|摘要||目的 探讨NADPH氧化酶系统(NOXs)对单侧输尿管梗阻(UUO)大鼠肾间质纤维化的影响.方法 雄性Wistar大鼠按随机区间法分为假手术组(Sham,n=8)、假手术+夹竹桃麻素(apocynin)治疗组(SA,n=8)、UUO组(n=8)和UUO+夹竹桃麻素治疗组(UA,n=8).SA及UA组自手术日起每天给予夹竹桃麻素100 mg/kg灌胃治疗,Sham组和UUO组给予等量清水灌胃.术后第7天处死取材,采用ELISA及Western印迹法分别检测肾脏组织超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性,8-异构前列腺素F2α(8-iso-PGF2α)浓度,NOX2、NOX4及α平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(COL-Ⅰ)的表达,ERK1/2激酶的活化.结果 与Sham组相比,UUO大鼠第7天NOX2、NOX4表达及8-iso-PGF2α水平显著升高(P＜0.05),同时伴有肾组织COL-Ⅰ和α-SMA表达升高、ERK1/2激酶活化增加(P＜0.05).夹竹桃麻素治疗显著降低NOX2、NOX4的表达及8-iso-PGF2α水平(P＜0.05),同时大鼠肾组织α-SMA及COL-Ⅰ表达显著下降、ERK1/2活化降低(P＜0.05).夹竹桃麻素治疗对肾组织SOD及CAT活性无明显影响.结论 NADPH氧化酶系统通过促氧化应激参与梗阻性肾病大鼠肾间质纤维化的进展,抑制该系统可能作为抑制肾间质纤维化的治疗靶点.
Objective To clarify whether the NADPH oxidases (NOXs) family contributed to the reactive oxygen species (ROS) production and subsequent interstitial fibrosis in unilateral ureter obstruction (UUO) rats.Methods Male Wistar rats were randomly divided into sham operation group (n =8),sham operation + apocynin treatment group (n =8),UUO operation group (n =8) and UUO operation+apocynin treatment group (n =8).Either vehicle or apocynin (100 mg/kg per day) were given by gavage for 7 days after surgery.Rats were sacrificed at 7th day.ELISA was used to detect the activity of superoxide dismutase (SOD) and catalase (CAT),and the level of 8-iso-prostaglandin F2alpha (8-isoPGF2α) in renal tissue.Western blotting was used to detect the protein expressions of NADPH oxidase subunit NOX2 and NOX4,α-smooth muscle actin(α-SMA),collagen Ⅰ (COL-Ⅰ) and the level of ERK1/ 2 phosphorylation (p-ERK1/2).Results UUO rats with vehicle displayed increased oxidative stress,as measured by renal tissue 8-iso-PGF2α,accompanied with increased renal expression of NADPH oxidases (NOX2,1.5-fold and NOX4,1.7-fold,respectively),compared with sham-operated rats (P ＜0.05).Furthermore,vehicle treated UUO rats showed increased renal COL-Ⅰ and α-SMA levels,compared with sham-operated rats (P ＜ 0.05).ERK1/2 was also activated as detected by p-ERK1/2 expression in UUO rats with vehicle (P ＜ 0.05).Apocynin treatment significantly decreased renal tissue 8-iso-PGF2α level and expressions of NOX2 (-28.7％) and NOX4 (-31.0％) in UUO rats,respectively,compared with vehicle treated rats (P ＜ 0.05).And significant decrease of COL-Ⅰ (-26.4％) and α-SMA expression (-80.0％) were also observed (P ＜ 0.05).The activation of ERK1/2 in UUO rats was greatly inhibited by apocynin treatment (P ＜ 0.01).Despite the pronounced dysregulation of pro-oxidative NOXs family,no compensatory increase of antioxidative enzyme activities occurred.Conclusion The NOXs family contributes largely to the production of ROS and subsequent interstitial fibrosis after ureter ligation,and inhibition of the NOXs family may be a choice for preventing interstitial fibrosis.|
郑茜子,屈磊,唐嘉薇,等. 抑制NADPH氧化酶系统可减轻单侧输尿管梗阻大鼠肾间质纤维化[J]. 中华肾脏病杂志,2014,30(6):437-442.