The Acute Encephalopathy Induced by Intake of Sugihiratake Mushroom in the Patients with Renal Damage Might Be Associated with the Intoxication of Cyanide and Thiocyanate

Abstract

A novel type of encephalopathy associated with the ingestion of Sugihiratake mushroom (Pleurocybella porrigens) occurred in patients with chronic renal failure treated on hemodialysis in fall, 2004 in Japan. To clarify the mechanism of encephalopathy onset, we, for the first time, purified the cyanogen glycoside fraction (CG) from Sugihiratake mushroom using reversed phase high-performance liquid chromatography and hydrophilic interaction chromatography. Furthermore, we investigated single dose toxicity of the CG in an adenine-induced rat model of chronic renal damage (CRD). Pathological examination of kidneys indicates the development of CRD. Oral administration of the CG induces the accumulation of thiocyanate in the hemolyzed blood and brain in CRD rats, although no morphological changes were found in the brain. No further enhancement of kidney damage is observed after the oral administration of the CG in CRD rats. This is the first experimental report to suggest that acute encephalopathy, induced by Sugihiratake mushroom intake in the patients with chronic renal failure, is associated with intoxication of cyanide and thiocyanate, presumably produced metabolically produced after the ingestion of Sugihiratake mushroom.

Figures

Fig. 1.

Cyanide (%) content of fractions obtained from hydrophilic interaction chromatography.

The vertical axis represents the ratio of cyanide content detected in each fraction to total cyanide content detected in the sample before the injection to HPLC.

Fig. 2.

Temporal changes of cyanide levels in the hemolyzed blood in an adenine-induced rat model of CRD.

Control represents the control group administered distilled water alone, Low-dose represents the low-dose group administered 360 mg cyanogen glycoside fraction (CG)/kg body weight (corresponding to 0.31 mg sodium cyanide/kg body weight), and High-dose represents the high-dose group administered 2840 mg CG/kg body weight (corresponding to 2.44 mg sodium cyanide/kg body weight). Bars represent means ± standard deviation (3 rats/group).

The bar represents the change of cyanide level in the hemolyzed blood from 0 h to 8 h after administration of CG

The bar represents the change of cyanide level in the hemolyzed blood from 0 h to 24 h after administration of CG

Fig. 3.

Temporal changes of thiocyanate levels in the hemolyzed blood in an adenine-induced rat model of CRD.

Control represents the control group administered distilled water alone, Low-dose represents the low-dose group administered 360 mg CG/kg body weight (corresponding to 0.31 mg sodium cyanide/kg body weight), and High-dose represents the high-dose group administered 2840 mg CG/kg body weight (corresponding to 2.44 mg sodium cyanide/kg body weight). Bars represent means ± standard deviation (3 rats/group). Asterisks indicate significant differences from control values (**p < 0.01).

The bar represents the change of thiocyanate level in the hemolyzed blood from 0 h to 8 h after administration of CG

The bar represents the change of thiocyanate level in the hemolyzed blood from 0 h to 24 h after administration of CG

Fig. 4.

Thiocyanate content in the brains of adenine-induced CRD rats.

Control represents the control group administered distilled water only, Low-dose represents the low-dose group administered 360 mg CG/kg body weight (corresponding to 0.31 mg sodium cyanide/kg body weight), and High-dose represents the high-dose group administered 2840 mg CG/kg body weight (corresponding to 2.44 mg sodium cyanide/kg body weight). Bars represent means ± standard deviation (3 rats/group). Asterisks indicate significant differences from control values (*p < 0.05).

The bar represents brain thiocyanate levels after administration of CG

Fig. 5.

Macroscopy of the normal kidney (left), and Adenine represents the adenine-induced crystal nephropathy observed in CRD rats (right).

Fig. 6.

Histopathology of the cerebrum cortex and cerebrum caudal nucleus of CRD rats after administration of Sugihiratake extract.

Control represents the control group administered distilled water alone, and High-dose represents the high-dose group administered 2840 mg CG/kg body weight (corresponding to 2.44 mg sodium cyanide/kg body weight. Microscopically no treatment related abnormalities were observed in neurons and glial cells. x100. Hematoxylin-eosin staining.

Fig. 7.

