地菍总黄酮对高脂饮食联合链脲佐菌素诱导糖尿病肾病小鼠脂质过氧化的影响

doi:10.16431/j.cnki.1671-7236.2025.01.033

中国畜牧兽医 ›› 2025, Vol. 52 ›› Issue (1): 364-375.doi: 10.16431/j.cnki.1671-7236.2025.01.033

• 基础兽医 • 上一篇

地菍总黄酮对高脂饮食联合链脲佐菌素诱导糖尿病肾病小鼠脂质过氧化的影响

唐雨菲^1,2,3,4, 莫烨云^1,2,3, 李笑笑^1,3, 杨秋莉^1,3, 林惠旅^1,3, 黄海芳^1,3, 林美莹^1,3, 李丽^1,2,3

1. 广西中医药大学药学院, 南宁 530200;
2. 广西中医药大学, 广西壮瑶药重点实验室, 广西壮瑶药协同创新中心, 广西壮族自治区民族药资源与应用工程研究中心, 南宁 530200;
3. 广西高校中药神经-代谢及免疫药理重点实验室, 南宁 530200;
4. 南宁市妇幼保健院药学部, 南宁 530011

收稿日期:2024-04-25 发布日期:2024-12-30
通讯作者: 李丽 E-mail:lilygxnn@163.com
作者简介:唐雨菲,E-mail:1172098099@qq.com。
基金资助:
广西中医药大学研究生教育创新计划项目(YCSW2023387);广西壮瑶药重点实验室(桂科基字[2014]32号);壮瑶药协同创新中心(桂教科研[2013]20号);广西壮族自治区民族药资源与应用工程研究中心(桂发改高技函[2020]2605号);广西中医药重点学科壮药学(GZXK-Z-20-64);广西一流学科中药学(民族药学)(桂教科研[2018]12号);2022年广西中医药大学校级科研项目(2022MS013)

Effect of Total Flavonoids from Melastoma dodecandrum Lour.on Lipid Peroxidation in Diabetic Nephropathy Mice Induced by High-fat Diet Combined with Streptozocin

TANG Yufei^1,2,3,4, MO Yeyun^1,2,3, LI Xiaoxiao^1,3, YANG Qiuli^1,3, LIN Huilyu^1,3, HUANG Haifang^1,3, LIN Meiying^1,3, LI Li^1,2,3

1. College of Pharmacy, Guangxi University of Traditional Chinese Medicine, Nanning 530200, China;
2. Research Center of Guangxi Zhuang Autonomous Region Medicine Resources and Application Engineering, Guangxi Collaborative Innovation Center of Zhuang and Yao Ethnic Medicine, Guangxi Key Laboratory of Zhuang and Yao Ethnic Medicine, Guangxi University of Traditional Chinese Medicine, Nanning 530200, China;
3. Key Laboratory of TCM Neuro-metabolism and Immunopharmacology of Guangxi Education Department, Nanning 530200, China;
4. Department of Pharmacy, Nanning Maternal & Child Health Hospital, Nanning 530011, China

Received:2024-04-25 Published:2024-12-30

摘要/Abstract

摘要： 【目的】探讨地菍总黄酮(total Flavonoids from Melastoma dodecandrum Lour.,TFMD)对糖尿病肾病(DN)小鼠脂质过氧化的作用,对TFMD在肾脏功能保护及抗脂质过氧化方面的药用价值进行探索,为瑶药地菍的临床应用研究提供试验依据。【方法】将90只SPF级雄性C57BL/6J小鼠随机分成2组:空白组(n＝10)及造模组(n＝80)。空白组小鼠给予普通饲料,造模组小鼠给予高脂饲料,连续饲养6周后,各组小鼠禁食不禁水12 h后,造模组小鼠腹腔注射链脲佐菌素(STZ)0.05 g/kg,空白组小鼠注射等剂量柠檬酸钠缓冲液,连续注射5 d后,分别在末次注射72 h及1周后测量各组小鼠空腹血糖(FBG)。将连续2次FBG≥16.7 mmol/L的小鼠随机分为5组:DN模型组(高脂饲料)、二甲双胍组(高脂饲料＋0.5 g/kg 二甲双胍)及TFMD高(高脂饲料＋1.2 g/kg TFMD)、中(高脂饲料＋0.8 g/kg TFMD)、低(高脂饲料＋0.6 g/kg TFMD)剂量组,每组10 只,连续给药10周。给药过程中观察小鼠一般生理状况,每周检测FBG。给药10周后,使用代谢笼收集各组小鼠24 h尿液,剪尾采血检测各组小鼠FBG,计算各组小鼠肾脏指数;同时检测各组小鼠尿液中尿蛋白(UP)和尿微量白蛋白(MAU)、血清尿素氮(BUN)和血清肌酐(SCr)水平及肾脏组织中谷胱甘肽(GSH)、超氧化物歧化酶(SOD)活性和丙二醛(MDA)水平。Western blotting检测各组小鼠肾脏组织Nephrin、podocin、PTGS2、ACSL4蛋白的表达量;制作各组小鼠肾脏组织HE及PAS染色切片并观察病理改变。【结果】与空白组相比,DN模型组小鼠体重、GSH和SOD水平均极显著降低(P＜0.01),FBG、肾脏指数、MAU、UP、SCr、BUN及MDA水平均极显著升高(P＜0.01),肾脏组织Nephrin、podocin蛋白表达量均极显著降低(P＜0.01),PTGS2、ACSL4蛋白表达量显著升高(P＜0.05),肾脏组织光镜下可见严重损伤,表明DN模型构建成功。与DN模型组相比,TFMD能改善DN小鼠的消瘦状况,极显著或显著降低小鼠肾脏指数及FBG(P＜0.01;P＜0.05),极显著降低小鼠体内UP、MAU、SCr、BUN水平(P＜0.01),显著提高小鼠肾脏组织Nephrin、podocin水平(P＜0.05),TFMD组小鼠肾脏组织损伤得到不同程度恢复。与DN模型组相比,TFMD显著或极显著提高小鼠肾脏组织GSH和SOD活性(P＜0.05;P＜0.01),极显著或显著降低MDA水平(P＜0.01;P＜0.05),显著或极显著下调肾脏组织中脂质过氧化相关蛋白PTGS2、ACSL4的表达(P＜0.05;P＜0.01),提升机体抗氧化能力及减缓机体脂质过氧化及其产物堆积。【结论】 TFMD可明显延缓高脂饮食联合STZ诱导小鼠DN的发生发展进程,其机制与调控脂质过氧化及氧化应激、调控PTGS2/ACSL4信号通路相关。

