Effect of Biling Qutong Prescription on SIRT3 Protein Expression and URAT1 mRNA in Skeletal Muscle of Diabetic Gout Rats

Zhong-nan LI; Yan-yang XING; Yuan-yuan ZHOU; Shu-hong MA; Yu-ting XING; De-mei DOU; Zhao-hui FANG

doi:10.13422/j.cnki.syfjx.20191903

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Effect of Biling Qutong Prescription on SIRT3 Protein Expression and URAT1 mRNA in Skeletal Muscle of Diabetic Gout Rats

Pharmacology | 更新时间：2021-02-09

- Effect of Biling Qutong Prescription on SIRT3 Protein Expression and URAT1 mRNA in Skeletal Muscle of Diabetic Gout Rats
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 21, Pages: 25-31(2019)
- 作者机构：
  
  1.安徽中医药大学　第一附属医院，合肥　 230031；
  2.安徽中医药大学　研究生院，合肥　 230038
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20191903
  CLC： R2-0; R22; R285.5; R289
- Published：05 November 2019，
  
  Published Online：19 June 2019，
  
  Received：18 February 2019，
- 稿件说明：
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Zhong-nan LI, Yan-yang XING, Yuan-yuan ZHOU, et al. Effect of Biling Qutong Prescription on SIRT3 Protein Expression and URAT1 mRNA in Skeletal Muscle of Diabetic Gout Rats. [J]. Chinese Journal of Experimental Traditional Medical Formulae 25(21):25-31(2019)
DOI：

Zhong-nan LI, Yan-yang XING, Yuan-yuan ZHOU, et al. Effect of Biling Qutong Prescription on SIRT3 Protein Expression and URAT1 mRNA in Skeletal Muscle of Diabetic Gout Rats. [J]. Chinese Journal of Experimental Traditional Medical Formulae 25(21):25-31(2019) DOI： 10.13422/j.cnki.syfjx.20191903.

摘要

目的：

探讨萆苓祛痛方对糖尿病痛风大鼠骨骼肌组织去乙酰化酶3(SIRT3)蛋白表达及尿酸盐转运体1(URAT1) mRNA的影响。

方法：

选择健康雄性大鼠40只，除正常组外，其余组予高脂饲料喂养并联合小剂量链脲佐菌素(STZ)溶液40 mg·kg

－1

腹腔注射1次，以血糖≥16.7 mmol·L

－1

，为糖尿病模型。4 d后关节腔注射5%尿酸钠溶液1次，诱导痛风模型，模型成功后，分为萆苓祛痛方组(萆苓组，10 g·kg

－1

)，吲哚美辛组(5 mg·kg

－1

)，吡格列酮组(10 mg·kg

－1

)，均连续给药21 d，正常组、模型组予等量生理盐水；采用蛋白免疫印迹法(Western blot)测定骨骼肌组织SIRT3蛋白表达；实时荧光定量聚合酶链式反应(Real-time PCR)检测骨骼肌组织URAT1 mRNA表达，并进行病理检查，取血测定血糖(GLU)，血尿酸(UA)及C反应蛋白(CRP)含量。

结果：

与正常组比较，模型组GLU，UA及CRP明显升高(

0.01)；与模型组比较，萆苓组、吡格列酮组血糖下降(

0.05)；各药物组UA及CRP明显下降(

0.01)。与正常组比较，模型组骨骼肌SIRT3蛋白表达量显著降低(

0.01)；与模型组比较，萆苓组骨骼肌SIRT3蛋白表达量显著提高(

0.01)，与西药组比较无明显差异；条带图的结果同样显示，与正常组比较，模型组表达亮度明显减弱，药物组表达亮度明显增强；与正常组比较，模型组关节组织URAT1 mRNA相对表达量明显升高(

0.01)；与模型组比较，各药物组URAT1 mRNA相对表达量显著下调(

0.01)。电泳图同样提示，正常组表达亮度减弱，模型组表达亮度显著增强，萆苓组、西药组表达亮度明显减弱。关节病理提示，与正常组比较，模型组大鼠关节病理损伤严重，可见大量炎细胞浸润及纤维增生，滑膜细胞变性、坏死。与模型组比较，萆苓祛痛方关节病变程度明显减低，见少量炎细胞浸润，滑膜上皮轻度增生。

