加味八正散对慢性肾小球肾炎大鼠炎症因子及肾脏组织病理改变影响的实验研究

Experimental Research on the Effect of JiaWeiBaZhengSan on Inflammatory Factors in Chronic Glomerulonephritis Rats and Histopathological Changes of Renal Tissues

目的研究加味八正散对慢性肾小球肾炎(CGN)大鼠炎症因子及肾脏组织病理改变的影响。方法按照随机原则将90只大鼠分为6组:空白组,模型组,肾炎康复片组,加味八正散高、中、低剂量组。于实验开始前一日及处死大鼠前一日收集24 h尿液,检测尿蛋白定量。实验结束时,处死所有大鼠,用酶联免疫吸附测定(ELISA)法检测肿瘤坏死因子α(TNF-α)、γ干扰素(IFN-γ)、白介素4(IL-4)、白介素6(IL-6)、白介素10(IL-10)的表达。处死大鼠时摘取肾脏行HE染色,光镜下观察大鼠肾脏病理改变并分级。结果造模前,各组24 h尿蛋白定量差别不大;治疗后,模型组24 h尿蛋白定量远高于空白组(P<0.05)。与模型组相比,肾炎康复片组和加味八正散高、中、低剂量组24 h尿蛋白定量均降低(P<0.05);与空白组相比,模型组血清中IFN-γ、TNF-α含量明显增加,IL-4、IL-6、IL-10含量明显下降(P<0.05);与模型组相比,肾炎康复片组和加味八正散高、中、低剂量组血清中IFN-γ、TNF-α含量都有所下降,而IL-4、IL-6、IL-10含量均明显上升(P<0.05)。空白组肾脏组织病理改变为0级,肾小球、肾小管形态正常,结构清晰,没有病理表现;模型组肾脏组织病理改变为Ⅲ级,可见明显的肾小管结构紊乱、肿胀、变性,大量炎性细胞浸润,肾小管上皮细胞脱落且基底膜增厚,上皮细胞变性坏死,出现明显的间质纤维化;肾炎康复片组肾脏组织病理改变为Ⅰ级,与模型组相比,肾小管炎性细胞浸润明显减轻,萎缩及纤维化程度改善;加味八正散低剂量组肾脏组织病理改变为Ⅱ级,与模型组相比,肾小管炎性细胞浸润程度稍有减轻,萎缩及纤维化程度得到改善,但有系膜区轻度增宽;加味八正散中剂量组肾脏组织病理改变为Ⅰ级,与模型组相比,肾小管炎性细胞浸润程度稍有减轻,萎缩及纤维化程度得到改善;加味八正散高剂量组肾脏组织病理改变为0级,与模型组相比,肾小球、肾小管形态结构基本恢复正常,肾小管无炎性细胞浸润,萎缩及纤维化程度明显改善。结论经加味八正散治疗后,肾小球肾炎大鼠24 h尿蛋白定量降低,说明加味八正散具有治疗效果,且高剂量组治疗效果最好;大鼠血清中TNF-α、IFN-γ含量下降,而IL-4、IL-6、IL-10含量升高,说明加味八正散对于慢性肾小球肾炎疗效较好,能够提高炎症介质水平,具有抗炎作用。同时减轻大鼠肾脏病理损伤,对肾脏具有保护作用。

Objective To research the effect of Jia Wei Ba Zheng San on inflammatory factors in Chronic Glomerulonephritis(CGN)rats and histopathological changes of renal tissues.Methods According to random principle 90 rats were divided into six groups:blank group,model group,Shen Yan Kang Fu Pian group,and the high,middle and low dose groups of Jia Wei Ba Zheng San.24-hour urine was collected before the day of the experiment started and the day before the rats’execution to measure the urine protein quantification.At the end of the experiment,all the rats were executed and the expressions of tumor necrosis factorα(INE-α),γinterferon(IFN-γ),interleukin 4(IL-4),interleukin 6(IL-6)and interleukin 10(IL-10)were measured by enzyme-linked immunosorbent assay(ELISA).The HE renal tissues of the rats were removed in execution for staining,and the histopathological changes of renal tissues were observed and graded under light microscopy.Re suits Before modeling,the 24 h urine protein quantification of each group was not significantly different;after treatment,the 24 h urine protein quantification of the model group was much higher than that of the blank group(P<0.05).Compared with the model group,the 24 h urine protein quantification was decreased both in ShenYan Kang Fu Pian group and the high,medium and low dose groups of Jia Wei Ba Zheng San(P<0.05);compared with the blank group,the serum levels of IFN-γand TNF-αwere significantly increased and the levels of IL-4,IL-6 and IL-10 were significantly decreased in the model group(P<0.05);compared with the model group,the serum levels of IFN-γand TNF-αwere decreased in ShenYan Kang Fu Pian group and the high,medium and low dose groups of Jia Wei Ba Zheng San(P<0.05).Compared with the model group,the serum levels of IFN-γand TNF-αdecreased,while the levels of IL-4,IL-6 and IL-10 increased significantly(P<0.05).The histopathological changes of renal tissues in the blank group were grade 0,with normal glomerular and tubules,clear structure,and no pathological manifestations;the histopathological changes in renal tissues of the model group were grade III,the tubules had obvious structural disorder,swelling,degeneration,massive inflammatory cell infiltration,the renal tubular epithelial cells detached and the basement membrane thickened,the epithelial cells were degenerative and necrotic,and existed obvious interstitial fibrosis;the histopathological changes of renal tissues in ShenYan Kang Fu Pian group were grade I.Compared with the model group,the inflammatory cell infiltration of tubules was significantly reduced,and the degrees of atrophy and fibrosis were improved;the histopathological changes in renal tissues in the low-dose group of Jia Wei Ba Zheng San were grade II.Compared with the model group,the degree of inflammatory cell infiltration in tubules was slightly reduced,and the degrees of atrophy and fibrosis were improved,but there was a mild widening of the mesangial cell;the histopathological changes in renal tissues in the middle-dose group of Jia Wei Ba Zheng San were grade I,compared with the model group,the degree of inflammatory cell infiltration in tubules was slightly reduced,and the degrees of atrophy and fibrosis were improved;the histopathological changes in renal tissues in the high-dose group of Jia Wei Ba Zheng San were grade 0,compared with the model group,the morphological structures of glomerular and tubules were basically normalized,and there was no inflammatory cell infiltration in tubules,and the degrees of atrophy and fibrosis were significantly improved.Conclusio n The 24 h urine protein quantification of rats with glomerulonephritis decreased after treatment with Jia Wei Ba Zheng San indicated Jia Wei Ba Zheng San had effect on treatment,and the high-dose group had the best effect.The levels of TNF-αand IFN-γin the serum of rats decreased while the levels of IL-4,IL-6 and IL-10 increased,indicated the efficacy of Jia Wei Ba Zheng San is better for chronic glomerulonephritis(CGN),which can increase the inflammatory mediators levels and had anti-inflammatory effects,it reduced the pathological damage in renal tissues of rats and protected the renal tissues at the same time.