Ao Qiangguo, Cheng Qingli, Ma Qiang, Wang Xiaodan, Liu Sheng
Department of Geriatric Nephrology, Institute of Gerontology, Chinese PLA General Hospital, China.
Intern Med. 2013;52(10):1035-41. doi: 10.2169/internalmedicine.52.9574. Epub 2013 May 15.
Evidence has demonstrated that Ca(2+)/calmodulin-dependent protein kinase type IV (CaMKIV) contributes to altered cytokine production by promoting the production of inflammatory cytokines. This study aimed to explore the protective role and underlying mechanisms of CaMKIV inhibition in experimental nephrotic syndrome.
BALB/c mice received single intravenous injections of adriamycin (10 mg/kg) then were sacrificed at two, four and six weeks. In the second study, treatment with KN-93, a CaMKIV inhibitor, or vehicle administered via intraperitoneal injection was started five days after adriamycin injection. Functional and pathologic parameters, the presence of inflammatory infiltration and the expressions of pro-inflammatory cytokines were assessed.
The CaMKIV protein expression levels were upregulated in the mice with adriamycin nephropathy, which was significantly inhibited by KN-93 (p<0.01). As compared with the vehicle-treated controls, KN-93 treatment resulted in marked suppression of proteinuria and serum creatinine at week 6 (p<0.01), but not at two weeks after induction of the disease. KN-93 inhibited glomerulosclerosis and the development of tubulointerstitial lesions. The renal alpha-smooth muscle actin (α-SMA) expression was also significantly suppressed by KN-93 treatment at week 6 (p<0.01). Moreover, KN-93 inhibited the renal monocyte chemoattractant protein-1 (MCP-1) expression, paralleled by a reduction in the interstitial infiltration of macrophages and T-cells (p<0.01).
Our findings suggest that activation of CaMKIV signaling is involved in the progression of glomerular diseases with a proteinuric state. Our data therefore justify the development of small molecule CaMKIV inhibitors for the treatment of clinical nephrotic syndrome.
有证据表明,Ⅳ型钙/钙调蛋白依赖性蛋白激酶(CaMKIV)通过促进炎性细胞因子的产生,导致细胞因子产生改变。本研究旨在探讨抑制CaMKIV在实验性肾病综合征中的保护作用及潜在机制。
BALB/c小鼠单次静脉注射阿霉素(10 mg/kg),然后在2周、4周和6周时处死。在第二项研究中,在阿霉素注射5天后开始通过腹腔注射给予CaMKIV抑制剂KN-93或赋形剂。评估功能和病理参数、炎性浸润情况以及促炎细胞因子的表达。
阿霉素肾病小鼠中CaMKIV蛋白表达水平上调,KN-93可显著抑制该上调(p<0.01)。与赋形剂处理的对照组相比,KN-93处理在第6周时可显著抑制蛋白尿和血清肌酐水平(p<0.01),但在疾病诱导后2周时无此作用。KN-93抑制肾小球硬化和肾小管间质病变的发展。在第6周时,KN-93处理还可显著抑制肾α-平滑肌肌动蛋白(α-SMA)的表达(p<0.01)。此外,KN-93抑制肾单核细胞趋化蛋白-1(MCP-1)的表达,同时巨噬细胞和T细胞的间质浸润减少(p<0.01)。
我们的研究结果表明,CaMKIV信号通路的激活参与了伴有蛋白尿状态的肾小球疾病的进展。因此,我们的数据证明开发小分子CaMKIV抑制剂用于治疗临床肾病综合征是合理的。