The susceptibility of cholesterol-depleted erythrocytes to saponin and sapogenin hemolysis

Segal, Ruth; Milo-Goldzweig, Ilana

doi:10.1016/0005-2736(78)90233-x

Cited by 35 publications

(16 citation statements)

References 13 publications

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“…This is similar to the result observed by Seebacher et al for an oleanolic acid aglycon. 12) In addition, the b configuration of the anomeric center between the two sugars (1, 2) is favored over the a one (3), and a terminal glucose (equatorial 4-OH) (1) slightly increases the activity as compared to a galactose (axial 4-OH) (2).…”

Section: )mentioning

confidence: 99%

“…1) This hemolytic activity has long been studied and the origin of the phenomenon is generally ascribed to their amphiphilic properties, but the exact mechanism is not yet known especially when considering the structural diversity of saponins. It has been shown that saponins are capable of interacting with membrane cholesterol [2][3][4] forming pits and holes which produce a destabilisation of the membrane by a micellar type-arrangement. 5) Some authors have postulated that the last step consists of an enzymatic deglycosylation that releases the aglycon and produces cell lysis.…”

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confidence: 99%

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Synthesis and Hemolytic Activity of Some Hederagenin Diglycosides

Chwalek

Plé

Voutquenne-Nazabadioko

2004

Chem. Pharm. Bull.

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Saponins are steroid or triterpenoid glycosides which are widespread in terrestrial and marine plants and possess various biological and pharmacological activities. 1) One of the first known activities of saponins is their ability to lyse erythrocytes.1) This hemolytic activity has long been studied and the origin of the phenomenon is generally ascribed to their amphiphilic properties, but the exact mechanism is not yet known especially when considering the structural diversity of saponins. It has been shown that saponins are capable of interacting with membrane cholesterol 2-4) forming pits and holes which produce a destabilisation of the membrane by a micellar type-arrangement. 5) Some authors have postulated that the last step consists of an enzymatic deglycosylation that releases the aglycon and produces cell lysis.6,7) Aglycons can show weak hemolytic activity but are not solely responsible for cell destruction. While the aglycon is very important in considering structure-activity relationships, the contribution of the sugar moiety is also significant. 8) Hemolytic activity depends on the nature, number and sequence of the sugars in the saponin.Our laboratory has long been interested in saponin isolation and identification, and, more recently, in their hemolytic structure-activity relationships. The study of hemolytic activity is a way of gaining insight into how saponins react at the membrane surface. We have compared many saponins with various triterpenoid aglycons and, as Schlösser and Wulff,8) have established that a polar substitution in ring A and weak polar substitution in rings D and E increases hemolytic activity and that monodesmosidic triterpenoid saponins are generally more active than bidesmosidic ones. 9)Structure-activity relationships of some synthetic saponins of oleanolic acid, 10-12) methyl oleanolate 13) and of other triterpenoid or steroid aglycons 14,15) have been previously established. It is thus known that the effect of the sugar residue on hemolytic activity is not transferable from one aglycon to one another. We wished to complete our study by analyzing the influence of the sugar units on hemolytic activity using hederagenin as the exclusive aglycon. Hederagenin saponins are largely represented in nature and possess many biological activities such as hemolytic, antiviral, antimicrobial, fongicidal, molluscicidal, or cytotoxic.1) With aglycons such as hederagenin, possessing a carboxylic group in position 28, it is known that hemolytic activity is greater when the free acid is esterified as a methyl ester. Unfortunately, no comparative structure-activity relationship have been performed on both the free acid and the methyl ester saponins to establish if the same factors systematically influence hemolytic activity. 8,16) We thus wished to synthesize the natural saponin 3hederagenin (isomaltosyl-hederagenin) 6 (Chart 1), and their corresponding methyl esters (1a, 2a, 3a, 4a, 5a, 6a). Compound 1 has been isolated from Fatsia japonica 17) and from the leaves of Barbarea vulgaris and ...

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Section: )mentioning

confidence: 99%

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confidence: 99%

Synthesis and Hemolytic Activity of Some Hederagenin Diglycosides

Chwalek

Plé

Voutquenne-Nazabadioko

2004

Chem. Pharm. Bull.