关键词: 地菍总黄酮; 糖尿病肾病; 脂质过氧化; 氧化应激; 脂毒性

Abstract: 【Objective】 This study was aimed to investigate the effect of total flavonoids from Melastoma dodecandrum Lour.(TFMD) on lipid peroxidation in diabetic nephropathy (DN) mice,and explore the medicinal value of TFMD in renal function protection and anti-lipid peroxidation,so as to provide experimental basis for the clinical application of Yao medicine Melastoma dodecandrum Lour. 【Method】 Ninety SPF male C57BL/6J mice were randomly divided into blank group (n＝10) and model group (n=80).Mice in blank group were given normal diet,and mice in model group were given high-fat diet for 6 weeks,after fasting for 12 h,mice in model group were intraperitoneally injected with streptozotocin (STZ) 0.05 g/kg,and mice in blank group were injected with the same dose of sodium citrate buffer.After 5 days of continuous injection,fasting blood glucose (FBG) was measured at 72 h and 1 week after the last injection.Mice with FBG≥16.7 mmol/L for two consecutive times were randomly divided into DN model group (high-fat diet),metformin group (high-fat diet＋0.5 g/kg metformin),TFMD high-dose group (high-fat diet＋1.2 g/kg TFMD),TFMD middle-dose group (high-fat diet＋0.8 g/kg TFMD) and TFMD low-dose group (high-fat diet＋0.6 g/kg TFMD),10 mice in each group.After 10 weeks of administration,the general physiological status of mice was observed during administration,and FBG was detected weekly.After 10 weeks of administration,24 h urine was collected,FBG of mice in each group was detected by cutting tail,and the kidney index were calculated. The levels of urine protein (UP) and microalbumin (MAU),serum urea nitrogen (BUN) and serum creatinine (SCr),and activities of glutathione (GSH) and superoxide dismutase (SOD) and levels of malondialdehyde (MDA) in renal tissue were detected.The expression of Nephrin,podocin,PTGS2 and ACSL4 proteins in renal tissue of mice in each group was detected by Western blotting.HE and PAS staining sections of kidney tissues of mice in each group were made and pathological changes were observed. 【Result】 Compared with blank group,the body weight,GSH and SOD activities of mice in DN model group were extremely significantly decreased (P＜0.01),FBG,kidney index,the levels of MAU,UP,SCr,BUN and MDA were extremely significantly increased (P＜0.01),the expression of Nephrin and podocin proteins in renal tissue was extremely significantly decreased (P＜0.01),the expression of PTGS2 and ACSL4 were significantly increased (P＜0.05),and severe damage was observed in renal tissue under light microscope,indicating that the DN model was constructed successfully.TFMD could improve the weight loss of DN mice,significantly reduce the kidney index and FBG (P＜0.05 or P＜0.01),reduce the levels of UP,MAU,SCr and BUN in DN mice (P＜0.01),and increase the levels of Nephrin and podocin in renal tissue (P＜0.05).The damage of kidney tissue in TFMD group was recovered to different degrees.TFMD also increased the levels of GSH and SOD in DN kidney tissue (P＜0.05 or P＜0.01),decreased the level of MDA (P＜0.05 or P＜0.01),down-regulated the expression of lipid peroxidation-related proteins PTGS2 and ACSL4 in kidney tissue (P＜0.05 or P＜0.01),improved the body’s antioxidant capacity and slowed down the body’s lipid peroxidation and its product accumulation. 【Conclusion】 TFMD could significantly delay the occurrence and development of DN induced by high-fat diet combined with STZ,and its mechanism was related to the regulation of lipid peroxidation and oxidative stress,and the regulation of PTGS2/ACSL4 signaling pathway.

Key words: total flavonoids from Melastoma dodecandrum Lour.; diabetic nephropathy; lipid peroxidation; oxidative stress; lipotoxicity

中图分类号:

S853.74

唐雨菲, 莫烨云, 李笑笑, 杨秋莉, 林惠旅, 黄海芳, 林美莹, 李丽. 地菍总黄酮对高脂饮食联合链脲佐菌素诱导糖尿病肾病小鼠脂质过氧化的影响[J]. 中国畜牧兽医, 2025, 52(1): 364-375.

TANG Yufei, MO Yeyun, LI Xiaoxiao, YANG Qiuli, LIN Huilyu, HUANG Haifang, LIN Meiying, LI Li. Effect of Total Flavonoids from Melastoma dodecandrum Lour.on Lipid Peroxidation in Diabetic Nephropathy Mice Induced by High-fat Diet Combined with Streptozocin[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 364-375.

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地菍总黄酮对高脂饮食联合链脲佐菌素诱导糖尿病肾病小鼠脂质过氧化的影响

Effect of Total Flavonoids from Melastoma dodecandrum Lour.on Lipid Peroxidation in Diabetic Nephropathy Mice Induced by High-fat Diet Combined with Streptozocin

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