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排毒保肾丸对5/6肾切除大鼠肾纤维化的影响

[Effect of Paidu Baoshen Pill on renal fibrosis in 5/6 nephrectomized rats].

作者信息

Wang Shui-Hua, Chen Bang-Ming, Liu Yong-Fang, Che Wei-Ping, Wu Zhao-Dong, Wang Guo-Bing, Xia Xiao-Qin, Huang Hong-En, Wei Lin, Zhu Hai-Long, Liu Gan-Yan

出版信息

Zhongguo Zhong Xi Yi Jie He Za Zhi. 2015 Jan;35(1):81-7.

PMID:25790680
Abstract

OBJECTIVE

To observe the anti-renal fibrosis effect of Paidu Baoshen Pill (PBP) on 5/6 nephrectomized rats and to explore its mechanism.

METHODS

Totally 50 SD male healthy rats were randomly divided into the normal control group (n = 10), the sham-operation group (n = 10), and the nephrectomy model group (n = 30) according to the proportion of 1:1:3. Rats in the sham-operation group had their renal capsule isolated without nephrectomy. Rats in the nephrectomy model group had their kidneys 5/6 nephrectomized. Then 24 h urine was collected and 24 h urinary protein (24 h UP) detected. Serum blood urea nitrogen (BUN) and serum creatitine (SCr) were also tested. According to the SCr level 30 rats of the model group were further randomly divided into the model group, the PBP group, and the Niaoduqing Granule (NG) group, 10 in each group. Rats in the PBP group and the NG group were respectively administered with PBP (at the daily dose of 1.0 g/kg) and NG (at the daily dose of 3.33 g/kg) by gastrogavage (they were dissolved in distilled water). At the same time, 2 mL distilled water was administered by gastrogavage to rats in the normal control group, the sham-operation group, and the nephrectomy model group, once daily for 4 successive weeks. Mental conditions, activities, hair color, shape of stool, and the body weight were observed during administration. After 4 weeks, urine was collected to detect 24 h UP. Blood was sampled to detect SCr, BUN, transforming growth factor β1 (TGF-β1), type III procollagen (PC III), collagen type IV (Col IV), laminin (LN), and fibronectin (FN). After rats were killed, their left remnant renal tissues were collected for pathological examinations. The protein expression quantity of TGF-β1 and FN was detected by immunohistochemical method. mRNA expression levels of TGF-β1 and FN were detected using real time fluorescent quantitative PCR.

RESULTS

There was no statistical difference in the above indices between the normal control group and the sham-operation group (P > 0.05). Compared with the sham-operation group, rats' general condition was poorer in the model group, their body weight grew slower, and 24 h UP increased; serum levels of BUN, SCr, TGF-β1, PC III, Col IV, LN, and FN increased; the residual renal pathological lesion was serious; expression levels of TGF-β1, TGF-β1, mRNA, FN, and FN mRNA increased in the renal tissue (all P < 0.01). Compared with the model group, rats' general condition was better, their body weight grew faster, 24 h UP reduced (P < 0.05), blood levels of BUN and SCr decreased significantly (P < 0.01), serum levels of TGF-β1, PC III, CoL IV, LN, and FN decreased (P < 0.05, P < 0.01); the residual renal pathological lesion was attenuated in the PBP group and the NG group; expression levels of TGF-β1, TGF-β1, mRNA, FN, and FN mRNA decreased (P < 0.01). Compared with the NG group, blood levels of SCr and FN, and expression levels of FN and FN mRNA decreased more in the PBP group (P < 0.05).

CONCLUSIONS

PBP had the effect of anti-renal fibro- sis in 5/6 nephrectomized rats. Down-regulating expression levels of TGF-β1, and FN from gene transcription and protein translation levels might be one of its mechanisms.

摘要

目的

观察排毒保肾丸(PBP)对5/6肾切除大鼠抗肾纤维化的作用并探讨其机制。

方法

将50只健康雄性SD大鼠按1∶1∶3比例随机分为正常对照组(n = 10)、假手术组(n = 10)和肾切除模型组(n = 30)。假手术组大鼠仅分离肾包膜不切除肾脏;肾切除模型组大鼠行5/6肾切除术。术后收集24 h尿液检测24 h尿蛋白(24 h UP),同时检测血清尿素氮(BUN)和血清肌酐(SCr)。根据SCr水平将模型组30只大鼠再随机分为模型组、PBP组和尿毒清颗粒(NG)组,每组10只。PBP组和NG组大鼠分别按1.0 g/kg、3.33 g/kg的剂量灌胃给予PBP和NG(均用蒸馏水溶解),正常对照组、假手术组和肾切除模型组大鼠同时灌胃给予2 mL蒸馏水,每日1次,连续4周。给药期间观察大鼠精神状态、活动情况、毛色、粪便形状及体质量变化。4周后收集尿液检测24 h UP,取血检测SCr、BUN、转化生长因子β1(TGF-β1)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(ColⅣ)、层粘连蛋白(LN)及纤维连接蛋白(FN)。处死大鼠后取左肾残留组织进行病理检查,采用免疫组化法检测TGF-β1和FN的蛋白表达量,采用实时荧光定量PCR检测TGF-β1和FN的mRNA表达水平。

结果

正常对照组与假手术组上述指标比较差异无统计学意义(P>0.05)。与假手术组比较,模型组大鼠一般状况较差,体质量增长缓慢,24 h UP升高;血清BUN、SCr、TGF-β1、PCⅢ、ColⅣ、LN及FN水平升高;残留肾病理损伤严重;肾组织中TGF-β1、TGF-β1 mRNA、FN及FN mRNA表达水平升高(均P<0.01)。与模型组比较,PBP组和NG组大鼠一般状况较好,体质量增长较快,24 h UP降低(P<0.05),血BUN和SCr水平显著降低(P<0.01),血清TGF-β1、PCⅢ、ColⅣ、LN及FN水平降低(P<0.05,P<0.01);残留肾病理损伤减轻;肾组织中TGF-β1、TGF-β1 mRNA、FN及FN mRNA表达水平降低(P<0.01)。与NG组比较,PBP组血SCr和FN水平及FN和FN mRNA表达水平降低更明显(P<0.05)。

结论

PBP对5/6肾切除大鼠具有抗肾纤维化作用,从基因转录和蛋白翻译水平下调TGF-β1和FN表达水平可能是其作用机制之一。

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