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“…If the OM of S. hyodysenteriae contained significant levels of cholesterol, as suggested by the extraction data, this membrane should be sensitive to agents that interact directly with sterols, such as the saponin digitonin. These compounds form a complex with cholesterol (20) and disrupt erythrocyte membranes (27,28). To see what effect digitonin had on S. hyodysenteriae, cells were incubated with various concentrations of digitonin.…”

Section: Resultsmentioning

confidence: 99%

Purified outer membranes of Serpulina hyodysenteriae contain cholesterol

et al. 1997

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We have isolated outer and inner membranes of Serpulina hyodysenteriae by using discontinuous sucrose density gradients. The outer and inner membrane fractions contained less than 1 and 2%, respectively, of the total NADH oxidase activity (soluble marker) in the cell lysate. Various membrane markers including lipooligosaccharide (LOS), the 16-kDa outer membrane lipoprotein (SmpA), and the C subunit of the F 1 F 0 ATPase indicated that the lowest-density membrane fraction contained outer membranes while the high-density membrane fraction contained inner membranes and that both are essentially free of contamination by the periplasmic flagella, a major contaminant of membranes isolated by other techniques. The outer membrane fractions ( ‫؍‬ 1.10 g/cm 3 ) contained 0. Swine dysentery, caused by Serpulina (formerly Treponema) hyodysenteriae (9, 10), is a highly infectious, acute to chronic disease that can result in a debilitating mucohemorrhagic diarrhea and death. The first stage of the disease is spirochetal colonization and proliferation in the enteric epithelium, resulting in a significant decrease in the absorption of nutrients, ions, and water (12). However, the spirochetes do not invade beyond the lamina propria, and putative virulence factors, such as hemolysins and lipooligosaccharide (LOS), probably cause the cellular damage (16,(21)(22)(23)(24)35). Because a close association of the bacteria with the intestinal lining is critical in the development of disease, surface components of the spirochetes must play a pivotal role in the process.Previously, different groups have attempted to isolate outer membranes (OMs) from S. hyodysenteriae with less than favorable results. This was mainly due to the contamination of OM fractions by periplasmic flagella. For example, Joens et al. (11) found seven major proteins, ranging from 42 to 32 kDa in size, in OMs isolated from S. hyodysenteriae B204 by a Sarkosyl extraction technique. However, six of the seven bands appeared to be derived from the periplasmic flagella. Similarly, Chatfield et al. (5) and Wannemuehler et al. (39) found that the majority of the putative OM proteins isolated with detergent ranged in size from 45 to 29 kDa and strongly resembled the protein profile observed for the flagellar proteins. These data suggested that membranes isolated by these techniques were heavily contaminated by flagella. Therefore, a more effective technique is needed to better define the OM of S. hyodysenteriae.We have developed a membrane separation technique that permits the isolation of S. hyodysenteriae OMs and inner membranes (IMs) with minimal contamination by flagella. Analysis of the OMs indicated that they are quite interesting. First, the OM of S. hyodysenteriae has a lower density than the IM, which is very unusual for bacteria. In addition, the OM contains significant levels of cholesterol. Previously, Mycoplasma species (6), Borrelia hermsii (15), and S. hyodysenteriae (30, 33) had been described as having sterols in their membranes; however, the distributions ...

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“…Unfortunately, the technique did not work as well with clinical specimens as it did with in vitro cultures. The mechanism of saponin resistance by P. falciparum -infected erythrocytes is unclear, but it is known that saponin binds to membrane cholesterol, and erythrocytes with decreased levels of membrane cholesterol were partially resistant to saponin destruction [16,17] .…”

Section: Discussionmentioning

confidence: 99%

Microscopic Detection of Mixed Malarial Infections: Improvement by Saponin Hemolysis

Orjih

Cherian²,

AlFadhli³

2008

Med Princ Pract

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Objective: The objective of the present study was to determine whether saponin hemolysis could improve microscopic detection of malaria parasites in human blood, since it has been previously reported that the technique has been used to enrich Plasmodium falciparum culture to ≧90% parasitemia. Material and Methods: Blood samples from suspected malaria cases were first examined in routine thick and thin smears under the microscope. The sample (1 ml) was then hemolyzed with 0.015% saponin in saline and centrifuged, the separated pellet was stained with Giemsa stain and examined microscopically, using PCR to confirm species identification. Results: With P. falciparum in vitro culture, the proportions of infected erythrocytes were approximately 0.7–2% before and 65–97% after saponin hemolysis, confirming published reports. In contrast, there was little or no increase in the proportions of intact infected erythrocytes after saponin hemolysis of clinical blood specimens. However, 20–600 hemolyzed parasites were detected per field under the microscope after saponin hemolysis of patients’ blood samples that contained only 1–15 parasites per field in conventional thick smears. In addition, more P. falciparum gametocytes were detected after saponin hemolysis. Conclusion: Saponin hemolysis concentrated the parasites in large volumes of blood into a small volume that could be smeared on a slide. This concentration method made it easy to detect malaria parasites and reduced the time needed for microscopy. In the present study, the method was comparable to PCR for the identification of P. vivax and P. falciparum mixed infections.

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The susceptibility of cholesterol-depleted erythrocytes to saponin and sapogenin hemolysis

Cited by 35 publications

References 13 publications

Synthesis and Hemolytic Activity of Some Hederagenin Diglycosides

Synthesis and Hemolytic Activity of Some Hederagenin Diglycosides

Purified outer membranes of Serpulina hyodysenteriae contain cholesterol

Microscopic Detection of Mixed Malarial Infections: Improvement by Saponin Hemolysis

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