Chronic toxicity of both raw and processed Polygoni Multiflori Radix on rats

doi:10.5246/jcps.2016.01.006

Abstract

Abstract:

Recently, adverse effects of Polygoni Multiflori Radix (PMR) and Polygoni Multiflori Radix Praeparata(PMRP)have attracted intensive attention worldwide. These adverse effects most occurred in caseswith high dose of prolonged medication course.Liver is usually the target organ of these adverse effects. In the present research,we performedin vivo chronic toxicity study and aimed to evaluate relationships between major constituents of water extractions and total anthraquinone of PMR and PMRP and chronic toxicity.SD rats of both sexes were given water extractions as well as total anthraquinone of PMR and PMRP for 12 weeks.We evaluated basic biochemical indexes, conducted microscopic observations of main organs and assessed early indicators of liver and renal fibrosis. Simvastatin, with hypertriglyceridemia and hypercholesterolemia as its main therapeutic areas, was investigated in our study. Component-toxicity relationships were also discussed.Five male rats died in our study, while all female rats survived, suggesting that some gender differences might be involved. Body weightwas significantly changed, and basic biochemical indexes were sporadically occurred during the research. Pathological examinations on liver and kidney showed slight alternations after 12 weeks without dose-dependent relationship. Increase in serum laminin (LN) was observed in almost all male rats, indicating that the risks of liver or kidney fibrosis still existed, especially for males, although no fibrosis was found in the pathological examination of liver and kidney.No major and severe adverse effects were observed after 12 weeks of administration of PMR and PMRP. Regular safety monitoring is still necessary during medication in order to prevent possible risks.

Key words: Chronic toxicity, Polygoni Multiflori Radix, Polygoni Multiflori Radix Praeparata, Total anthraquinone

CLC Number:

R284

Supporting:

Levels of serum biochemical indexes and HPCIII during the research.

Additional Table 1 ALT contents in female rats in this research (X±SD, n≥2) (U/L).

Groups	Week
Groups	3	4	5	6	7	8	9	10	11	12
A	17.96±10.27	37.33±7.767	40.33±6.658	38.33±9.018	38.00±1.414	44.50±0.7071	38.00±4.243	35.50±7.778	35.00±1.414	40.00±8.485
B	23.74±17.20	40.25±5.737	44.50±9.434	41.75±8.221	44.25±6.076	45.00±8.544	38.25±7.932	38.00±20.67	33.00±2.646	44.75±7.136
C	30.15±12.81	55.00±20.67	42.75±6.850	38.75±9.215	42.75±9.323	41.75±7.848	38.25±7.932	40.75±7.805	45.00±15.87	35.25±12.45
D	22.46±15.02	42.00±10.23	41.75±10.24	40.75±15.84	48.50±17.77	42.25±14.29	43.00±12.99	44.00±14.93	64.67±38.59	66.67±24.17
E	27.00±16.57	47.25±6.130	47.75±9.180	45.50±10.38	45.75±3.300^*	45.00±11.40	31.50±19.00	34.75±17.21	27.25±16.34	40.25±1.260
F	30.25±8.690	52.00±17.53	46.50±12.23	41.25±6.850	42.00±4.550	50.00±16.51	36.75±8.730	49.00±18.42	44.25±11.44	50.75±16.34
G	6.000±14.93	39.00±5.570	39.00±6.080	41.67±2.080	45.33±3.790	38.00±0^*	37.67±4.510	40.00±6.000	26.33±23.46	30.67±27.74
H	23.74±5.291	40.25±1.258	39.50±5.972	37.33±8.327	37.00±7.439	37.25±10.14	36.00±7.071	34.75±12.01	67.5±47.87	53.33±8.505
I	37.85±11.74	42.00±8.367	36.50±8.699	43.25±3.304	39.00±8.327	39.50±5.508	44.75±5.500	43.75±12.04	39.00±4.83	47.00±2.582
J	47.48±13.00^*	50.75±6.292	42.75±1.500	39.25±8.180	36.50±6.807	38.00±4.320	37.00±5.292	41.00±4.000	36.25±11.24	46.25±2.986

The ^* indicates a significant difference compared with control group. ^*p < 0.05.

Additional Table 2 ALT contents in male rats in this research (X±SD, n≥2) (U/L).