Histopathological images of the kidneys of CRD rats.

A-D, Histopathological changes in the kideny. A and C, Normal structure obtained from a normal adult rat kidney. B and D, Adenine-induced chronic renal damage. The normal structure was destroyed, and brown to black dots, adenine crystals, were diffusely distributed throughout the kidney at low magnification (x10). C and D, Higher magnification (x100) of A and B. C. Normal structure. D, Various sizes of crystals are accumulated in the dilated lumen of renal tubules or deposited in the tubules. Inflammatory cells infiltrate the interstitium. Hematoxylin-eosin staining.

Tables

Table 1.The body weight changes

IndividualNo.	Animal*No.		Quarantine and acclimation periods			Adenine-component feed feeding period
						---- 0.75% ---		-------- 0.5% --------
		Day	1	5	9	1	8	15	22	28
1	1302		298	334	355	362	281	270	270	259
2	exc.		308	355	381	384	267	-	-	-
3	1203		295	344	366	368	268	282	285	282
4	1103		288	317	332	334	257	220	219	220
5	1301		304	337	368	378	270	282	285	256
6	1303		297	343	366	375	321	303	298	282
7	1101		315	354	385	388	293	258	260	262
8	1201		306	348	368	378	301	279	279	261
9	1102		303	340	361	362	275	271	270	273
10	1202		305	345	363	369	266	268	264	254
Mean			302	342	365	370	280	270	270	261
S.E.			2	3	5	5	6	8	8	6

Unit: g.

No.2: died on day 8 of adenine-component feed feeding period (266 g body weight).

^* Identification numbers after allocation to treatment groups (exc.: excluded from this study).

Quarantine period: 5 days.

Acclimation period: 9 days (with quarantine period).

Table 2.Pathological findings in rats with adenine-induced renal failure after oral administration of Sugihiratake extract

	Test substance	Control				Sugihiratake extract
						360 mg/kg				2840 mg/kg
	Animal No.	1101	1102	1103		1201	1202	1203		1301	1302	1303
Heart
Myocardium
Focal calcification		0	0	1		1	0	0		1	1	0
Coronary artery
Cellular infiltration, mononuclear cell		1	0	1		1	0	0		1	1	0
Calcification		0	0	3		3	0	0		3	2	1
Arcus aorta
Tunica media, calcification		2	0	3		3	1	0		3	2	1
Tunica intima, cellular infiltration, neutrophil / mononuclear cell		1	0	0		1	0	0		0	0	0
Lung
Alveolar septa, calcification		0	0	3		0	0	0		2	0	0
Bronchial smooth muscle, calcification		0	0	2		0	0	0		1	0	0
Kidney
Artery
Calcification		1	0	2		2	1	0		2	2	1
Renal tubule
2,8-dihydroxyadenine (DHA) crystals		3	3	3		3	3	3		3	3	3
Tubular dilatation		3	3	3		3	3	3		3	3	3
Tubular basophilic change		3	3	3		3	3	3		3	3	3
Cellular infiltration, neutrophil		2	2	2		2	2	2		2	2	2
Giant cells		1	1	1		1	1	1		1	1	1
Interstitium
Cellular infiltration, neutrophil / mononuclear cell		1	1	1		1	1	1		1	1	1
Fibrosis		2	2	2		2	2	2		2	2	2

0: No change, 1: Slight, 2: Moderate, 3: Marked.

No significant changes were detected: Spleen, Liver.

Table 3.Pathological findings in rats with adenine-induced renal failure after oral administration of Sugihiratake extract

	Test substance	Control				Sugihiratake extract
						360 mg/kg				2840 mg/kg
Findings	Animal No.	1101	1102	1103		1201	1202	1203		1301	1302	1303
Kidney
Enlargement, pale yellow		+	+	+		+	+	+		+	+	+
Thymus
Small size		+	+	+		+	+	+		+	+	+
Arcus aorte
Grayish white		-	-	+		-	-	-		-	-	-
Coronary artery
Grayish white		-	-	+		-	-	-		-	-	-
Glandular stomach
Grayish white		-	-	+		-	-	-		+	-	-
Colon
Grayish white		-	-	+		-	-	-		+	-	-

-: No change, +: Change.

