Effects of kangxianlin on Extra cellular Matrix Depodition in 5/6 nephrectomized inducedchronic renal failure mice kidney tissue
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摘要:
目的探讨抗纤灵对5/6肾切除诱导的慢性肾脏纤维化小鼠模型肾组织细胞外基质(ECM)的影响。 方法C57BL/6J雄性小鼠，采用5/6肾切除制备慢性肾脏纤维化模型，2周后根据肌酐水平把手术组小鼠分为模型组，雷帕霉素和抗纤灵组，同时设立假手术组，每组10只，雷帕霉素，抗纤灵组分别给予雷帕霉素(0.00 016 g/kg)，抗纤灵(0.4 g/(kg·d)灌胃治疗，模型组和假手术组等量蒸馏水，连续用药4周后处死小鼠，免疫荧光法测定肾组织I型胶原(ColⅠ)，Ⅲ型胶原(Col Ⅲ)，α-平滑肌肌动蛋白(α-SMA)纤维化面积；RT-PCR法检测肾组中，Co Ⅰ，Col Ⅲ，α-SMAmRNA的表达。 结果与假手组比较，模型组Co Ⅰ，Col Ⅲ，α-SMA胶原纤维化面积比值明显增加，Co Ⅰ，Col Ⅲ，α-SMAmRNA显著增高(P < 0.01)，与模型组比较，雷帕霉素，抗纤灵组胶原纤维面积比值，mRNA表达水平均有所降低(P < 0.01)。雷帕霉素与抗纤灵组比较，两者之间无显著性差异。 结论抗纤灵具有雷帕霉素类似的作用，能够通过抑制小鼠肾组织Co Ⅰ，Col Ⅲ，α-SMAmRNA的表达，减少Co Ⅰ，Col Ⅲ，α-SMA纤维化面积，对肾纤维化有一定的改善作用。 Abstract:ObjectiveTo determine effects of Kangxianling decoction (KXLD) on Extracellular Matrix (ECM) Depodition in 5/6 nephrectomized-induced chronic renal failure mice kidney tissue. MethodsSPF C57BL/6J male mice weighing18-22g, divided into sham group and surgrey group, establish an model of chronic renal failure caused by 5/6 nephrectomized. Two weeks later after surgrey, according to serum creatinine(Scr) level, the mices were divided into model group, KXLD and rapamycin (RAP) group, so Scr level between the surgrey groups was no significant difference(P>0.05). KXLD and RAP were gavaged respectively according to 0.4 g/kg 0.00016 g/kg/day. sham and model group are the same amount of distilled water. After 4 weeks of treatment, Collagen area and mRNA expression of Collagen Type I(Co Ⅰ), Collagen Type Ⅲ (Col Ⅲ), α-smooth muscle actin, (α-SMA)were Observed by immunofluorescence and real-time PCR method. ResultsCompared with sham group, the levels Collagen area and mRNA expression of Co Ⅰ, Co Ⅲ, α-SMA increased significantly in the model group(P < 0.01), Compared with model group, the levels Collagen area and mRNA expression of Co Ⅰ, Col Ⅲ, α-SMA decreased significantly in the KXLD and RAP group (P < 0.01).Compared with KXLD and RAP, there have no different (P>0.05). ConclusionKXLD is able to decreased the level of Collagen area, inhibited mRNAexpression of Co Ⅰ, Col Ⅲ, α-SMAand have the similar effection as RAP. -
Key words:
- Kangxianling decoction /
- chronic renal fibrosis /
- ECM /
- Co Ⅰ /
- Col Ⅲ /
- α-SMA
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表 1 各组小鼠肾组织ColⅡ，Col Ⅲ，α-SMA相对荧光面积比较(n=6)
组别 Co Ⅰ Col Ⅲ α-SMA 假手术组 3.46±0.36 4.04±0.24 3.17±0.59 模型组 20.89±1.97** 22.06±1.80** 14.30±0.80** 雷帕霉素组 12.25±1.20## 12.74±1.27## 10.10±0.97## 抗纤灵组 12.01±0.97## 13.75±1.30## 11.33±1.56## *P < 0.05 vs假手术组; **P < 0.05 vs假手术组; #P < 0.05vs模型组; ##P < 0.01 vs模型组. 
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表 2 各组肾组织ColⅡ，Col Ⅲ，α-SMAmRNA相对表达量比较
组别 n Co Ⅰ Col Ⅲ α-SMA 假手术组 9 0.25±0.03 0.13±0.04 0.31±0.03 模型组 8 0.57±0.08** 0.39±0.03** 0.57±0.06** 雷帕霉素组 9 0.35±0.05## 0.27±0.03## 0.44±0.05## 抗纤灵组 8 0.37±0.03## 0.28±0.03## 0.45±0.04## *P < 0.05 vs假手术组; **P < 0.01 vs假手术组; #P < 0.05 vs模型组; #P < 0.01 vs模型组.
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