Groups	Week
Groups	3	4	5	6	7	8	9	10	11	12
A	35.29±15.16	41.00±3.464	45.33±3.512	49.67±1.528	48.33±4.619	44.67±5.686	46.67±8.145	50.67±10.02	48.67±3.786	57.00±4.583
B	38.50±10.05	45.00±6.733	39.67±7.767	39.33±4.163	48.50±3.536	50.50±4.950	53.00±15.56	23.00±22.63	53.50±16.26	49.50±3.536
C	31.44±4.858	41.50±8.813	46.00±3.742	51.33±5.033	43.50±7.937	42.25±3.304	37.25±13.79	44.00±5.000	44.67±9.018	59.33±4.163
D	42.99±5.691	45.25±7.890^*	52.00±8.524	45.75±2.630	42.50±5.447	48.50±8.813	46.25±3.304	51.75±5.909	50.25±11.03	54.25±17.35
E	29.50±6.790	51.00±4.360^*	41.67±4.163	48.00±2.000	49.00±4.243	48.00±1.414	48.50±3.536	53.00±24.04	47.50±7.778	50.00±14.69
F	54.54±32.35	56.67±5.859^*	51.00±5.568	41.67±15.03	62.00±18.43	39.00±11.21	47.00±9.530	49.00±13.25	47.00±10.48	67.00±19.64
G	32.75±7.663	48.75±4.031	48.00±9.381	45.67±9.074	44.50±8.813	38.75±9.535	37.50±10.66	69.67±25.72	42.67±3.215	52.67±4.509
H	41.70±8.479	51.50±6.758	48.75±7.500	53.50±13.30	47.75±8.057	42.50±12.69	41.25±10.75	69.00±15.00	43.00±36.59	62.00±11.36
I	44.27±14.12	40.50±9.037	48.75±3.304	47.25±9.323	44.00±4.967	44.25±6.131	30.67±18.34	42.00±12.25	51.00±4.830	57.25±7.805
J	34.00±17.95	43.25±9.912	49.50±14.80	45.25±14.03	44.00±17.07	42.00±9.626	42.50±7.681	95.75±103.6	46.50±13.82	53.00±9.899

The ^* indicates a significant difference compared with control group. ^*p < 0.05.

Additional Table 3 AST contents in female rats in this research (X±SD, n≥2) (U/L).

Groups	Week
Groups	3	4	5	6	7	8	9	10	11	12
A	53.67±20.99	88.67±14.01	111.7±23.50	89.67±7.095	67.00±2.828	78.00±5.657	81.00±12.73	86.00±11.31	77.00±9.899	116.5±47.38
B	66.08±49.69	113.0±30.17	167.3±72.73	129.3±32.68	138.8±56.93	103.3±21.67	109.0±28.37	124.0±15.53^*	97.00±14.14	139.5±39.50
C	102.7±34.79	115.3±39.14	169.3±65.64	83.75±13.40	112.0±62.46	90.25±22.62	109.0±28.37	121.8±29.34	161.3±100.8	137.5±74.30
D	69.08±9.071	123.3±14.24^*	139.0±10.86	148.5±51.70	132.0±46.45	117.8±19.00	122.0±34.21	125.0±20.95	154.5±71.42	147.7±23.18
E	75.09±16.69	120.0±19.61	166.3±35.94	148.3±30.05^*	130.5±21.11^*	123.8±17.93^*	127.3±17.35^*	114.5±22.07	97.25±23.41	137.3±32.83
F	53.46±33.55	114.0±43.63	146.8±36.81	110.3±20.65	110.3±22.68	92.25±43.01	110.5±56.77	122.3±26.06	93.00±13.89	159.3±25.91
G	59.27±8.439	140.7±50.52	146.0±59.63	100.0±10.00	91.67±5.859^*	65.67±44.81	113.0±12.17	127.0±52.09	107.5±37.48	176.5±24.75
H	85.30±27.78	97.00±21.34	125.8±28.44	107.3±23.84	117.5±23.46^*	114.5±23.17	86.25±47.89	136.5±51.20	100.3±36.69	177.0±54.06
I	103.3±23.46^*	88.75±19.43	128.5±27.45	101.3±16.40	109.8±19.26^*	91.00±11.78	85.25±11.93	104.3±27.66	86.25±12.69	157.3±24.96
J	94.31±41.13	109.0±13.59	149.5±50.05	97.25±14.64	93.00±6.218^*	80.00±7.874	85.25±7.805	103.5±31.30	103.8±34.27	164.8±18.19

The ^* indicates a significant difference compared with control group. ^*p < 0.05.

Additional Table 4 AST contents in male rats in this research (X±SD, n≥2) (U/L).

