Comparison of different extraction methods of exosome from macrophages / 中国药理学通报

Aim To establish the methods for exosomes isolation from the cell culture medium based on the principle of polyethy-lene glycol (PEG) precipitation combined with ultra-high speed centrifugation. Methods Exosomes were extracted by different centrifugation, PEG6000 precipitation combined with ultra-high speed centrifugation and EXO Quick kit,respectively. The mor-phology and particle size of exosomes were analyzed by transmis-sion electron microscopy,NanoSight ZS and Nano-particle track-er(NTA). Western blot assay was used to identify if the extrac-tion were exosomes. Results According to PEG6000 precipita-tion combined with ultra-high speed centrifugation method, the exosomes were successfully isolated from the culture medium. The morphology and particle size of exosomes,the exosomes iso-lated from PEG6000 precipitation combined with ultra-high speed centrifugation were equal to those of the ultra-high speed centrifugation but smaller to EXO Quick kit. What's more, the results of transmission electron microscopy showed that the three methods had typical discoid vesicles. Conclusion The im-provement of the exosome isolation from cell culture medium is convenient,easy and cheap.

Effect of compound Rhizoma Coptidis capsule on expression of transforming growth factor-beta1 and type IV collagen proteins in renal tissue of diabetic rats with nephropathy / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the effect and mechanism of compound Rhizoma Coptidis capsule (CRCC) on diabetic nephropathy in experimental rats.</p><p><b>METHOD</b>The rat model of early diabetic nephropathy was induced by injection of streptozotocin (STZ). The rats were divided into 6 groups: normal control group, model group, 3 CRCC treatment groups and XKW treatment group. The fasting blood glucose (FBG), blood urea nitrogen (BUN), creatinine (Cr), insulin (Ins) and urinary protein (Upro) were tested 30 days later. The expression of transforming growth factor-beta1 (TGF-beta1) and type IV collagen (IV-C) proteins and the pathological changes in renal tissue of diabetic rats with nephropathy were observed by optical micrography.</p><p><b>RESULT</b>CRCC could reduce the levels of FBG, BUN, Cr, Upro and the expression of TGF-beta1 and IV-C proteins, and alleviate pathological lesion in renal tissue of diabetic rats with nephropathy.</p><p><b>CONCLUSION</b>CRCC may protect the renal function and slow down the progression of diabetic nephropathy in rats by suppressing the expression of TGF-beta1 and IV-C proteins in renal tissue.</p>