References

• 1.  Gejyo F, Homma N, Higuchi N, et al. A novel type of encephalopathy associated with mushroom Sugihiratake ingestion in patients with chronic kidney diseases. Kidney Int. 2005; 68(1): 188–192.
• 2.  Suzuki T, Igarashi K, Dohra H, et al. A new omics data resource of Pleurocybella porrigens for gene discovery. PLoS One. 2013; 8(7): e69681.
• 3.  Suzuki T, Amano Y, Fujita M, et al. Purification, characterization, and cDNA cloning of a lectin from the mushroom Pleurocybella porrigens. Biosci Biotechnol Biochem. 2009; 73(3): 702–709.
• 4.  Kawaguchi T, Suzuki T, Kobayashi Y, et al. Unusual amino acid derivatives from the mushroom Pleurocybella porrigens. Tetrahedron. 2010; 66(2): 504–507.
• 5.  Takano F, Yamaguchi M, Shoda S, Fu ZD, Ohta T. Toxicological studies on hot water extracts of Pleurocybella porrigens (Pers.:Fr.) in mice. Natural Medicines. 2005; 59(4): 151–156.
• 6.  Smith ADM, Duckett S, Waters AH. Neuropathological changes in chronic cyanide intoxication. Nature. 1963; 200: 179–181.
• 7.  Smith ADM. Cyanide encephalopathy in man? Lancet. 1964; 2(7361): 668–670.
• 8.  Funata N, Song SY, Okeda R, Funata M, Higashino F. A study of experimental cyanide encephalopathy in the acute phase—physiological and neuropathological correlation. Acta Neuropathol (Berl). 1984; 64(2): 99–107.
• 9.  Rachinger J, Fellner FA, Stieglbauer K, Trenkler J. MR changes after acute cyanide intoxication. AJNR Am J Neuroradiol. 2002; 23(8): 1398–1401.
• 10.  Yen D, Tsai J, Wang LM, et al. The clinical experience of acute cyanide poisoning. Am J Emerg Med. 1995; 13(5): 524–528.
• 11.  Wilson J. Cyanide in human disease: a review of clinical and laboratory evidence. Fundam Appl Toxicol. 1983; 3(5): 397–399.
• 12.  Akiyama H, Toida T, Sakai S, et al. Determination of cyanide and thiocyanate in Sugihiratake mushroom using HPLC method with fluorometric detection. J Health Sci. 2006; 52(1): 73–77.
• 13.  Nagano N, Miyata S, Abe M, et al. Effect of manipulating serum phosphorus with phosphate binder on circulating PTH and FGF23 in renal failure rats. Kidney Int. 2006; 69(3): 531–537.
• 14.  Obara K, Okawa S, Kobayashi M, Takahashi S, Watanabe S, Toyoshima I. [A case of encephalitis-type encephalopathy related to Pleurocybella porrigens (Sugihiratake)]. Rinsho Shinkeigaku. 2005; 45(3): 253–256. Japanese.
• 15.  Kuwabara T, Arai A, Honma N, Nishizawa M. [Acute encephalopathy among patients with renal dysfunction after ingestion of “sugihiratake”, angel’s wing mushroom—study on the incipient cases in the northern area of Niigata Prefecture]. Rinsho Shinkeigaku. 2005; 45(3): 239–245. Japanese.
• 16.  Kurokawa K, Sato H, Nakajima K, Kawanami T, Kato T. [Clinical, neuroimaging and electroencephalographic findings of encephalopathy occuring after the ingestion of “sugihiratake” (Pleurocybella porrigens), an autumn mashroom: a report of two cases]. Rinsho Shinkeigaku. 2005; 45(2): 111–116. Japanese
• 17.  Kato T, Kawanami T, Shimizu H, et al. [An outbreak of encephalopathy after eating autumn mushroom (Sugihiratake; Pleurocybella porrigens) in patients with renal failure: a clinical analysis of ten cases in Yamagata, Japan]. No To Shinkei. 2004; 56(12): 999–1007. Japanese.
• 18.  Nomoto T, Seta T, Nomura K, et al. A case of reversible encephalopathy accompanied by demyelination occurring after ingestion of Sugihiratake mushrooms. J Nippon Med Sch. 2007; 74(3): 261–264.
