Discovery of bioactive compounds in the Chinese herbal formula NaoShuanTong Capsule (NSTC) against hemorheological disorders

Figures & data

Table 1. Adjusted formulations of NSTC samples based on uniform design.

NSTC samples	Herb weight ratio (%)
	PT	RPR	RC	RG	RR
NSTC1	47.5	0	9.5	15	28
NSTC2	38.25	6	23.75	7.5	24.5
NSTC3	29	12	38	0	21
NSTC4	42.25	18	4.75	17.5	17.5
NSTC5	33	24	19	10	14
NSTC6	23.75	30	33.25	2.5	10.5
NSTC7	37	36	0	20	7
NSTC8	27.75	42	14.25	12.5	3.5
NSTC9	18.5	48	28.5	5	0

Table 2. Peak areas of 10 compounds in the HPLC fingerprint of NSTC samples.

Samples	P1	P2	P3	P4	P5	P6	P7	P8	P9	P10
NSTC1	17.861	17.635	1.365	1.152	0.800	0.000	0.000	4.051	11.475	11.655
NSTC2	11.283	10.930	20.255	2.543	11.201	1.339	1.509	1.793	6.581	6.977
NSTC3	7.343	6.708	28.340	3.854	17.103	1.531	2.183	0.000	4.583	4.884
NSTC4	10.419	11.190	45.942	6.598	28.759	3.120	5.726	3.677	5.164	4.538
NSTC5	6.646	6.268	47.201	6.805	29.572	3.488	5.832	1.080	3.232	2.849
NSTC6	4.402	4.025	51.278	7.124	30.772	4.077	5.734	0.557	2.160	1.777
NSTC7	6.346	6.621	63.321	8.935	38.630	6.179	8.570	3.048	1.996	1.114
NSTC8	5.100	4.838	66.527	9.652	41.543	6.544	8.748	1.944	1.156	0.000
NSTC9	3.501	3.589	77.076	9.834	46.692	6.345	10.270	1.509	0.000	0.000

Note: The compounds were as follows: P1, typhaneoside; P2, isorhamnetin-3-O-neohesperidoside; P3, gallic acid; P4, oxypaeoniflorin; P5, paeoniflorin; P6, galloyl-paeoniflorin; P7, benzoyl-oxypaeoniflorin; P8, gastrodin; P9, ajugasterone-C; P10, β-ecdysterone. P1, P2, P5, and P10 were identified by authentic standards, the other compounds were characterized by MS data and literature.

Table 3. Effects of nine NSTC samples on haemorheological parameters.

Group	Concentration (mg/kg)	WBV 5/s	WBV 30/s	WBV 50/s	WBV 150/s	WBV 200/s	EAI	EEI	EDI	PT (s)	APTT (s)	PV 120/s	MPAR
Control	NS	9.05 ± 2.01	6.18 ± 0.46	5.58 ± 0.37	4.78 ± 0.30	4.49 ± 0.17	1.72 ± 0.16	4.01 ± 0.67	1.03 ± 0.05	8.74 ± 0.28	14.04 ± 1.03	0.99 ± 0.02	33.33 ± 4.20
Model	NS	11.48 ± 1.97^#	7.52 ± 0.80^##	6.75 ± 0.66^##	5.32 ± 0.38^#	4.92 ± 0.34^#	2.12 ± 0.18^##	4.74 ± 0.29^##	0.93 ± 0.04^##	7.98 ± 0.25^##	12.29 ± 0.93^##	1.17 ± 0.02^##	40.33 ± 3.24^##
ASP	100	8.41 ± 2.76*	6.28 ± 0.93*	5.73 ± 0.72*	4.96 ± 0.38	4.86 ± 0.39	1.58 ± 0.26**	3.53 ± 0.82	0.94 ± 0.02	8.13 ± 0.30	13.01 ± 0.70	1.16 ± 0.02	31.47 ± 6.19*
NSTC1	1600	8.98 ± 1.66*	6.60 ± 0.65*	6.31 ± 0.86	5.35 ± 0.29	5.14 ± 0.35	1.62 ± 0.20**	3.57 ± 0.46*	0.95 ± 0.04	8.10 ± 0.29	12.81 ± 1.09	1.16 ± 0.01	36.41 ± 3.21
NSTC2	1600	8.28 ± 1.66**	6.50 ± 1.00*	5.80 ± 0.45**	5.36 ± 0.37	5.10 ± 0.28	1.66 ± 0.26**	3.61 ± 0.71*	0.96 ± 0.05	7.99 ± 0.31	13.15 ± 1.56	1.15 ± 0.04	36.31 ± 1.77*
NSTC3	1600	9.06 ± 1.75*	6.68 ± 0.72	6.34 ± 0.44	5.37 ± 0.22	4.86 ± 0.41	1.56 ± 0.12**	3.37 ± 0.26*	1.03 ± 0.10*	7.99 ± 0.31	12.83 ± 1.20	1.17 ± 0.01	38.56 ± 6.12
NSTC4	1600	8.03 ± 2.40**	6.36 ± 0.51**	5.74 ± 0.46**	5.01 ± 0.36	5.11 ± 0.47	1.76 ± 0.21**	3.74 ± 0.40**	0.96 ± 0.04	8.04 ± 0.40	13.50 ± 1.98	1.17 ± 0.02	40.86 ± 5.63
NSTC5	1600	7.70 ± 1.00**	6.23 ± 0.53**	5.64 ± 0.58**	4.92 ± 0.33*	5.13 ± 0.33	1.85 ± 0.27**	4.12 ± 0.67**	0.95 ± 0.04	7.93 ± 0.27	13.29 ± 1.86	1.14 ± 0.02*	34.54 ± 4.55*
NSTC6	1600	10.37 ± 2.62	7.15 ± 0.81	6.51 ± 1.00	5.45 ± 0.64	4.92 ± 0.34	1.85 ± 0.14	4.03 ± 0.45	0.99 ± 0.04	8.15 ± 0.50	13.76 ± 1.15*	1.14 ± 0.03*	36.42 ± 2.67*
NSTC7	1600	8.87 ± 2.02*	6.59 ± 0.63*	6.17 ± 0.68	5.12 ± 0.39	5.15 ± 0.25	1.72 ± 0.42*	3.91 ± 0.82**	0.95 ± 0.03	7.90 ± 0.35	13.38 ± 1.71	1.17 ± 0.02	40.21 ± 3.60
NSTC8	1600	9.89 ± 1.86	6.98 ± 0.72	6.50 ± 0.65	5.28 ± 0.37	5.27 ± 0.35	1.92 ± 0.28**	4.23 ± 0.61*	0.92 ± 0.04	8.10 ± 0.37	13.80 ± 0.80**	1.16 ± 0.01	36.79 ± 5.56
NSTC9	1600	9.89 ± 1.34	6.89 ± 0.34	6.40 ± 0.63	5.27 ± 0.43	4.99 ± 0.40	1.75 ± 0.25*	3.95 ± 0.67	0.99 ± 0.06*	8.11 ± 0.45	13.80 ± 1.48*	1.17 ± 0.03	40.98 ± 6.45

