文献类型：期刊文献

中文题名：基于PI3K/Akt/mTOR信号通路探讨萆薢分清颗粒对抗单侧输尿管梗阻大鼠肾纤维化影响的研究

英文题名：Study on the effect of Bixie Fenqing Granules on renal fibrosis in UUO rats via the PI3K/Akt/mTOR signaling pathway

作者：魏锦慧[1];马鸿斌[1];翟书玲[2];李根生[2];何彩苹[1];马雪莉[1];张莲荷[1]

第一作者：魏锦慧

机构：[1]甘肃中医药大学附属医院,甘肃兰州730020;[2]甘肃中医药大学,甘肃兰州730000

第一机构：甘肃中医药大学第二附属医院

年份：2026

卷号：27

期号：2

起止页码：144

中文期刊名：山西中医药大学学报

外文期刊名：Journal of Shanxi University of Chinese Medicine

基金：甘肃省自然科学基金项目(21JR7RA577);甘肃省名中医传承马鸿斌工作室建设项目(甘财社[2022]105号);甘肃中医药大学附属医院创新基金一般项目(gzfy-2023-03)。

语种：中文

中文关键词：UUO模型大鼠;萆薢分清颗粒;PI3K/Akt/mTOR信号通路

外文关键词：UUO model rats;Bixie Fenqing Granules(Rhizoma Dioscoreae Granules for Clearing Turbid Urine);PI3K/Akt/mTOR signaling pathway