Groups	Week
Groups	3	4	5	6	7	8	9	10	11	12
A	114.1±100.7	116.7±34.99	183.7±86.41	108.7±10.12	110.7±7.095	108.0±12.12	117.7±28.10	156.0±35.59	140.3±32.15	177.0±13.89
B	126.8±47.51	153.0±51.74	202.3±41.79	130.0±2.828	116.5±16.26	119.0±2.828	134.5±55.86	76.00±60.81	82.00±82.02	174.5±9.192
C	71.49±18.86	121.0±27.65	174.8±22.56	127.0±41.35	118.3±14.86	107.5±6.758	120.5±24.35	121.0±24.04	122.3±4.163	206.7±62.68
D	101.5±40.08	123.0±10.23	287.5±40.14	136.5±19.47	120.3±27.10	113.0±12.36	109.5±18.36	143.8±29.34	104.5±56.44	173.5±27.21
E	83.50±10.27	125.0±15.10	129.08±13.42	116.0±12.29	113.5±16.26	126.5±23.33	145.5±0.7071	137.5±19.09	108.5±27.58	173.0±23.91
F	120.1±44.22	168.8±92.40	310.0±210.7	131.3±41.23	110.0±27.12	39.00±10.03	185.0±52.68	96.00±23.74	116±36.92	159.0±46.45
G	111.7±55.65	129.0±19.10	247.8±52.84	157.1±92.38	120.8±18.39	120.5±20.37	146.8±41.48	214.0±101.0	141.3±16.86	190.3±13.05
H	120.7±60.61	119.3±41.85	234.5±53.72	148.0±10.23^*	129.5±22.84	129.0±32.14	143.3±33.62	179.0±49.50	127.7±15.04	164.7±39.88
I	144.8±81.09	101.3±41.05	256.0±33.96	136.8±27.50	128.5±21.98	128.5±28.99	75.00±24.45	127.0±20.41	139.5±19.54	160.3±17.75
J	98.52±31.76	114.3±17.00	182.8±43.39	108.5±22.07	132.5±32.09	117.0±41.41	96.50±13.89	161.3±83.69	112.0±33.33	115.8±12.92^*

The ^* indicates a significant difference compared with control group. ^*p < 0.05.

Additional Table 5 ALP contents in female rats in this research (X±SD, n≥2) (U/L).

Groups	Week
Groups	3	4	5	6	7	8	9	10	11	12
A	161.2±57.00	150.7±55.77	166.3±70.03	138.0±52.12	156.0±38.18	163.0±59.40	156.0±79.20	109.5±27.58	131.5±48.79	167.0±53.74
B	409.7±273.4	323.5±277.6	296.3±231.6	206.0±167.2	246.0±175.0	228.0±148.7	95.67±28.29	164.0±103.5	121.7±62.5	226.8±142.0
C	103.8±25.18	121.5±26.16	107.3±20.66	99.25±26.63	93.75±16.58^*	102.0±14.99	95.67±28.29	86.50±13.03	86.25±11.53	134.3±8.083
D	138.0±42.17	191.5±92.18	95.50±51.03	190.0±124.3	260.5±279.9	180.0±110.5	189.3±125.7	113.7±72.86	113.0±65.85	154.3±76.25
E	130.9±41.82	168.0±23.17	149.5±46.02	146.3±50.03	135.3±45.22	144.8±39.88	126.3±56.55	103.0±41.99	97.00±33.71	112.8±72.31
F	129.5±22.94	157.0±42.83	124.8±30.57	102.8±26.42	116.8±27.71	121.0±33.44	134.3±36.49	96.25±25.88	100.8±8.770	103.3±17.06
G	125.2±29.57	200.0±90.16	178.7±112.2	174.3±76.51	147.3±66.40	143.3±62.16	136.7±18.88	114.7±34.65	130.0±11.31	286.5±160.5
H	112.4±9.718	106.5±6.608	70.50±8.888^*	100.5±14.55	124.3±76.57	122.0±34.28	89.50±9.110	93.75±18.52	83.25±11.67	135.3±23.29
I	146.5±58.29	170.5±84.25	101.3±39.48	155.3±58.05	129.5±44.20	163.3±64.20	115.5±46.96	120.0±51.00	112.3±44.25	132.3±86.42
J	240.4±31.64	272.3±38.32^*	195.3±36.64	170.5±34.07	157.3±30.82	185.5±41.81	161.0±43.60	146.0±31.02	152.8±56.62	133.8±39.12

The ^* indicates a significant difference compared with control group. ^*p < 0.05.

Additional Table 6 ALP contents in male rats in this research (X±SD, n≥2) (U/L).

