摘要
目的 探讨大黄蛰虫丸调节IL-1β/NF-κB/NLRP3信号通路对动脉粥样硬化(AS)大鼠主动脉炎性损伤的影响。方法 将40只大鼠分为正常对照组(NS)、模型组、地奥心血康干预组(阳性组)及大黄蛰虫丸干预组(大黄蛰虫丸组),每组各10只。除正常组外,余下3组小鼠均构建AS模型。应用油红染色对主动脉病理切片情况进行观察,采用EILSA法检测血清脂质水平[甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)]、血清炎症指标(IL-6、IL-1β、TNF-α水平);应用流式细胞术对外周血Th17/Treg细胞水平进行检测,并通过RT-qPCR法和Western blot分别检测IL-1β/NF-κB/NLRP3信号通路mRNA和蛋白表达水平,并计算AI指数。结果 除NC组大鼠以外,其他各组大鼠动脉组织中均出现内膜增厚、斑块沉积和不连续、大量炎症细胞浸润等现象;其中模型组动脉组织损伤现象最为显著,阳性组和大黄蛰虫丸组较模型组明显改善,动脉组织结构情况与NC组相近。模型组血清TC[(9.82±1.46)mmol/L]、TG[(9.82±1.46)mmol/L]、LDL-C[(2.93±0.22)mmol/L]、IL-6[(97.65±9.11)ng/L]、TNF-α[(93.21±11.17)ng/L]水平及外周血Th17[(3.56±0.34)%]、Treg细胞[(4.41±0.38)%]水平高于NC组[分别为(3.71±0.57)mmol/L、(6.47±0.92)mmol/L、(0.94±0.15)mmol/L、(16.52±2.76)ng/L、(5.45±0.84)ng/L、(1.83±0.16)%、(7.72±0.73)%](均P<0.05),HDL-C水平[(2.92±0.31)mmol/L]低于NC组[(0.95±0.08)mmol/L](P<0.05);与模型组相比,大黄蜇虫丸组和阳性组血清TC[分别为(3.92±0.72)mmol/L、(3.71±0.65)mmol/L]、TG[分别为(6.71±0.83)mmol/L、(6.83±0.72)mmol/L]、LDL-C[分别为(1.15±0.11)mmol/L、(1.11±0.13)mmol/L]、IL-6[分别为(19.83±2.55)ng/L、(18.51±3.72)ng/L]、TNF-α[分别为(17.08±1.72)ng/L、(15.92±1.56)ng/L]水平及外周血Th17[分别为(2.11±0.25)%、(2.06±0.22)%]、Treg细胞[分别为(7.45±0.76)%、(7.31±0.72)%]水平偏低,HDL-C偏高[分别为(2.18±0.72)mmol/L、(2.23±0.61)mmol/L],趋近于NC组(均P<0.05)。与NC组AI指数(0.35±0.08)比较,模型组、阳性组和大黄蛰虫丸组(分别为9.71±2.04、1.21±0.15、0.83±0.17)均明显升高(均P<0.05),其中阳性组和大黄蛰虫丸组AI指数低于模型组(均P<0.05)。模型组大鼠的IL-6、TNF-α水平和外周血IL-1β、NF-κB、NLRP3 mRNA和信号通路蛋白表达水平均较NC组、阳性组和大黄蛰虫组明显偏高(均P<0.05)。结论 大黄蛰虫丸对动脉粥样硬化大鼠主动脉的炎性损伤可能具有明显的改善作用,可能是通过抑制IL-1β/NF-κB/NLRP3信号通路来实现治疗效果。
Objective To explore the effect of Dahuang zhechong pill on the inflammatory injury of aorta in atherosclerotic(AS)rats by regulating IL-1β/NF-κB/NLRP3 signaling pathway.Methods Forty rats were divided into normal control group(NS),model group,Di'ao Xinxuekang intervention group(positive group)and Dahuang zhechong pill intervention group(Dahuang zhechong pill group),with 10 rats in each group.Except for the normal group,the other three groups of mice all built AS models.The pathological sections of aorta were observed by oil red staining,and serum lipid levels[triglyceride(TG),high density lipoprotein cholesterol(HDL-C),total cholesterol(TC)and low density lipoprotein cholesterol(LDL-C)],serum inflammatory indexes[interleukin(IL)-6,IL-1βand tumor necrosis factor(TNF)]were detected by EILSA method.The peripheral blood Th17/Treg cell level was detected by flow cytometry,and the mRNA and protein expression levels of IL-1β/NF-κB/NLRP3 signaling pathway were detected by RT-qPCR and Western blot,respectively,and the AI index was calculated.Results Intima thick-ening,plaque deposition,discontinuity and infiltration