连续性血液净化对重型狼疮性肾炎合并急性肾损伤患者PYK2信号转导途径的影响

Effects of continuous blood purification on the PYK2 signaling pathway activity in severe lupus nephritis patients combined with acute renal injury

目的观察连续性血液净化(continuous blood purification,CBP)治疗对重型狼疮性肾炎合并急性肾损伤患者外周血单个核细胞富含脯氨酸的酪氨酸激酶2(proline-rich tyrosine kinase2,PYK2)信号通路活化的影响,探讨CBP治疗重型狼疮性肾炎合并急性肾损伤的作用机制。方法选取深圳市宝安区人民医院肾内科、风湿免疫科和重症医学科2012年1月~2016年4月收治的重型狼疮性肾炎合并急性肾损伤48例,将其随机分为观察组和对照组各24例,2组均给予常规治疗,观察组在常规治疗基础上给予CBP治疗,对照组在常规治疗基础上给予间歇性血液净化(intermittent hemodialysis,IHD)治疗。比较2组患者治疗前后系统性红斑狼疮疾病活动指数(systemic lupus erythematosus disease activity index,SLEDAI)、血清尿素氮(blood urea nitrogen,BUN)、肌酐(serum creatinine,SCr)水平及治疗有效率。Western-blotting测定治疗前、治疗后7天、14天、30天患者外周血单个核细胞磷酸化PYK2蛋白(phosphorylation-PYK2,p-PYK2)的表达,流式细胞术测定共刺激分子CD40L和CTLA4蛋白的表达。结果治疗30天后,与对照组相比,观察组患者SLEDAI(3.4±2.8比7.7±2.3,t=2.334,P=0.023)、BUN(6.4±3.8比16.7±5.3,t=3.930,P<0.001)、SCr(86.2±17.3比127.1±31.2,t=2.482,P=0.016)明显降低,差异有统计学意义;治疗后观察组有效率为91.7%,高于对照组的58.3%,差异有统计学意义(χ~2=4.552,P=0.013);治疗30天后,与对照组比较,观察组p-PYK2(0.732±0.206比1.110±0.152,t=3.196,P=0.007)、CD40L(9.4%比18.6%,χ~2=3.516,P=0.027)、CTLA4(7.2%比6.3%,χ~2=3.950,P=0.018)蛋白的表达明显降低,差异有统计学意义。结论 CBP对重型狼疮性肾炎合并急性肾损伤治疗效果明显,可以抑制患者外周血单个核细胞PYK2信号通路活化,重建机体免疫系统内稳状态,阻止肾脏病变的进一步发展,及时逆转肾功能。

Objective To investigate the effects of continuous blood purification（CBP） on the PYK2 signaling pathway activity in severe lupus nephritis patients combined with acute renal injury, and to explore the treatment mechanism of CBP. Methods A total of 48 severe lupus nephritis patients combined with acute renal injury admitted to our hospital from January 2012 to April 2016 were randomly divided into 2 groups： observation group（n=24） and control group（n=24）. In addition to the conventional therapy for both groups,CBP treatment was conducted for patients in observation group, and intermittent hemodialysis（IHD） treatment was conducted for those in control group. Systemic lupus erythematosus disease activity index（SLE-DAI）, serum urea nitrogen（BUN）, creatinine（SCr）, and effectiveness were compared between the two groups. p- PYK2, CD40 L and CTLA4 protein levels in peripheral blood mononuclear cells were assayed before and after the treatment. Results After the treatment for 30 days, SLEDAI, BUN and SCr were significantly lower in observation group than in control group（3.4±2.8 vs. 7.7±2.3, t=2.334, P=0.023 for SLEDAI;6.4±3.8 vs. 16.7±5.3, t=3.930, P0.001 for BUN; 86.2±17.3 vs. 127.1±31.2, t=2.482, P=0.016 for SCr）. The ratios of therapeutic effectiveness were 91.7% and 58.3% in observation group and control group respectively（χ~2=4.552, P=0.013）. After the treatment for 30 days, p-PYK2, CD40 L and CTLA4 levels were significantly lower in observation group than in control group（0.732 ± 0.206 vs. 1.110 ± 0.152, t=3.196, P=0.007 for pPYK2; 9.4% vs. 18.6%, χ~2=3.516, P=0.027 for CD40L; 7.2% vs. 16.3%, χ~2=3.950, P=0.018 for CTLA4）.Conclusion CBP can inhibit the activated PYK2 signaling pathway in severe lupus nephritis combined with acute renal injury, rebuild the homeostasis of immunological system, prevent kidney from further injuries, and thus improve the acute renal injury.