| 郎茜.杜仲多糖对糖尿病大鼠肾脏损伤的保护作用及机制研究[J].安徽医药,待发表. |
| 杜仲多糖对糖尿病大鼠肾脏损伤的保护作用及机制研究 |
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| 投稿时间：2023-12-23 录用日期：2024-04-16 |
| DOI： |
| 中文关键词: 杜仲多糖 糖尿病 肾脏损伤 氧化应激 凋亡 |
| 英文关键词: |
| 基金项目:云南省教育厅科学研究基金项目（2021J1114） |
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| 摘要点击次数: 85 |
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| 中文摘要: |
| 摘要：
目的 明确杜仲多糖对糖尿病大鼠肾脏功能损伤的保护作用并初步探究其作用机制。方法 选取6-8周龄雄性SD大鼠60只，按照65 mg/kg的剂量腹腔注射链脲佐菌素(STZ)。72小时后，用尿糖试纸检测尿糖强度，尾静脉采血，测定空腹血糖(FBG)，尿糖呈强阳性且FBG≥16.7mmol/L，即认为造模成功。将成功建模的SD大鼠，随机分成模型组和杜仲多糖低剂量实验组、中剂量实验组、高剂量实验组及二甲双胍组，每组12只；另选12只健康SD大鼠作为正常组。正常组与模型组每日单次灌胃给予0.9%生理盐水；低、中、高剂量实验组分别按50 mg/kg、100 mg/kg、200mg/kg剂量，每日单次灌胃给予杜仲多糖；二甲双胍组按200mg/kg剂量，每日单次灌胃给予二甲双胍。连续给药干预7天后，每组随机选取6只大鼠进行研究，剩下大鼠继续给药干预7天，取材并对大鼠FBG、体重、肾脏指数等指标进行分析。检测杜仲多糖对各组大鼠FBG、体重、肾脏指数的影响；用酶联免疫法(ELISA)对血清尿素氮(BUN)、肾匀浆液中丙二醛(MDA)含量、总超氧化物歧化酶(T-SOD)以及谷胱甘肽过氧化物酶(GSH-Px)活力等指标予以检测。通过苏木精-伊红染色(HE)观察大鼠肾组织受损的病理情况。采用蛋白免疫印迹法(WB)对各组大鼠肾组织转化生长因子-β1(TGF-β1)、p38丝裂原活化蛋白激酶( p38 MAPK)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)等因子的表达情况进行检测。结果 与模型组相比，低、中、高剂量杜仲多糖实验组大鼠体重显著增加，高剂量实验组及二甲双胍组的大鼠肾脏指数显著降低。同时，中、高剂量杜仲多糖实验组大鼠FBG、BUN显著降低。病理学结果显示，杜仲多糖的干预可以缓解糖尿病大鼠肾脏病变程度且呈剂量依赖，剂量越高对大鼠肾小管上皮细胞肿胀空泡化的缓解作用越强。ELISA检测结果表明，杜仲多糖的干预可以提高肾组织抗氧化酶T-SOD和过氧化物分解酶GSH-Px活力，并降低MDA含量。WB实验结果表明，杜仲多糖能够下调肾组织TGF-β1、p38 MAPK以及Caspase-3等因子的表达。结论 杜仲多糖可以显著降低糖尿病大鼠的血糖，其改善糖尿病大鼠体重、肾脏指数、血清BUN含量、抗氧化能力与二甲双胍效果相当，且呈剂量、时间依赖。同时，杜仲多糖可减轻肾小球内淤血和肾小管上皮细胞空泡化，从而减轻糖尿病所导致的肾脏微血管损伤，对糖尿病肾病的早期发展具有一定的正向干预作用。随着杜仲多糖剂量的增加，TGF-β1、p38MAPK、细胞凋亡重要效应因子Caspase-3的表达水平下调越明显。故认为杜仲多糖能缓解糖尿病对肾脏的损伤，发挥对糖尿病肾病的治疗作用，作用机制可能与抗氧化应激及干预细胞凋亡有关。 |
| 英文摘要: |
| Abstract： Objectives To clarify the protective effect and potetial mechanism of Eucommia ulmoides polysaccharide on renal damage of diabetic rats. Methods Diabetic SD rats were randomly divided into model group, low-, medium- and high-dose of Eucommia ulmoides polysaccharide experimental groups, the metformin group was severed as a positive control. Each group had 12 rats. Another 12 healthy SD rats were used as normal group. The normal and model group rats were given 0.9% normal saline by intragastric administration once a day. The low-, medium- and high-dose experimental groups were intragastrically administrated with Eucommia ulmoides polysaccharide once a day at the dose of 50 mg/kg, 100 mg/kg and 200 mg/kg, respectively. Metformin group was given metformin by gavage at a dose of 200 mg/kg daily. After 7 days of continuous administration, 6 rats were randomly selected from each group for analysis, while the remaining rats were continued to be administered for another 7 days. Serum blood urea nitrogen (BUN), content of malondialdehyde (MDA) in kidney homogenate, total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) activity were measured by enzyme-linked immunosorbent assay (ELISA). Hematoxylin-eosin staining (HE) was used to observe the pathological conditions of rat kidney tissue damage. Western blotting (WB) was used to detect the expression level of transforming growth factor-β1 (TGF-β1), p38 mitogen activated protein kinase (p38-MAPK) and cysteine aspartate specific protease-3 (Caspase-3) in rat kidney of each group rats. Results Compared with the model group, the weight of rats in low, medium and high-dose groups of Eucommia ulmoides polysaccharide increased significantly, and the renal index in the high-dose experimental group and metformin group decreased significantly. Meanwhile, compared with model group, FBG and BUN of rats in the medium and high-dose experimental group were significantly decreased. The pathological results showed that the intervention of Eucommia ulmoides polysaccharide could alleviate the degree of renal lesions in diabetic rats in a dose-dependent manner, relieving the swelling and vacuolization of rat renal tubular epithelial cells. ELISA test showed that the intervention of Eucommia ulmoides polysaccharide could increase the activities of antioxidant enzymes T-SOD and GSH-Px in renal tissue, and reduce the content of MDA. WB results showed that Eucommia ulmoides polysaccharide could down-regulate the protein level of TGF-β1, p38 MAPK and Caspase-3 in renal tissue. Conclusions Eucommia ulmoides polysaccharide could significantly reduce FBG, kidney index, serum BUN content, while improve the body weight and antioxidant capacity of diabetic rats. The effect of Eucommia ulmoides polysaccharide is comparable to that of Metformin in a dose-and time-dependent manner. Furthermore, we found that Eucommia ulmoides polysaccharide could reduce the internal glomerular stasis and renal tubular epithelial cell vacuolation, thus, reduced the renal microvascular injury caused by diabetes mellitus. Eucommia ulmoides polysaccharide showed a positive intervention effect on the early development of DN, it surpressed protein level of TGF- β1, p38 MAPK and Caspase-3. Therefore, Eucommia ulmoides polysaccharide exhibits protective effect on renal damage of diabetic rats and play a role in the treatment of diabetic nephropathy. The mechanism is probably related to anti-oxidative stress and interference of cell apoptosis. |
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