蛇床子素调节Mincle/Syk/NF-κB信号通路对肾病综合征大鼠的治疗作用

Therapeutic Effects of Osthol in Modulating Mincle/Syk/NF-κB Signaling Pathway in Rats with Nephrotic Syndrome

目的:探讨蛇床子素(OST)调节巨噬细胞诱导性C型凝集素样受体(Mincle)/脾脏酪氨酸激酶(Syk)/核因子κB(NF-κB)信号通路对肾病综合征(NS)大鼠的治疗作用。方法:随机选择10只大鼠记为N组,其它大鼠构建NS模型,将建模成功的50只大鼠随机平分为NS组(模型组)、L-OST组(10mg/kg OST)、M-OST组(20mg/kg OST)、H-OST组(40mg/kg OST)、H-OST+TDB组(40mg/kg OST+50μg TDB/周),OST每天注射1次,连续治疗4周。ELISA法检测血清中炎性因子(IL-1β、IL-6、TNF-α)、氧化应激指标(SOD、MDA、LDH)、24h尿液中UP水平;通过全自动化学分析仪检测BUN、Scr水平;HE染色以及Masson染色检测肾脏病理变化;TUNEL染色检测肾脏细胞凋亡;Western blot检测collagen I、Ly6g、Ki-67、Mincle/Syk/NF-κB通路蛋白表达。结果:N组肾组织结构正常,染色清晰,NS组出现大量间质炎性细胞浸润、肾小管萎缩、大量空泡的现象,NS组较N组24h UP、Scr、BUN含量、IL-1β、IL-6、TNF-α、MDA、LDH水平、胶原容积分数、凋亡率、collagen I、Ly6g、Ki-67、Mincle、Syk、p-NF-κB/NF-κB蛋白水平均显著升高(P<0.05),SOD水平显著下降(P<0.05);与NS组相比,L-OST组、M-OST组、H-OST组均改善炎性细胞浸润、肾小管萎缩现象,24h UP、Scr、BUN含量、IL-1β、IL-6、TNF-α、MDA、LDH水平、胶原容积分数、凋亡率、collagen I、Ly6g、Ki-67、Mincle、Syk、p-NF-κB/NF-κB蛋白水平均显著下降(P<0.05),SOD水平显著升高(P<0.05),且H-OST组治疗效果最佳;TDB消除了OST对NS大鼠肾损伤的改善作用。结论:OST可能通过抑制Mincle/Syk/NF-κB信号通路减轻OST大鼠的肾损伤。

Objective:To investigate the therapeutic effect of osthol(OST)on nephrotic syndrome(NS)in rats by regulating macrophage-inducible C-type lectin(Mincle)/spleen tyrosine kinase(Syk)/nuclear factor-κB(NF-κB)signal pathway.Methods:Ten rats were randomly selected and recorded as group N,and other rats were used to construct NS model.The 50 successfully modeled rats were randomly divided into NS group(model group),L-OST group(10 mg/kg OST),M-OST group(20 mg/kg OST),H-OST group(40 mg/kg OST),H-OST+TDB group(40 mg/kg OST+50μg TDB/week).OST was injected once a day for 4 consecutive weeks.The levels of inflammatory factors(IL-1β,IL-6,TNF-α),oxidative stress indicators(SOD,MDA,LDH)in serum and UP in 24h urine were detected by ELISA;the levels of BUN and Scr was measured by automatic chemical analyzer;HE staining and Masson staining were used to detect the pathological changes of kidney;TUNEL staining was used to detect renal cell apoptosis;Western blot was used to detect the expression of collagen I,Ly6g,Ki-67,and Mincle/Syk/NF-κB pathway proteins.Results:In group N,the renal tissue structure was normal and the staining was clear.In NS group,a large number of interstitial inflammatory cells infiltrated,renal tubules atrophied,and a large number of vacuoles were observed,compared with group N,the contents of 24h UP,Scr,BUN,levels of IL-1β,IL-6,TNF-α,MDA,LDH,the collagen volume fraction,apoptosis rate,the levels of collagen I,Ly6g,Ki-67,Mincle,Syk,p-NF-κB/NF-κB proteins in NS group increased significantly(P<0.05),the level of SOD decreased significantly(P<0.05);compared with NS group,the inflammatory cell infiltration and renal tubule atrophy in the L-OST group,M-OST group and H-OST group were improved,the contents of 24h UP,Scr,BUN,levels of IL-1β,IL-6,TNF-α,MDA,LDH,the collagen volume fraction,apoptosis rate,the levels of collagen I,Ly6g,Ki-67,Mincle,Syk,p-NF-κB/NF-κB proteins decreased significantly(P<0.05),the level of SOD increased significantly(P<0.05),the treatment effect of H-OST group was the best;TDB eliminated the ameliorative effect of OST on renal injury in NS rats.Conclusion:OST may alleviate renal injury in OST rats by inhibiting the Mincle/Syk/NF-κB signal pathway.