Note: All data are mean values ± SD, n = 10. WBV and PV unit: mPa.s; APTT and PT unit: second (s).

^# P < 0.05; ^##P < 0.01, vs. control group.

^* P < 0.05; ^**P < 0.01, vs. model group.

Table 4. Total variance explained by components X1–X4.

Component	Variance %	Accumulated variance %
X1	46.716	46.716
X2	17.944	64.660
X3	16.443	81.103
X4	15.018	96.122

Table 5. Rotated component matrix.

Parameters	X1	X2	X3	X4
PT	0.136	−0.19	0.951	−0.106
APTT	−0.33	0.072	0.898	0.244
WBV 5/s	0.969	−0.111	−0.076	0.195
WBV 30/s	0.948	−0.097	−0.008	0.274
WBV 50/s	0.931	−0.115	−0.093	0.188
WBV 150/s	0.638	−0.268	0.376	0.522
WBV 200/s	−0.183	0.97	−0.095	−0.071
PV	0.496	−0.08	0.221	0.788
EAI	0.958	−0.095	0.065	0.253
EDI	−0.085	0.979	−0.042	−0.148
EEI	0.928	−0.149	−0.101	0.238
MPAR	0.548	−0.26	−0.184	0.736

Table 6. Scores of components.

NSTC samples	X1	X2	X3	X4
NSTC1	2.010	−0.304	19.197	4.239
NSTC2	2.268	−0.330	18.894	4.188
NSTC3	1.729	−0.166	19.569	4.255
NSTC4	2.210	−0.310	19.258	4.351
NSTC5	2.186	−0.343	19.884	4.473
NSTC6	1.349	−0.133	19.361	4.140
NSTC7	1.878	−0.290	19.882	4.401
NSTC8	1.361	−0.239	19.939	4.265
NSTC9	1.262	−0.144	20.567	4.453

Table 7. Chi-square test results.

Set of canonical variables	Canonical correlation coefficient	Degrees of freedom	χ^2 values	P-values of likelihood ratio chi-square
1	0.996	40	94.7869	2.42 × 10^-6
2	0.9616	27	49.0535	0.0058
3	0.9175	16	24.6868	0.0755
4	0.7198	7	7.1380	0.4147

Table 8. CCA model between compounds and effects.

Set of canonical variables	CCA model
1	$\begin{array}{l}{V}_1=0.266y_1+0.616y_2+0.326y_7\\ W_1=7.215X_1\end{array}$
2	$V_2=0.937y_2+0.249y_3+2.673y_7+1.444y_9{W}_2=28.142X_4$
3	$\begin{array}{l}{V}_3=0.617y_2+0.425y_5+0.951y_8+1.674y_{10}-1.917y_1-0.701y_6-0.706y_7\\ W_3=22.332X_2-4.578X_3\end{array}$

Table 9. Corelation between parameters and components.

Effect component	Correlated components	Correlation coefficient
Erythrocyte aggregation	Typhaneoside^a	0.266
	Isorhamnetin-3-O-neohesperidoside^b	0.616
	Benzoyl-oxypaeoniflorin^c	0.326
Erythrocyte deformability	Isorhamnetin-3-O-neohesperidoside^b	0.617
	Paeoniflorin	0.425
	Gastrodin	0.951
	β-Ecdysterone	1.674
Coagulation function	Typhaneoside^a	1.917
	Galloyl-paeoniflorin	0.701
	Benzoyl-oxypaeoniflorin^c	0.706
PV and MPAR	Isorhamnetin-3-O-neohesperidoside^b	0.937
	Gallic acid	0.249
	Benzoyl-oxypaeoniflorin^c	2.673
	Ajugasterone-C	1.444

Note: Superscript letters indicate compounds that were significantly correlated with more than one effect parameter.

Figure 1. HPLC fingerprints of NSTC samples.

Figure 2. Cluster analysis of NSTC samples.