补肾活血方对慢性肾衰竭大鼠氧化应激、PI3K/Akt、Bcl-2/Bax的影响

Effect of Bushen Huoxue Prescription on Oxidative Stress,PI3K/Akt,Bcl-2/Bax in Rats with Chronic Renal Failure

【目的】探讨补肾活血方对慢性肾衰竭大鼠的治疗作用及机制。【方法】将40只SD大鼠分为正常组(10只)、模型组(9只)、补肾活血方低剂量组(10只)和补肾活血方高剂量组(10只)。除正常组外,其他组别大鼠采用腺嘌呤灌胃法复制慢性肾衰竭模型。造模成功后,补肾活血方低、高剂量组大鼠分别给予0.52、2.08 g/kg补肾活血方药液灌胃,每日1次,连续给药28 d。给药结束后,酶联免疫吸附法(ELISA)检测血清肾功能指标尿素氮(BUN)、血肌酐(SCr)和尿酸(UA)水平及肾组织氧化应激指标丙二醛(MDA)、超氧化物歧化酶(SOD)水平,苏木素-伊红(HE)染色法观察肾组织病理学变化,Western Blot法检测肾组织磷脂酰肌醇-3激酶(PI3K)、蛋白激酶B(Akt)、B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)蛋白表达情况。【结果】(1)与正常组比较,模型组BUN、SCr、UA水平均上升(P<0.05);与模型组比较,补肾活血方低、高剂量组BUN、SCr、UA水平均显著下降(P<0.05)。(2)与正常组比较,模型组MAD水平上升,SOD水平下降(P<0.05);与模型组比较,补肾活血方低、高剂量组MAD水平下降,SOD水平上升(P<0.05);(3)与正常组比较,模型组肾组织中出现轻度坏死和炎性细胞浸润,肾小管扩张,肾小球变形等病理学变化;与模型组比较,补肾活血方低、高剂量组肾组织病理情况均有改善。(4)与正常组比较,模型组大鼠肾组织PI3K、Akt蛋白表达水平显著升高(P<0.05);与模型组比较,补肾活血方低、高剂量组大鼠肾组织PI3K、Akt蛋白表达水平显著降低(P<0.05)。(5)与正常组比较,模型组大鼠肾组织Bcl-2蛋白表达水平下降,Bax蛋白表达水平上升(P<0.05);与模型组比较,补肾活血方低、高剂量组大鼠肾组织Bcl-2蛋白表达水平上升,Bax蛋白表达水平下降(P<0.05)。【结论】补肾活血方能够改善慢性肾衰竭大鼠肾脏损伤,其机制与减轻氧化应激、抑制PI3K/Akt信号通路、调控凋亡蛋白Bcl-2/Bax表达有关。

Objective To investigate the therapeutic effect and mechanism of the Bushen Huoxue Prescription(BHP, a herbal compound mainly with the actions of replenishing kidney and activating blood)in rats with chronic renal failure. Methods Forty SD rats were divided into normal group(10 rats),model group(9 rats),BHP low-dose group(10 rats)and high-dose group(10 rats). Except for the normal group,the rats in the all groups were replicated with the chronic renal failure model by adenine gavage. After successful replication,the rats in the BHP low-and high-dose groups were given 0.52 and 2.08 g/kg of BHP by gavage,respectively,once daily for28 consecutive days. After the end of administration,serum levels of renal function indicators blood urea nitrogen(BUN), blood creatinine(SCr)and uric acid(UA), and levels of oxidative stress indexes malondialdehyde(MDA)and superoxide dismutase(SOD)in renal tissue were measured by enzyme-linked immunosorbent assay(ELISA),the histopathological changes of rat kidney tissue were observed by hematoxylin-eosin(HE)staining,and the protein expressions of phosphatidylinositol-3-kinase(PI3K),protein kinase B(Akt),B cell lymphoma 2(Bcl-2)and Bcl-2 associated X protein(Bax)in renal tissue were detected by Western Blot. Results(1)Compared with the normal group,the levels of BUN,SCr and UA were increased in the model group(P<0.05);compared with the model group,the levels of BUN,SCr and UA were decreased significantly in the BHP low-and highdose groups(P<0.05).(2)Compared with the normal group, MAD level was increased and SOD level was decreased in the model group(P<0.05);compared with the model group,MAD level was decreased and SOD level increased in the BHP low-and high-dose groups(P<0.05);(3)Compared with the normal group,pathological changes such as mild necrosis and inflammatory cell infiltration, tubular dilatation and glomerular deformation appeared in the kidney tissue of the model group;compared with the model group,the pathological conditions in both the BHP low-and high-dose groups were improved.(4)Compared with the normal group,the protein expression levels of PI3K and Akt in the kidney tissue in the model group were significantly increased(P<0.05);compared with the model group,the protein expression levels of PI3K and Akt in the kidney tissue in the BHP low-and high-dose groups were significantly decreased(P<0.05).(5)Compared with the normal group,the protein expression level of Bcl-2 in the kidney tissue in the model group was decreased and the protein expression level of Bax was increased(P<0.05);compared with the model group,the expression level of Bcl-2protein in the kidney tissue in the BHP low-and high-dose groups was increased and the protein expression level of Bax was decreased(P<0.05). Conclusion BHP has effect in improving kidney injury in rats with chronic renal failure by a mechanism related to inhibition of PI3K/Akt signaling pathway and regulation of related apoptotic protein expression.