of a large number of inflammatory cells were found in arterial tissues of rats in other groups except NC group.Among them,the injury of arterial tissue in the model group is the most obvious,and the positive group and Dahuang zhechong pill group are obviously improved compared with the model group,and the arterial tissue structure is similar to that in the NC group.The levels of serum TC(9.82 mmol/L±1.46 mmol/L),TG(9.82 mmol/L±1.46 mmol/L),LDL-C(2.93 mmol/L±0.22 mmol/L),IL-6(97.65 ng/L±9.11 ng/L),TNF-α(93.21 ng/L±11.17 ng/L)and peripheral blood Th17(3.56±0.34)%and Treg cells(4.41±0.38)%in the mod-el group were higher than those in the NC group(3.71 mmol/L±0.57 mmol/L),(6.47 mmol/L±0.92 mmol/L),(0.94 mmol/L±0.15 mmol/L),(16.52 ng/L±2.76 ng/L),(5.45 ng/L±0.84 ng/L),(1.83%±0.16%),(7.72%±0.73%),re-spectively]and the level of HDL-C(2.92 mmol/L±0.31 mmol/L)was lower(0.95 mmol/L±0.08 mmol/L)(all P<0.05).Compared with the model group,in Dahuang zhechong pill group and positive group,the levels of serum TC(3.92 mmol/L±0.72 mmol/L,3.71 mmol/L±0.65 mmol/L),TG(6.71 mmol/L±0.83 mmol/L,6.83 mmol/L±0.72 mmol/L),LDL-C(1.15 mmol/L±0.11 mmol/L,1.11 mmol/L±0.13 mmol/L,IL-6(19.83 ng/L±2.55 ng/L,18.51 ng/L±3.72 ng/L)and TNF-α(17.08 ng/L±1.72 ng/L,15.92 ng/L±1.56 ng/L),and the levels of peripheral blood Th17(2.11%±0.25%),2.06%±0.22%)and Treg cells(7.45%±0.76%),(7.31%±0.72%)were lower,and HDL-C(2.18 mmol/L±0.72 mmol/L,2.23 mmol/L±0.61 mmol/L)was higher,which was close to that in NC group(all P<0.05).Compared with NC group,the AI index of model group,positive group and Dahuang zhechong pill group increased significantly NC group(0.35±0.08,9.71±2.04,1.21±0.15 and 0.83±0.17,respectively)(all P<0.05),and the AI index of posi-tive group and Dahuangzhechong pill group was lower than that of model group(all P<0.05).The expression levels of IL-1β,NF-κB,NLRP3 mRNA and signal pathway protein in peripheral blood of rats in model group were signifi-cantly higher than those in NC group,positive group and rhubarb sting group(all P<0.05).Conclusion Dahuang zhechong pill can obviously improve the inflammatory injury of aorta in atherosclerotic rats,which may be achieved by inhibiting IL-1β/NF-κB/NLRP3 signaling pathway.
作者
汪云鑫
苏乙花
顾民华
WANG Yun-xin;SU Yi-hua;GU Min-hua(Cardiology Department of Hainan Provincial Hospital of Traditional Chinese Medicine,Haikou,Hainan Province 570000,China;Cancer Interventional Department of Hainan Provincial Hospital of Traditional Chinese Medicine,Haikou,Hainan Province 570000,China)
出处
《解剖学研究》
CAS
2024年第4期315-321,共7页
Anatomy Research
基金
国家自然科学基金项目(82160899)。