Groups	Week
Groups	3	4	5	6	7	8	9	10	11	12
A	259.9±72.28	212.3±30.66	188.7±62.61	214.7±41.77	188.0±20.95	179.7±39.15	144.7±13.05	160.7±22.28	158.3±18.34	175.0±13.89
B	325.8±123.7	396.0±154.8	292.7±156.2	200.5±34.65	197.5±34.65	205.0±80.61	222.5±79.90	196.0±16.97	162.5±33.23	159.5±48.79
C	193.5±24.14	165.8±27.38	194.3±15.59	200.8±28.19	174.0±26.68	191.5±45.89	164.0±62.06	130.7±33.29	119.3±23.69	178.7±38.19
D	317.2±137.1	292.5±74.99	300.8±88.10	311.8±114.0	255.3±117.2	261.8±103.8	243.8±97.12	231.5±94.61	200.8±68.36	292.5±140.6
E	310.1±35.23	371.3±51.81^*	315.7±133.2	334.7±74.07	366.0±138.6	301.0±69.52	335.5±27.58^*	193.5±13.44	251.5±47.38^*	405.0±127.4
F	275.0±102.0	352.8±135.5	257.0±82.66	249.8±67.50	170.0±44.73	614.0±189.5	118.0±35.42	126.0±38.01	47.00±15.63	166.0±47.03
G	311.5±82.64	322.3±59.84^*	275.3±51.55	237.7±30.50	223.5±27.33	240.3±53.84	197.3±50.59	287.0±122.1	188.3±44.11	228.3±32.01
H	350.0±84.21	333.5±59.47^*	223.3±48.62	253.8±52.25	248.5±57.37	259.0±71.07	230.3±118.1	263.3±46.00^*	219.0±68.04	336.0±121.6
I	419.2±47.38^*	307.3±89.58	258.5±58.98	261.0±75.77	236.8±70.77	257.3±43.96	143.3±50.80	164.8±79.00	225.5±80.55	312.7±91.00
J	233.3±49.61	227.5±74.40	218.3±57.49	198.3±56.82	187.3±41.81	186.8±32.31	314.8±369.0	323.3±193.4	175.3±66.36	262.5±82.06

The ^* indicates a significant difference compared with control group. ^*p < 0.05.

Additional Table 7 TP contents in female rats in this research (X±SD, n≥2) (g/L).

Groups	Week
Groups	3	4	5	6	7	8	9	10	11	12
A	76.54±12.37	82.07±9.067	81.57±4.248	85.03±4.411	86.00±7.920	90.25±7.425	89.40±5.374	92.05±5.020	90.15±7.849	95.50±6.364
B	84.51±8.192	80.63±3.669	82.60±2.156	83.93±4.551	81.78±2.883	85.23±7.125	85.43±8.468	84.38±1.159^*	81.07±1.365	86.95±3.914
C	97.30±20.11	83.05±7.106	87.03±3.288	89.23±5.803	88.35±8.193	86.15±5.533	85.43±8.468	91.90±9.326	88.55±10.29	86.53±9.366
D	75.07±5.201	75.80±6.837	80.70±5.243	83.03±11.66	82.40±5.637	81.28±5.702	84.78±5.570	83.87±3.968	79.15±2.192	82.40±0.1414
E	83.29±6.825	83.33±1.907	86.65±4.655	84.03±3.661	86.20±1.426	84.58±2.401	84.70±4.320	73.83±29.55	86.98±1.832	83.45±1.103^*
F	80.40±15.01	81.73±3.837	81.93±5.527	82.18±2.748	85.70±3.212	85.38±3.788	81.48±9.703	85.18±10.22	88.67±3.253	90.78±5.151
G	86.69±7.949	79.30±7.641	81.67±6.216	87.83±5.486	87.27±1.504	82.03±1.457	83.23±1.305	89.40±2.651	88.20±1.414	83.35±0.2121
H	82.22±3.481	76.40±2.879	87.38±1.295^*	78.70±3.367	76.75±4.219	73.68±5.927^*	65.40±31.70	86.08±1.834	81.20±1.581	83.17±0.9866^*
I	77.35±5.286	79.58±6.896	92.15±3.724^*	85.05±1.115	87.48±7.350	84.18±2.948	91.30±6.082	93.83±5.658	85.55±4.324	86.45±5.157
J	77.05±8.164	80.05±9.259	83.70±6.526	83.38±2.861	82.90±5.356	86.05±9.234	87.75±9.386	92.08±9.280	83.60±6.815	83.85±7.238

The ^* indicates a significant difference compared with control group. ^*p < 0.05.

Additional Table 8 TP contents in male rats in this research (X±SD, n≥2) (g/L).

