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Abstract：
OBJECTIVE: Chronic renal failure (CRF) is a complex pathological condition that lacks a cure. Certain Chinese medicines, such as melittin, a major component in bee venom, have shown efficacy in treating CRF patients. On the other hand, the mechanisms underlying the therapeutic effects of melittin are unclear.
METHODS: A 5/6 nephrectomy model (5/6 Nx) of renal failure was established on rats for in vivo assays, and mouse podocyte clone 5 (MPC5) mouse podocyte cells were treated with angiotensin II (AngII) to establish an in vitro podocyte damage model. The 24-h urine protein, serum creatinine, and blood urea nitrogen levels were evaluated after one, 2, and 4 weeks. Hematoxylin and eosin staining, Masson staining, and periodic acid-Schiff staining were used to examine the pathological changes in kidney tissues. A cell counting kit 8 assay was used to assess the cell viability. Reverse transcription polymerase chain reaction and Western blot were used to assess the mRNA and protein levels in the cells, respectively.
RESULTS: In the rat 5/6 Nx, melittin reduced the 24-h urinary protein excretion and the serum creatinine and blood urea nitrogen levels. Furthermore, the renal pathology was improved in the melittin-treated 5/6 Nx rats. Melittin promoted podocin, nephrin, Beclin 1, and the LC3II/LC3I ratio and inhibited phosphorylated mammalian target of rapamycin (mTOR)/mTOR in 5/6 Nx-induced rats and AngII-induced MPC5 mouse podocyte cells. Moreover, inhibiting autophagy with 3-MA weakened the effects of melittin on podocin, nephrin, and the LC3II/LC3I ratio in podocytes.
CONCLUSIONS: Melittin may offer protection against kidney injury, probably by regulating podocyte autophagy. These results provide the theoretical basis for applying melittin in CRF therapy.
METHODS: A 5/6 nephrectomy model (5/6 Nx) of renal failure was established on rats for in vivo assays, and mouse podocyte clone 5 (MPC5) mouse podocyte cells were treated with angiotensin II (AngII) to establish an in vitro podocyte damage model. The 24-h urine protein, serum creatinine, and blood urea nitrogen levels were evaluated after one, 2, and 4 weeks. Hematoxylin and eosin staining, Masson staining, and periodic acid-Schiff staining were used to examine the pathological changes in kidney tissues. A cell counting kit 8 assay was used to assess the cell viability. Reverse transcription polymerase chain reaction and Western blot were used to assess the mRNA and protein levels in the cells, respectively.
RESULTS: In the rat 5/6 Nx, melittin reduced the 24-h urinary protein excretion and the serum creatinine and blood urea nitrogen levels. Furthermore, the renal pathology was improved in the melittin-treated 5/6 Nx rats. Melittin promoted podocin, nephrin, Beclin 1, and the LC3II/LC3I ratio and inhibited phosphorylated mammalian target of rapamycin (mTOR)/mTOR in 5/6 Nx-induced rats and AngII-induced MPC5 mouse podocyte cells. Moreover, inhibiting autophagy with 3-MA weakened the effects of melittin on podocin, nephrin, and the LC3II/LC3I ratio in podocytes.
CONCLUSIONS: Melittin may offer protection against kidney injury, probably by regulating podocyte autophagy. These results provide the theoretical basis for applying melittin in CRF therapy.
摘要：
目的：慢性肾功能衰竭（CRF）是一种复杂的病理状况，缺乏治愈方法。某些中药，比如蜂毒肽，蜂毒的主要成分，已显示出治疗CRF患者的疗效。另一方面,蜂毒素治疗作用的潜在机制尚不清楚.
方法：在大鼠体内建立5/6肾切除（5/6Nx）肾衰竭模型，用血管紧张素Ⅱ(AngII)处理小鼠足细胞克隆5(MPC5),建立体外足细胞损伤模型。24小时尿蛋白，血清肌酐,和血尿素氮水平进行评估后，2、4周。苏木精和伊红染色，Masson染色,高碘酸-希夫染色用于检查肾脏组织的病理变化。细胞计数试剂盒8测定用于评估细胞活力。逆转录聚合酶链反应和蛋白质印迹用于评估细胞中的mRNA和蛋白质水平，分别。
结果：在大鼠5/6Nx中，蜂毒素可降低24小时尿蛋白排泄以及血清肌酐和血尿素氮水平。此外，在蜂毒素治疗的5/6Nx大鼠中，肾脏病理学得到改善。蜂毒肽促进了波多辛，nephrin,Beclin1和LC3II/LC3I比例并抑制5/6Nx诱导的大鼠和AngII诱导的MPC5小鼠足细胞中雷帕霉素靶蛋白（mTOR）/mTOR的磷酸化。此外，用3-MA抑制自噬削弱了蜂毒素对podocin的影响，nephrin,和足细胞中的LC3II/LC3I比率。
结论：蜂毒肽可以提供对肾损伤的保护，可能是通过调节足细胞自噬。这些结果为蜂毒素在CRF治疗中的应用提供了理论依据。
方法：在大鼠体内建立5/6肾切除（5/6Nx）肾衰竭模型，用血管紧张素Ⅱ(AngII)处理小鼠足细胞克隆5(MPC5),建立体外足细胞损伤模型。24小时尿蛋白，血清肌酐,和血尿素氮水平进行评估后，2、4周。苏木精和伊红染色，Masson染色,高碘酸-希夫染色用于检查肾脏组织的病理变化。细胞计数试剂盒8测定用于评估细胞活力。逆转录聚合酶链反应和蛋白质印迹用于评估细胞中的mRNA和蛋白质水平，分别。
结果：在大鼠5/6Nx中，蜂毒素可降低24小时尿蛋白排泄以及血清肌酐和血尿素氮水平。此外，在蜂毒素治疗的5/6Nx大鼠中，肾脏病理学得到改善。蜂毒肽促进了波多辛，nephrin,Beclin1和LC3II/LC3I比例并抑制5/6Nx诱导的大鼠和AngII诱导的MPC5小鼠足细胞中雷帕霉素靶蛋白（mTOR）/mTOR的磷酸化。此外，用3-MA抑制自噬削弱了蜂毒素对podocin的影响，nephrin,和足细胞中的LC3II/LC3I比率。
结论：蜂毒肽可以提供对肾损伤的保护，可能是通过调节足细胞自噬。这些结果为蜂毒素在CRF治疗中的应用提供了理论依据。
