Effects of aprotinin of organ cultures of the rat’s kidney

Davis, Heather L.; Gascho, C.; Kiernan, J. A.

doi:10.1007/bf02796441

“…Aprotinin is a well-characterized serine protease inhibitor extracted from bovine lung and known not to penetrate the cell membrane (Fritz and Wunderer, 1983;Hewlett, 1990). Addition of aprotinin improved cell survival in rat renal tissue cultures by protecting membrane-bound glycoproteins from proteolytic enzymes (Davis et al, 1976). Aprotinin has also been found to increase the survival of rat hepatocytes presumably by inhibiting serine proteases located in the plasma membrane Nakamura et al, 1984) and to promote growth of transformed rat embryo fibroblasts, an effect attributed to decreased proteolysis of autocrine growth factors (Cook and Chen, 1988).…”

Section: Discussionmentioning

confidence: 98%

Protease Activity in Protein-Free Ns0 Myeloma Cell Cultures

Spens

¹

,

Häggström

²

2005

In Vitro Cell Dev Biol Anim

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Zymography of concentrated conditioned medium (CM) from protein-free NS0 myeloma cell cultures showed that this cell line produced and released/secreted several proteases. Two caseinolytic activities at 45-50 and 90 kDa were identified as aspartic acid proteases, and at least two cathepsins of the papain-like cysteine protease family with molecular masses of 30-35 kDa were found by gelatin zymography. One of these cathepsins was identified as cathepsin L by using an enzyme assay exploiting the substrate Z-Phe-Arg-AMC and the inhibitor Z-Phe-Tyr-t(Bu)-DMK. The aspartic acid and cysteine proteases were active only at acidic pH and are therefore not a potential risk for degrading the product or affecting cell growth during culture. Secreted proforms of cathepsins may, however, possess mitogenic functions, but addition of anti-procathepsin L antibodies to NS0 cultures did not influence proliferation. The recombinant antibody product was not degraded in cell-free CM incubated at pH 7, but when the pH was decreased to 3.5-4, the aspartic acid proteases degraded the product. Gelatin zymography also revealed the presence of several serine proteases in NS0 CM, one at 85 kDa and two at 50 kDa, with pH optima close to culture pH. Addition of the serine protease inhibitor aprotinin significantly increased the specific proliferation rate as compared to the control. In addition to these data, N-terminal amino acid sequencing identified two proteins in NS0 CM as the protease inhibitors secretory leukocyte protease inhibitor and cystatin C.

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“…Aprotinin exerts various effects on tissue when used in a tissue culture model at extremely high concentrations (up to 10 7 KIU/L). 22,23 The effect of aprotinin on organ preservation-liver, kidney, and lung-has also been studied. [24][25][26][27][28][29] This proteinase inhibitor was shown to protect dog myocardium against ischemic injury in terms of diminished area of infarction after coronary artery ligation.…”

Section: Discussionmentioning

confidence: 99%

Aprotinin improves myocardial recovery after ischemia and reperfusion:

Gurevitch¹,

Barak²,

Hochhauser³

et al. 1994

The Journal of Thoracic and Cardiovascular Surgery

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Aprotinin improves myocardial recovery after ischemia and reperfusion Effects of the drug on isolated rat heartsThe effects of aprotinin, a protease inhibitor, on the ischemic and nonischemic isolated rat heart was investigated with the use of the modified Langendorlf model. During phase I of the study, hearts were perfused with either low-dose aprotinin (10 5 Kill/L), high-dose aprotinin (10 6 KIU /L), or normal saline solution added to modified Krebs-Henseleit solution. No statistically significant differences in contraction amplitude, contractility, coronary flow, and wet/dry heart weight ratio were observed among the three groups of hearts. In phase II, hearts were exposed to a 40-minute period of global ischemia at 31 0 C. Ischemic arrest was induced by warm cardioplegia. Before ischemia and during cardioplegia, hearts were perfused with either aprotinin 10 6 KIU/L (n = 10) or normal saline solution (n = 10) for .30 minutes. On reperfusion, recovery of hearts treated with aprotinin was significantly better than that of control hearts, as reflected by better contractility (analysis of variance, p = 0.011), higher coronary flow (p < 0.025), and lower creatine kinase levels (p <0.05). No statistically significant differences in contraction amplitude were observed between the two groups. When the effect of ischemia within each group of hearts was analyzed, the preserving effect of aprotinin was even more pronounced. In the control group, ischemia caused a decrease in contractility (p < 0.025) and a decrease in oxygen consumption (p = 0.006); by contrast, in the aprotinin group the preischemic values were maintained. Accordingly, we conclude that aprotinin at concentrations up to 10 6 KIU /L has no deleterious effect on normally perfused hearts and has a significant protective effect on the ischemic heart when used in high doses in the preischemic period. (J THoRAe CARDIOVASC SURG 1994;108:109-18)

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“…The pH profile suggests that serine protease activity is present in the culture at pH 7.15 (Fig. Addition of aprotinin improved cell survival in rat renal tissue cultures hy protecting membrane-bound glyeoproteins from proteolytie enzymes (Davis et al, 1976). As judged by the growth-promoting eft~et of added aprotinin, the serine protease activity appears to inhibit NS0 cell proliferation (Fig.…”

Section: Discussionmentioning

confidence: 98%

Protease activity in protein-free NSO myeloma cell cultures

Spens

¹

,

Häggström

²

2005

In Vitro Cell.Dev.Biol.-Animal

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Zymography of concentrated conditioned medium (CM) from protein-free NS0 myeloma cell cultures showed that this cell line produced and released/secreted several proteases. Two caseinolytic activities at 45-50 and 90 kDa were identified as aspartic acid proteases, and at least two cathepsins of the papain-like cysteine protease family with molecular masses of 30-35 kDa were found by gelatin zymography. One of these cathepsins was identified as cathepsin L by using an enzyme assay exploiting the substrate Z-Phe-Arg-AMC and the inhibitor Z-Phe-Tyr-t(Bu)-DMK. The aspartic acid and cysteine proteases were active only at acidic pH and are therefore not a potential risk for degrading the product or affecting cell growth during culture. Secreted proforms of cathepsins may, however, possess mitogenic functions, but addition of anti-procathepsin L antibodies to NS0 cultures did not influence proliferation. The recombinant antibody product was not degraded in cell-free CM incubated at pH 7, but when the pH was decreased to 3.5-4, the aspartic acid proteases degraded the product. Gelatin zymography also revealed the presence of several serine proteases in NS0 CM, one at 85 kDa and two at 50 kDa, with pH optima close to culture pH. Addition of the serine protease inhibitor aprotinin significantly increased the specific proliferation rate as compared to the control. In addition to these data, N-terminal amino acid sequencing identified two proteins in NS0 CM as the protease inhibitors secretory leukocyte protease inhibitor and cystatin C.

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Effects of aprotinin of organ cultures of the rat’s kidney

Cited by 14 publications

References 8 publications

Protease Activity in Protein-Free Ns0 Myeloma Cell Cultures

Protease Activity in Protein-Free Ns0 Myeloma Cell Cultures

Aprotinin improves myocardial recovery after ischemia and reperfusion:

Protease activity in protein-free NSO myeloma cell cultures

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