• 19.  Eyjólfsson R. Recent advances in the chemistry of cyanogenic glycosides. Fortscher Chem Org Naturst. 1970; 28: 74–108.
• 20.  Singer R. The Agaricales in modern taxonomy. 3rd ed. J. Cramer Vaduz 1975; 912.
• 21.  Spencer PS. Food toxins, ampa receptors, and motor neuron diseases. Drug Metab Rev. 1999; 31(3): 561–587.
• 22.  Knowles CJ. Microorganisms and cyanide. Bacteriol Rev. 1976; 40(3): 652–680.
• 23.  Hasuike Y, Nakanishi T, Moriguchi R, et al. Accumulation of cyanide and thiocyanate in haemodialysis patients. Nephrol Dial Transplant. 2004; 19(6): 1474–1479.
• 24.  Pettigrew AR, Fell GS. Simplified colorimetric determination of thiocyanate in biological fluids, and its application to investigation of the toxic amblyopias. Clin Chem. 1972; 18(9): 996–1000.
• 25.  Andersen SL, Nøhr SB, Wu CS, Olsen J, Pedersen KM, Laurberg P. Thyroglobulin in smoking mothers and their newborns at delivery suggests autoregulation of placental iodide transport overcoming thiocyanate inhibition. Eur J Endocrinol. 2013; 168(5): 723–731.
• 26.  Wang H, Sekine M, Yokokawa H, et al. The relationship between new stroke onset and serum thiocyanate as an indicator to cigarette smoking. J Epidemiol. 2001; 11(5): 233–237.
• 27.  Arai A, Silberg J, Kessler M, Lynch G. Effect of thiocyanate on AMPA receptor mediated responses in excised patches and hippocampal slices. Neuroscience. 1995; 66(4): 815–827.
• 28.  Nielsen EO, Johansen TH, Wätjen F, Drejer J. Characterization of the binding of [3H]NS 257, a novel competitive AMPA receptor antagonist, to rat brain membranes and brain sections. J Neurochem. 1995; 65(3): 1264–1273.
• 29.  Hawkinson JE, Espitia SA. Effects of thiocyanate and AMPA receptor ligands on (S)-5-fluorowillardiine, (S)-AMPA and (R,S)-AMPA binding. Eur J Pharmacol. 1997; 329: 213-221.
• 30.  Ross SM, Roy DN, Spencer PS. β-N-oxalylamino-L-alanine action on glutamate receptors. J Neurochem. 1989; 53(3): 710–715.
• 31.  Murphy DE, Snowhill EW, Williams M. Characterization of quisqualate recognition sites in rat brain tissue using DL-[3H]alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) and a filtration assay. Neurochem Res. 1987; 12(9): 775–781.
• 32.  Nielsen EO, Cha JH, Honoré T, Penney JB, Young AB. Thiocyanate stabilizes AMPA binding to the quisqualate receptor. Eur J Pharmacol. 1988; 157(2-3): 197–203.
• 33.  Baudry M, Monaghan D, Cotman C, Altar CA. Regional distribution of alpha-[3H]amino-3-hydroxy-5-methylisoxazole-4-propionic acid binding sites in rat brain: effect of chemical modification of SH- groups in tissue sections. J Neurochem. 1990; 54: 1682-1688.
• 34.  Hall RA, Massicotte G, Kessler M, Baudry M, Lynch G. Thiocyanate equally increases affinity for two DL-α-amino-3-hydroxy-5-methylisoxazolepropionic acid (AMPA) receptor states. Mol Pharmacol. 1993; 43(3): 459–464.
• 35.  Shahi K, Baudry M. Increasing binding affinity of agonists to glutamate receptors increases synaptic responses at glutamatergic synapses. Proc Natl Acad Sci USA. 1992; 89(15): 6881–6885.
• 36.  Cha JH, Makowiec RL, Penney JB, Young AB. Multiple states of rat brain (RS)-α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptors as revealed by quantitative autoradiography. Mol Phrmacol. 1992; 41(5): 832–838.
• 37.  Nessim SJ, Richardson RM. Dialysis for thiocyanate intoxication: a case report and review of the literature. ASAIO J. 2006; 52(4): 479–481.