抗纤灵方对肾纤维化小鼠肾组织CD68、CD45、VCAM-1表达的影响

Effect of Kangxianling Fang(抗纤灵方) on Expression of CD68,CD45,and VCAM-1 in Renal Tissue of Renal Fibrosis Mice

目的探讨抗纤灵方治疗肾纤维化的可能作用机制。方法将测得血压<120 mm Hg的小鼠随机分为正常组、手术组、模型组、抗纤灵组、蒙诺组,每组12只。除正常组、手术组外,其余各组小鼠采用单侧肾切除及微渗透泵灌注血管紧张素Ⅱ(AngⅡ)建立肾纤维化小鼠模型,手术组仅行单侧肾切除手术后微渗透泵灌注0.9%氯化钠溶液。在植入微渗透泵后,抗纤灵组给予抗纤灵方2.3 g/100 g灌胃,蒙诺组给予福辛普利钠片0.03 g/100 g灌胃,正常组、手术组和模型组给予自来水灌胃。各组均每日1次,每次0.5 ml,灌胃4周。4周后采用免疫组织化学法和Western Blot法检测小鼠肾组织CD68、CD45、血管内皮细胞黏附分子1(VCAM-1)表达。结果免疫组织化学法检测结果显示,与模型组比较,抗纤灵组和蒙诺组小鼠肾组织CD68、CD45、VCAM-1表达灰度值均有不同程度下降(P<0.05或P<0.01);抗纤灵组小鼠肾组织VCAM-1表达灰度值高于蒙诺组(P<0.05)。Western Blot法检测结果显示,与模型组比较,抗纤灵组和蒙诺组小鼠肾组织CD68表达降低(P<0.01),CD45表达与模型组比较差异无统计学意义(P>0.05);蒙诺组小鼠肾组织VCAM-1表达低于模型组和抗纤灵组(P<0.05或P<0.01);而抗纤灵组VCAM-1表达与模型组比较差异无统计意义(P>0.05)。结论抗纤灵方可降低肾纤维化小鼠肾组织CD68、CD45、VCAM-1的表达,这可能是其治疗肾纤维化的作用机制之一。

Objective To explore the possible mechanism of Kangxianling Fang（抗纤灵方） in the treatment of renal fibrosis. Methods Mice with blood pressure 120 mm Hg were randomly divided into normal group,surgical group,model group,Kangxianling Fang group,and fosinopril group,with 12 mice in each group. Except for the normal group and surgical group,mice in the other groups were treated with unilateral nephrectomy and micro-osmotic pump perfusion of angiotensin Ⅱ（Ang Ⅱ） to establish model of renal fibrosis. The surgical group was only given unilateral nephrectomy,and micro-osmotic pump perfusion of 0. 9% sodium chloride solution. After implantation of the micro-osmotic pump,Kangxianling Fang group was given intragastric administration of 2. 3 g/100 g Kangxianling Fang. The fosinopril group was given 0. 03 g/100 g fosinopril tablets by gavage. The normal group,surgical group and model group were given tap water gavage. Each group was given once a day,0. 5 ml each time for 4 weeks. Afterwards,the expressions of CD68,CD45,and vascular cell adhesion molecule-1（VCAM-1） in the kidney of mice were detected by immunohistochemistry and Western Blot. Results Immunohistochemistry results showed that compared with the model group,the expression gray value of CD68,CD45,and VCAM-1 in the renal tissues of the mice in Kangxianling Fang group and fosinopril group were all decreased to varying degrees（P〈0. 05 or P〈0. 01）. The expression gray value of VCAM-1 in renal tissue of mice in Kangxianling Fang group was higher than fosinopril group（P〈0. 05）. Western Blot assay results showed that compared with the model group,the expression of CD68 in the renal tissue of mice in Kangxianling Fang group and fosinopril group was decreased（P〈0. 01）,and there was no significant difference in the expression of CD45 compared with model group（P〈0. 05）. The expression of VCAM-1 in the renal tissue of mice in fosinopril group was lower than that in the model group and Kangxianling Fang group（P〈0. 05 or P〈0. 01）,but the difference was not statistically significant in the expression of VCAM-1 of Kangxianling Fang group when compared with model group（P〈0. 05）. Conclusion Kangxianling Fang could reduce the expression of CD68,CD45 and VCAM-1 in the renal tissue of renal fibrosis mice,which may be one of the mechanisms in treating renal fibrosis.