醛糖还原酶抑制剂对糖尿病周围神经病变患者NLRP3炎性体表达的影响

Effect of aldose reductase inhibitor on the expression of NLRP3 inflammatory body in patients with diabetic peripheral neuropathy

目的探讨醛糖还原酶抑制剂对糖尿病周围神经病变(DPN)患者NLRP3炎性体表达的影响。方法回顾性分析2019年7月至2020年1月滨海县人民医院内分泌科收治的90例2型糖尿病(T2DM)患者的临床资料,其中糖尿病周围神经病变(DPN)组患者45例,非糖尿病周围神经病变(NDPN)组患者45例,同时选择在本院体检的健康人群45例作为对照组。采用Luminex200检测三组受检者血清白细胞介素(IL)-1β、IL-18浓度;采用实时定量PCR法(RT-PCR)检测人外周血单个核细胞(PBMCs)中NLRP3 mRNA、凋亡相关斑点样蛋白(ASC)mRNA、Caspase-1 mRNA表达水平;体外分离培养DPN患者PBMCs,并分三组[空白组(5 mmol/L葡萄糖)、高糖处理组(HG,25 mmol/L葡萄糖)、醛糖还原酶抑制剂非达司他(Fid)+HG组(Fid 10μmol/L+25 mmol/L葡萄糖)],通过RT-PCR法检测空白组、HG组及Fid+HG组NLRP3 mRNA、ASC mRNA、Caspase-1 mRNA,IL-1βmRNA、IL-18 mRNA水平。结果与对照组相比,DPN组及NDPN组患者的血清IL-1β、IL-18浓度均升高,且NDPN组升高更为明显,差异均有统计学意义(P<0.05);与对照组相比,NDPN组及DPN组患者PBMCs中NLRP3 mRNA、ASC mRNA、Caspase-1 mRNA表达升高,且NLRP3 mRNA表达在DPN组升高更为明显,差异均有统计学意义(P<0.05);DPN组患者PBMCs中NLRP3、ASC及Caspase-1表达与IL-1β(r=0.4051、0.3233、0.5430,P<0.01)、IL-18(r=0.3779、0.5022、0.3225,P<0.05)呈正相关;体外通过高糖刺激培养DPN患者的PBMCs,与空白组相比,HG组PBMCs中NLRP3 mRNA、ASC mRNA、Caspase-1 mRNA、IL-1βmRNA及IL-18 mRNA表达升高,差异均有统计学意义(P<0.05);与HG组相比,Fid+HG组PBMCs中NLRP3 mRNA、ASC mRNA、Caspase-1 mRNA、IL-1βmRNA及IL-18 mRNA表达降低,差异均有统计学意义(P<0.05)。结论醛糖还原酶抑制剂非达司他通过对DPN患者NLRP3炎性体的调控,可能有助于延缓该病的进程。

Objective To investigate the effect of aldose reductase inhibitor on the expression of NLRP3 in inflammatory bodies of patients with diabetic peripheral neuropathy(DPN).Methods A retrospective analysis was performed on 90 patients with type 2 diabetes mellitus(T2DM)diagnosed in the Department of Endocrinology,Binhai County People's Hospital from July 2019 to January 2020,including 45 patients with diabetic peripheral neuropathy(DPN)and 45 patients with non-diabetic peripheral neuropathy(NDPN),with 45 healthy people as the control group.The concentrations of serum interleukin(IL)-1βand IL-18 in three groups were measured by Luminex200.The expression of NLRP3 mRNA,apoptosis-related spot-like protein(ASC)mRNA,and Caspase-1 mRNA in human peripheral blood mononuclear cells(PBMCs)were detected by real-time quantitative PCR(RT-PCR).The PBMCs of DPN patients were isolated and cultured in vitro and divided into three groups:blank group(5 mmol/L glucose),high glucose treatment group(HG,25 mmol/L glucose),fidarestat(Fid)+HG group(Fid 10μmol/L+25 mmol/L glucose).The levels of NLRP3 mRNA,ASC mRNA,Caspase-1 mRNA,IL-1βmRNA,and IL-18 mRNA in each group were detected by RT-PCR.Results The levels of serum IL-1βand IL-18 were higher in DPN group and NDPN group than those of control group,especially higher in NDPN group.The expression of NLRP3 mRNA,ASC mRNA,and Caspase-1 mRNA in PBMCs of NDPN group and DPN group were higher than that in control group,and the expression of NLRP3 mRNA in DPN group was significantly higher than that in NDPN group.The expression of NLRP3,ASC,and Caspase-1 in PBMCs of DPN patients showed a positive correlation with the serum concentrations of IL-1β(r=0.4051,0.3233,0.5430,respectively,all P<0.01)and IL-18(r=0.3779,0.5022,0.3225,respectively,all P<0.05).The PBMCs of DPN patients were cultured by high glucose stimulation in vitro.Compared to the blank group,the expression of NLRP3 mRNA,ASC mRNA,Caspase-1 mRNA,IL-1βmRNA,and IL-18 mRNA in PBMCs of HG group were increased(P<0.05).Compared to HG group,the expression of NLRP3 mRNA,ASC mRNA,Caspase-1 mRNA,IL-1βmRNA,and IL-18 mRNA in PBMCs of Fid+HG group were decreased(P<0.05).Conclusion The aldose reductase inhibitor may delay the progression of the disease by regulating the NLRP3 inflammasome in DPN patients.