Increased cardiac distribution of mono-PEGylated Radix Ophiopogonis polysaccharide in both myocardial infarction and ischemia/reperfusion rats

Figures & data

Figure 1 Representative chromatograms.

Notes: Representative chromatograms for the determination of ^FMP_20k-R (A) and ^FMP_40k-R (B) in the heart tissue by HPGPC. 1= blank heart tissue; 2= blank heart tissue spiked with standard ^FMP_20k-R solution; 3= heart tissue sample collected at 3 hours after intravenous administration of ^FMP_20k-R; 4= blank heart tissue spiked with standard ^FMP_40k-R solution; 5= heart tissue sample collected at 3 hours after intravenous administration of ^FMP_40k-R.

Abbreviations: ^FMP_20k-R, 20-kDa PEG mono-modified ROP; ^FMP_40k-R, 40-kDa PEG mono-modified ROP; HPGPC, high-performance gel permeation chromatography; PEG, polyethylene glycol; ROP, Radix Ophiopogonis polysaccharide.

Table 1 Linearity and sensitivity of detection for ^FMP_20k-R and ^FMP_40k-R in rat myocardia by HPGPC (n=7)

Samples	Standard curves	Linear ranges (μg/mL)	Correlation coefficients	DL (μg/mL)	QL (μg/mL)
^FMP20k-R	H=54.867C-0.051	1.42–91.09	0.9997	0.28	0.94
^FMP40k-R	H=33.212C-0.083	5.79–370.91	0.9994	0.44	1.45

Abbreviations: DL, detection limit; ^FMP_20k-R, 20-kDa PEG mono-modified ROP; ^FMP_40k-R, 40-kDa PEG mono-modified ROP; HPGPC, high-performance gel permeation chromatography; PEG, polyethylene glycol; QL, quantification limit; ROP, Radix Ophiopogonis polysaccharide.

Table 2 Recovery, accuracy, and precision for the assay of ^FMP_20k-R and ^FMP_40k-R in rat myocardia by HPGPC (n=5)

Samples	Added concentration (μg/mL)	Recovery (%)	Accuracy (%)	Precision RSD (%)
				Within-batch	Inter-batch
^FMP20k-R	2.85	55.46±2.13	97.39±6.87	10.61	8.87
	11.39	59.49±3.68	99.94±2.42	2.63	4.76
	45.55	65.91±1.79	100.78±1.23	1.24	2.04
^FMP40k-R	11.59	48.02±5.70	97.72±9.03	9.20	15.65
	46.36	54.16±5.78	94.08±9.46	5.93	11.75
	185.45	59.74±4.24	97.85±6.86	6.69	6.09

Abbreviations: ^FMP_20k-R, 20-kDa PEG mono-modified ROP; ^FMP_40k-R, 40-kDa PEG mono-modified ROP; HPGPC, high-performance gel permeation chromatography; PEG, polyethylene glycol; ROP, Radix Ophiopogonis polysaccharide; RSD, relative standard deviation.

Figure 2 Representative TTC staining photographs.

Notes: Representative TTC staining photographs of heart transverse slices from a rat receiving IR surgery with reperfusion for 24 hours (A, infarct size =43.4%) and slices from a rat receiving MI surgery with a prolonged period of ischemia for 4 days (B, infarct size =19.2%).

Abbreviations: IR, ischemia/reperfusion; MI, myocardial infarction; TTC, triphenyltetrazolium chloride.

Table 3 Concentrations of ^FMP_20k-R in myocardium and plasma after a single intravenous dose of 4 μmol/kg in rats (n=4–5)

Biosample	Group	Concentration (μmol/L)
		3 hours	12 hours	24 hours
Myocardium	Normal	0.535±0.051	0.267±0.021	0.159±0.023
	IR	1.222±0.149**	0.372±0.041**	0.227±0.020**
	MI	0.876±0.134**^,Table Footnote##	0.384±0.086**	0.332±0.012**^,Table Footnote##
Plasma	Normal	18.11±1.25	4.54±0.94	1.51±0.23
	IR	20.42±5.23	4.22±0.53	1.63±0.14
	MI	15.14±0.74*	3.08±0.47*^,#	1.60±0.07

Notes:

** P<0.01

* P<0.05, compared with the normal group.

## P<0.01

# P<0.05, compared with the IR group.

Abbreviations: ^FMP_20k-R, 20-kDa polyethylene glycol mono-modified Radix Ophiopogonis polysaccharide; IR, ischemia/reperfusion; MI, myocardial infarction.