结论：

具有泻浊解毒通络作用的萆苓祛痛方可显著提高糖尿病痛风大鼠骨骼肌组织SIRT3的蛋白表达量，下调URAT1 mRNA的表达量，减轻骨骼肌组织病理损伤，减低血清炎症因子CRP的含量，降低模型大鼠的血糖、血尿酸水平，有保护关节功能的作用。

Abstract

Objective:

To observe the effect of phlegm and blood stasis on the expressions of sirtuin 3(SIRT3)protein and urate transporter 1(URAT1) mRNA in skeletal muscle of diabetic rats with gout.

Method:

The 40 healthy rats

excepting the normal group

the remaining groups were fed with high-fat diet combined with low-dose streptozotocin solution (40 mg·kg

-1

) once a day

with blood glucose "16.7 mmol·L

-1

" as the criterion for the diabetes model. After 4 days

the 5%sodium urate solution was injected into the joint cavity once to induce the gout model. After the successful modeling

the Biling group (10 g·kg

-1

)

the indomethacin group (5 mg·kg

-1

) and the pioglitazone group (10 mg·kg

-1

) continued to be administered for 21 days. The normal group and the model group were given the same amount of normal saline. The expression of SIRT3 protein in skeletal muscle tissue was determined by Western blot

URAT1 mRNA expression in bone tissue was detected by quantitative real-time fluorescence polymerase chain reaction(Real-time PCR)

and blood was collected to measure blood glucose (GLU)

blood uric acid (UA) and C-reactive protein (CRP).

Result:

Compared with the normal group

GLU

UA and CRP in the model group were significantly increased (

0.01). Compared with the model group

GLU in the Biling group and the pioglitazone group were decreased

and UA and CRP in the medicine group were significantly decreased (

0.05

0.01). Compared with the normal group

the expression of SIRT3 protein in the skeletal muscle of the model group was significantly decreased (

0.01)

while the expression of SIRT3 protein in the skeletal muscle of the Biling group was significantly increased after administration (

0.01). Compared with the model group

the expression of SIRT3 protein in the skeletal muscle of the sputum group was significantly increased (

0.01)

with no significant difference from the western medicine group. The results of the strip chart also showed that compared with the model group

the expression brightness of the model group was significantly weakened

while the expression brightness of the drug group was significantly enhanced. Compared with the normal group

the relative expression of joint URAT1 mRNA in the model group was significantly increased (

0.01). Compared with the model group

the relative expression of URAT1 mRNA in each drug group was significantly down-regulated (

0.01). The results of the strip chart also showed that expression brightness of the normal group was the weakest

while that of the model group was the highest

and the expression brightness of the Biling group and the western medicine group was significantly weakened. Joint pathology suggested that compared with the normal group

the pathological damage of the joints in the model group was severe

with a large number of inflammatory cell infiltration and fibrosis

synovial cell degeneration and necrosis. Compared with the model group

the degree of joint disease was significantly reduced after treatment with Biling Qutong prescription

with only a small amount of inflammatory cell infiltration and mild hyperplasia in synovial epithelium.

Conclusion:

Biling Qutong prescription with effects in purging turbidity

detoxifying and dredging collaterals can significantly reduce the content of serum inflammatory factor CRP

significantly increase the protein expression of SIRT3 in skeletal muscle tissue of model rats

lower the content of URAT1 mRNA

reduce the blood glucose and blood uric acid levels in diabetic gout rats

and protect joints.

关键词

萆苓祛痛方糖尿病痛风大鼠骨骼肌组织去乙酰化酶3(SIRT3)尿酸盐转运体1(URAT1)

Keywords

Biling Qutong prescriptiondiabetic gout ratsskeletal muscle tissuesirtuin 3(SIRT3)urate transporter 1 (URAT1)

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