摘要：目的:建立单侧输尿管梗阻(UUO)大鼠模型,观察萆薢分清颗粒对UUO大鼠肾纤维化(RF)相关蛋白表达的影响,探讨其抗RF的作用机制。方法:选用SPF级雄性8周龄SD大鼠60只,随机分为空白组、模型组、氯沙坦钾组、萆薢分清颗粒高、中、低剂量组,每组10只。药物干预过程中观察大鼠一般状况,第4周采集各组大鼠血清、尿液,检测24 h-尿蛋白定量(24 h-UPQ)、血肌酐(Scr)、血尿素氮(BUN)水平,肌氨酸氧化酶法检测尿肌酐含量,ELISA法检测尿微量白蛋白含量,计算尿微量白蛋白/肌酐(ACR);肾组织标本苏木精-伊红(HE)染色、马松(Masson)染色,观察各组大鼠肾组织纤维化程度;蛋白免疫印迹法(Western Blot)、免疫组化法检测肾组织中磷脂酰肌醇3-激酶(PI3K)、丝氨酸-苏氨酸蛋白激酶B(Akt)、哺乳动物雷帕霉素靶蛋白(mTOR)的表达水平。结果:(1)与空白组比较,模型组大鼠精神倦怠,一般状况差;24 h-UPQ、ACR、Scr、BUN水平均明显升高,差异均有统计学意义(P<0.05,P<0.01);HE染色示肾小管萎缩、结构破坏伴有空泡变性,间质纤维细胞增生,并可见炎性细胞浸润,肾小球结构破坏固缩;Masson染色可见大量蓝色胶原纤维沉积,RF程度高;免疫组化、Western Blot检测均提示模型组大鼠肾组织PI3K、Akt、mTOR表达明显增强,差异均有统计学意义(P<0.05,P<0.01)。(2)给药4周后,与模型组比较,各干预组大鼠一般状况均有不同程度好转,其中氯沙坦钾组、萆薢分清颗粒高剂量组改善程度最好;HE染色示氯沙坦钾组、萆薢分清颗粒高剂量组病理损伤较模型组明显好转;Masson染色示除萆薢分清颗粒低剂量组外,其余干预组蓝色胶原纤维面积均有不同程度减少,纤维化程度有所改善。与模型组比较,萆薢分清颗粒高、中剂量组、氯沙坦钾组大鼠24 h-UPQ、ACR、Scr、BUN水平明显降低(P<0.05,P<0.01),萆薢分清颗粒低剂量组大鼠Scr、BUN水平降低,差异均有统计学意义(P<0.05)。免疫组化、Western Blot检测示与模型组比较,氯沙坦钾组、萆薢分清颗粒高、中剂量组大鼠肾组织PI3K、Akt、mTOR表达明显降低,差异均有统计学意义(P<0.05,P<0.01),低剂量组大鼠肾组织PI3K、Akt表达明显降低(P<0.05,P<0.01),且萆薢分清颗粒高剂量组大鼠肾组织PI3K表达水平明显低于中、低剂量组,差异均有统计学意义(P<0.05)。(3)萆薢分清颗粒高剂量组与氯沙坦钾组比较,两组在改善大鼠一般状况、降低24 h-UPQ、ACR、Scr、BUN水平,以及减轻肾组织病理损伤和纤维化方面效果相当,差异均无统计学意义(P>0.05)。免疫组化、Western Blot检测均提示两组抑制UUO大鼠肾组织PI3K、Akt蛋白表达效果相当,差异均无统计学意义(P>0.05),高剂量组mTOR蛋白表达强度低于氯沙坦钾组,差异均有统计学意义(P<0.05)。结论:萆薢分清颗粒通过降低UUO大鼠24 h-UPQ、ACR、Scr、BUN水平起到减少蛋白尿、减轻肾组织病理损伤、改善肾功能、保护肾脏的作用;通过下调肾组织中RF相关蛋白PI3K、Akt、mTOR的表达,延缓RF进展。
Objective:To establish a unilateral ureteral obstruction(UUO)rat model and investigate the effect of Bixie Fenqing Granules(Rhizoma Dioscoreae Granules for Clearing Turbid Urine)on the expression of renal fibrosis(RF)-related proteins in UUO rats,thereby exploring its mechanism against RF.Methods:Sixty SPF-grade male SD rats aged 8 weeks were randomly divided into six groups(n=10 per group):blank group,model group,losartan potassium group,and high-,medium-,and low-dose Bixie Fenqing Granules groups.The general condition of the rats was observed during drug intervention.At week 4,serum and urine samples were collected to measure 24-hour urine protein quantification(24 h-UPQ),serum creatinine(Scr),and blood urea nitrogen(BUN)levels.Urinary creatinine content was detected by the sarcosine oxidase method,and urinary microalbumin was measured by ELISA to calculate the urinary microalbumin-tocreatinine ratio(ACR).Renal tissue specimens were stained with hematoxylin-eosin(HE)and Masson staining to observe the degree of renal fibrosis.The expression levels of PI3K,Akt,and mTOR proteins in renal tissues were detected by Western blot and immunohistochemistry.Results:(1)Compared with the blank group,the model group showed lethargy and poor general condition;24 h-UPQ,ACR,Scr,and BUN levels were significantly increased(P<0.05,P<0.01).HE staining revealed renal tubular atrophy,structural damage with vacuolar degeneration,interstitial fibroblast proliferation,inflammatory cell infiltration,and glomerular structural destruction and consolidation.Masson staining showed extensive blue collagen fiber deposition,indicating severe RF.Immunohistochemistry and Western blot results indicated significantly enhanced expression of PI3K,Akt,and mTOR in the model group(P<0.05,P<0.01).(2)After 4 weeks of intervention,all treatment groups showed improved general conditions,with the losartan potassium and high-dose Bixie Fenqing Granules groups exhibiting the most significant improvement.HE staining indicated marked alleviation of pathological damage in the losartan potassium and high-dose Bixie Fenqing Granules groups compared to the model group.Masson staining showed reduced blue collagen fiber area and improved fibrosis in all intervention groups except the low-dose Bixie Fenqing Granules group.Compared with the model group,the high-and medium-dose Bixie Fenqing Granules groups and the losartan potassium group had significantly lower 24 h-UPQ,ACR,Scr,and BUN levels(P<0.05,P<0.01),while the low-dose group showed reduced Scr and BUN levels(P<0.05).Immunohistochemistry and Western blot results demonstrated that the losartan potassium group and the high-and medium-dose Bixie Fenqing Granules groups had significantly lower expression of PI3K,Akt,and mTOR in renal tissues(P<0.05,P<0.01).The low-dose group showed reduced PI3K and Akt expression(P<0.05,P<0.01),and the high-dose group had significantly lower PI3K expression than the medium-and low-dose groups(P<0.05).(3)Compared with the losartan potassium group,the high-dose Bixie Fenqing Granules group showed equivalent effects in improving general condition,reducing 24 h-UPQ,ACR,Scr,and BUN levels,and alleviating renal pathological damage and fibrosis(P>0.05).Immunohistochemistry and Western blot results indicated that the two groups equally suppressed PI3K and Akt protein expression in UUO rat renal tissues(P>0.05),while the high-dose group had lower mTOR expression than the losartan potassium group(P<0.05).Conclusion:Bixie Fenqing Granules reduce proteinuria,alleviate renal pathological damage,improve renal function,and protect the kidneys by lowering 24 h-UPQ,ACR,Scr,and BUN levels in UUO rats.They delay RF progression by downregulating the expression of RF-related proteins PI3K,Akt,and mTOR in renal tissues.