT-PCR:(1)Compared with the healthy control group,with time progression(D7;D14;D28),the expression of pro-inflammatory factors IL-6 and IL-1βshowed an increasing trend in COPD group,and D28 showed the most significant increase(all P<0.05).The expression of anti-inflammatory factor IL10 showed a decreasing trend in COPD group,and the decrease of D28 was the most significant(P<0.05).(2)At the same time point of 28 days,the pro-inflammatory factors IL-6 and IL-1βwere lower in the BMSCs group and Wnt3a-BMSCs group than in the COPD group(both P<0.001),and lower in the Wnt3a-BMSCs group than in the BMSCs group and Wnt3a-BMSCs group than in the BMSCs group(P<0.001).The anti-inflammatory factor IL-10 in the BMSCs group and Wnt3a-BMSCs group was higher than that in the COPD group,and the Wnt3a-BMSCs group was relatively higher than that in the BMSCs group and Wnt3a-BMSCs group(P<0.0001),the difference was statistically significant,suggesting that the anti-inflammatory effect of BMSCs was enhanced.Conclusion:Exogenous Wnt3a-BMSCs and BMSCs transplantation significantly promoted the expression of anti-inflammatory cytokine IL-10 in the lung tissues of COPD rats,while inhibited the expression of pro-inflammatory cytokines IL-6 and IL-1β,and the effect of Wnt3a-BMSCs on inflammatory cytokines was better than that of BMSCs alone. |
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