益气养阴活血方对IgA肾病小鼠结肠病理及免疫的影响

Effect of Yiqi Yangyin Huoxue Recipe on colon pathology and immunity in mice with IgA nephropathy

目的观察益气养阴活血方对IgA肾病小鼠结肠病理与血清Gd-IgA1/IL17/BAFF表达水平相关性的影响,探讨其疗效机制。方法采用牛血清白蛋白(BSA)灌胃+皮下注射四氯化碳(CCl_(4))+尾静脉注射脂多糖(LPS)的方法建立IgA肾病小鼠模型。BLBAC小鼠60只,适应性喂养1周后随机分为5组:空白组、模型组、泼尼松片组(激素组)、益气养阴活血方中药等剂量组(中药组)、益气养阴中药高剂量组(中高组),生化检测24h尿蛋白定量(24hUTP)、谷丙转氨酶(ALT)、血清白蛋白(ALB)、尿素氮(BUN)、血肌酐(Scr)水平;HE染色观察各组肾组织及结肠病理改变,免疫荧光观察肾小球系膜区IgA沉积;ELISA法检测血清异常糖基化IgA1(Gd-IgA1)、白介素17(IL-17)及B细胞活化因子(BAFF)水平。结果(1)与模型组比较,激素组及中药两组ALB水平显著升高,24hUTP、ALT、BUN、Scr水平降低(P﹤0.05),血清Gd-IgA1、IL-17、BAFF水平均明显降低(P﹤0.05);与激素组比较,中药两组无统计学差异(P>0.05);(2)模型组肾脏病理观察,光镜下可见肾小球局灶系膜细胞和系膜基质有不同程度的增生,灶性轻度缺血皱缩,肾小管、间质片状和灶性慢性炎症,免疫荧光见强阳性的IgA沉积。与模型组相比,激素组及中药两组光镜下肾小球、肾小管病变及免疫荧光IgA沉积减轻;(3)模型组结肠病理观察可见肠绒毛缺损,中央乳糜管明显扩张,不同程度的上皮细胞脱落,肠绒毛较细短,淋巴细胞增多,伴有大量炎性细胞浸润。激素组及中药两组结肠病理较模型组减轻;(4)上述指标及病理观察,激素组和中药两组间、中药组及中高组之间均无明显差异。结论益气养阴活血方可能通过抑制IgA肾病小鼠肠黏膜免疫炎症反应,减少血清BAFF、IL-17及Gd-IgA1的产生,改善肾脏病理变化,减少蛋白尿,保护肾功能;中药组与激素组疗效相当。

Objective To observe the effect of Yiqi Yangyin Huoxue Recipe on the correlation between colon pathology and serum Gd-IgA1/IL17/BAFF expression levels in mice with IgA nephropathy and to explore its efficacy mechanism.Methods A mouse model of IgA nephropathy was established by gavage of bovine serum albumin(BSA)+subcutaneous injection of carbon tetrachloride(CCL_(4))+tail vein injection of lipopolysaccharide(LPS),randomly divided into blank group,model group,Prednisone group(hormone group),equal-dose group of Chinese medicine(Chinese medicine group)and high-dose group of Chinese medicine(medium-high group),biochemical tests for 24 h urine protein(24 hUTP),alanine aminotransferase(ALT),serum albumin(ALB),urea nitrogen(BUN),serum creatinine(Scr)level;Pathological changes of renal tissues and colon were observed by HE staining.Immunofluorescence was used to observe IgA deposition in glomerular mesangial area.ELISA was used to detect the serum levels of abnormal glycosylated IgA1(Gd-IgA1),interleukin 17(IL-17)and B cell activating factor(BAFF).Results(1)Compared with the model group,the levels of ALB in the hormone group and the two groups of traditional Chinese medicine increased significantly.24 hUTP,ALT,BUN,Scr levels were decreased(P<0.05),serum Gd-IgA1/,IL-17,BAFF levels were significantly decreased(P<0.05).Compared with hormone group:there is no statistical difference between the two groups of traditional Chinese medicine.(P>0.05);(2)In the model group,renal pathology showed that the glomerular focal mesangial cells and mesangial matrix had different degrees of proliferation,focal mild ischemic shrinkage,renal tubules,interstitial flaky and focal chronic inflammation,immunofluorescence showed the strong positive IgA deposition.Compared with the model group,the glomerulus and renal tubule lesions and immunofluorescence IgA deposition were reduced under the light microscope in the hormone group and the Chinese medicine groups;(3)In the model group,the colon pathology showed intestinal villi defect,the central lacteal was obviously dilated,the epithelial cells were shed to varying degrees,the intestinal villi were shorter,the lymphocytes increased,accompanied by a large number of inflammatory cell infiltrations.The pathological changes of colon in the hormone group and Chinese medicine groups were less than that in the model group;(4)The above indicators and pathology showed no significant difference between hormone group and two traditional Chinese medicine groups,between Chinese medicine group and medium-high group.Conclusion Yiqi Yangyin Huoxue Recipe may inhibit the intestinal mucosal immune inflammatory response in IgA nephropathy mice,reduce the production of serum BAFF,IL-17 and Gd-IgA1,improve renal pathology,reduce proteinuria,and protect renal function;The effect of traditional Chinese medicine group and hormone group is equal.