Objectives

In the present study, we aimed to assess whether adrenocorticotropic hormone (ACTH) could protect the podocytes from adriamycin (ADR)-induced injury by stimulating B lymphocytes to secrete the associated cytokines.

Methods

Proliferation assay was used to assess the proliferation and activity of podocytes. Enzyme-linked immunosorbent assay was used to examine the secretion of IL-10 and IL-4. TUNEL apoptosis detection kit was used to detect the apoptosis of podocytes. Real-time PCR and Western blotting analysis were used to examine the expressions of nephrin and podocin at the mRNA and protein levels.

Results

Compared with the normal control group, the podocyte proliferation of ADR group was significantly inhibited. However, compared with the ADR group, the podocyte proliferation of the supernatant (1 µg/L, 10 µg/L or 100 µg/L ACTH_4-10) + ADR groups was generally increased, and the pro-proliferative effect of the supernatant containing 10 µg/L ACTH_4-10 was the highest. Moreover, we found that after B lymphocytes were intervened by 10 µg/L ACTH4-10, the IL-10 level in the cell supernatant was significantly elevated (p < 0.05). When anti-IL-10R was added, the podocyte proliferation of the supernatant (10 µg/L ACTH_4-10) + ADR group was significantly inhibited. Furthermore, the supernatant of B cells stimulated with 10 µg/L ACTH_4-10 could better decrease the apoptosis rate of injured podocytes and increase the expressions of nephrin and podocin at the mRNA and protein levels by elevating the secretion of IL-10.

Conclusion

Compared with ACTH_4-10, the supernatant of B cells stimulated with ACTH_4-10 could better protect the podocytes from ADR-induced injury by elevating the secretion of IL-10.

中文翻译：

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ACTH4-10通过刺激B淋巴细胞分泌白介素10来保护ADR损伤的足细胞。

目标

在本研究中，我们旨在评估促肾上腺皮质激素（ACTH）是否可以通过刺激B淋巴细胞分泌相关的细胞因子来保护足细胞免受阿霉素（ADR）诱导的损伤。

方法

增殖测定用于评估足细胞的增殖和活性。酶联免疫吸附试验用于检查IL-10和IL-4的分泌。TUNEL凋亡检测试剂盒用于检测足细胞的凋亡。实时PCR和Western印迹分析用于检查nephrin和podocin在mRNA和蛋白质水平的表达。

结果

与正常对照组相比，ADR组足细胞增殖受到明显抑制。但是，与ADR组相比，上清液（1 µg / L，10 µg / L或100 µg / L ACTH _4-10）+ ADR组的足细胞增殖普遍增加，并且上清液具有促增殖作用含10 µg / L ACTH _4-10的含量最高。此外，我们发现在B淋巴细胞受到10 µg / L ACTH4-10的干预后，细胞上清液中的IL-10水平显着升高（p <0.05）。加入抗IL-10R后，上清液（10 µg / L ACTH _4-10）+ ADR组的足细胞增殖受到明显抑制。此外，用10 µg / L ACTH _4-10刺激的B细胞上清液 通过增加IL-10的分泌，可以更好地降低受损足细胞的凋亡率，并增加nephrin和podocin在mRNA和蛋白质水平上的表达。

结论

用ACTH比较_4-10， B细胞的上清液用ACTH刺激_4-10可以更好地通过提高IL-10的分泌保护ADR引起的损伤足细胞。