Table 4 Concentrations of ^FMP_40k-R in myocardium and plasma after a single intravenous dose of 4 μmol/kg in rats (n=4–5)

Biosample	Group	Concentration (μmol/L)
		3 hours	12 hours	24 hours
Myocardium	Normal	0.944±0.105	0.789±0.078	0.407±0.064
	IR	1.739±0.233**	1.159±0.126**	0.785±0.096**
	MI	1.367±0.166**^,Table Footnote#	1.137±0.188**	0.894±0.094**
Plasma	Normal	50.63±4.53	25.66±1.70	17.68±1.33
	IR	57.34±2.14	21.95±3.89	15.05±1.70
	MI	50.47±1.62##	18.78±2.31**	15.50±2.61

Notes:

** P<0.01, compared with the normal group.

## P<0.01

# P<0.05, compared with the IR group.

Abbreviations: ^FMP_40k-R, 40-kDa polyethylene glycol mono-modified Radix Ophiopogonis polysaccharide; IR, ischemia/reperfusion; MI, myocardial infarction.

Table 5 Cardiac distribution tendency of ^FMP_20k-R in normal, IR, and MI rats and cardiac targeting efficacy of ^FMP_20k-R in IR and MI rats at predetermined times postdose (n=4–5)

Time	Distribution tendency (%)			Targeting efficacy
	Normal	IR	MI	IR	MI
3 h	2.96±0.22	6.22±1.28**	5.79±0.80**	2.10±0.43	1.96±0.27
12 h	6.01±0.87	8.82±0.26**	12.54±2.70*	1.47±0.04$$	2.11±0.45
24 h	10.65±1.31	14.06±2.12**	20.74±1.22**^,##	1.32±0.20$$	1.95±0.11&&

Notes:

** P<0.01

* P<0.05, compared with the normal group;

## P<0.01, compared with the IR group;

$$ P<0.01, compared with the targeting efficacy at 3 hours postdose;

&& P<0.01, compared with the IR group at 24 hours.

Abbreviations: ^FMP_20k-R, 20-kDa polyethylene glycol mono-modified Radix Ophiopogonis polysaccharide; IR, ischemia/reperfusion; MI, myocardial infarction.

Table 6 Cardiac distribution tendency of ^FMP_40k-R in normal, IR, and MI rats and cardiac targeting efficacy of ^FMP_40k-R in IR and MI rats at predetermined times postdose (n=4–5)

Time	Distribution tendency (%)			Targeting efficacy
	Normal	IR	MI	IR	MI
3 h	1.88±0.31	3.04±0.44**	2.71±0.27*	1.61±0.23	1.44±0.14
12 h	3.07±0.22	5.35±0.62**	6.08±0.94*	1.74±0.20	1.98±0.31
24 h	2.30±0.34	5.22±0.16**	5.91±1.29**	.26±0.07^$$,##	2.57±0.56$$

Notes:

** P<0.01

* P<0.05, compared with the normal group;

$$ P<0.01, compared with the targeting efficacy at 3 hours;

## P<0.01, compared with the targeting efficacy at 12 hours.

Abbreviations: ^FMP_40k-R, 40-kDa polyethylene glycol mono-modified Radix Ophiopogonis polysaccharide; IR, ischemia/reperfusion; MI, myocardial infarction.

Figure 3 Cardiac distribution tendency.

Figure 4 Cardiac targeting efficacy.

Note: Cardiac targeting efficacy of ^FMP_20k-R (A) and ^FMP_40k-R (B) at predetermined times postdose in the rat models of MI and IR.

Abbreviations: ^FMP_20k-R, 20-kDa PEG mono-modified ROP; ^FMP_40k-R, 40-kDa PEG mono-modified ROP; IR, ischemia/reperfusion; MI, myocardial infarction; PEG, polyethylene glycol; ROP, Radix Ophiopogonis polysaccharide.

Note: Cardiac distribution tendency of ^FMP_20k-R (A) and ^FMP_40k-R (B) versus time profiles in normal, IR, and MI rats.

Abbreviations: ^FMP_20k-R, 20-kDa PEG mono-modified ROP; ^FMP_40k-R, 40-kDa PEG mono-modified ROP; IR, ischemia/reperfusion; MI, myocardial infarction; PEG, polyethylene glycol; ROP, Radix Ophiopogonis polysaccharide.

Figure 4 Cardiac targeting efficacy.

Note: Cardiac targeting efficacy of ^FMP_20k-R (A) and ^FMP_40k-R (B) at predetermined times postdose in the rat models of MI and IR.

Abbreviations: ^FMP_20k-R, 20-kDa PEG mono-modified ROP; ^FMP_40k-R, 40-kDa PEG mono-modified ROP; IR, ischemia/reperfusion; MI, myocardial infarction; PEG, polyethylene glycol; ROP, Radix Ophiopogonis polysaccharide.