Groups	Week
Groups	3	4	5	6	7	8	9	10	11	12
A	73.09±1.611	71.78±6.134	73.80±2.364	76.23±5.676	75.63±3.496	74.77±8.237	76.13±3.453	79.90±5.620	75.47±4.219	78.87±3.361
B	76.44±10.27	72.33±6.987	76.60±2.651	77.05±0.07070	85.40±9.192	82.00±2.404	82.20±6.788	77.30±0	71.80±0.7071	78.10±1.556
C	93.04±11.88^*	68.68±3.899	73.75±4.516	77.23±3.313	76.00±4.531	77.10±3.536	76.60±6.092	78.10±4.667	73.60±1.054	76.40±3.822
D	81.01±9.630	71.35±0.7853	76.95±1.429	76.33±2.892	75.63±2.052	66.58±16.84	78.98±2.997	79.68±1.914	72.75±0.7326	72.50±10.26
E	82.68±7.264	73.73±3.782	81.53±4.450	79.90±2.425	75.00±2.970	76.55±2.333	75.85±3.465	81.55±0.4950	70.70±7.637	77.80±5.382
F	79.79±9.669	73.80±3.407	73.47±3.940	73.07±6.653	77.30±5.376	5.700±3.406	76.80±4.881	76.60±3.905	74.70±7.362	88.10±10.453
G	82.38±11.18	74.20±5.335	76.60±2.929	77.88±3.680	78.73±1.900	75.98±3.851	78.18±5.870	78.00±7.957	73.87±3.585	78.53±5.187
H	77.35±9.514	73.85±5.555	73.95±4.648	74.90±3.933	78.13±5.261	77.28±6.704	75.85±5.779	100.3±32.85	75.27±6.087	78.50±5.495
I	76.34±6.569	68.65±7.255	75.58±1.520	74.28±1.758	77.45±2.938	77.78±4.964	70.45±3.396	80.08±2.260	79.53±1.531	78.88±2.056
J	72.48±1.611	73.15±4.091	75.73±2.123	74.25±3.180	75.13±3.058	77.75±2.369	74.18±4.656	78.45±3.236	75.05±4.232	77.33±2.228

The ^* indicates a significant difference compared with control group. ^*p < 0.05.

Additional Table 9 BUN contents in female rats in this research (X±SD, n≥2) (mmol/L).

Groups	Week
Groups	3	4	5	6	7	8	9	10	11	12
A	6.911±0.0000	6.783±1.191	8.107±0.3855	6.960±0.3208	6.790±0.8627	7.695±0.03540	7.770±0.3111	7.665±1.761	5.320±0.5798	7.420±1.513
B	4.998±1.232^*	7.298±0.7991	6.345±0.9557^*	6.753±1.109	6.795±1.342	6.875±1.310	7.225±0.7527	7.420±0.9627	6.040±1.043	7.600±1.571
C	5.122±0.6171^*	7.338±1.150	6.878±0.6811^*	6.558±0.5382	6.948±0.3899	7.445±1.600	7.225±0.7527	7.350±0.5345	6.208±1.132	7.633±1.382
D	4.196±0.6373^*	6.810±0.5068	6.328±0.6108^*	7.710±1.441	6.463±0.6486	6.543±0.6469	7.170±1.174	7.597±0.6603	6.490±1.202	8.357±1.525
E	4.381±1.654^*	8.275±1.454	7.210±0.8189	9.263±0.5712^*	7.733±0.9514	7.805±0.4169	7.725±1.184	6.618±0.6319	6.358±0.9467	7.430±0.6328
F	5.122±1.232	7.903±1.053	6.508±0.8062^*	8.745±0.3478^*	7.465±0.6511	7.225±1.117	6.578±0.8689	6.120±0.6373	6.190±0.5717	7.515±1.297
G	4.525±1.995	5.263±0.8016	5.607±0.5886^*	6.650±0.4431	6.307±1.347	6.280±1.455	7.043±1.666	5.950±0.5751	6.580±0.2263	7.325±0.9546
H	6.541±0.8187	6.283±0.7509	8.168±0.8305	6.155±0.5834	6.345±0.4324	7.158±2.219	7.047±0.4366	7.338±0.5713	5.910±0.5031	8.333±0.9860
I	5.554±0.9125	7.028±0.9020	8.418±1.131	7.015±1.100	7.033±1.750	8.260±2.168	7.243±0.6404	7.663±1.328	6.478±1.662	8.735±0.8043
J	5.122±1.968	6.708±0.5741	6.513±1.257	6.060±0.2685^*	5.720±0.8029	5.948±0.8864	7.575±0.8563	6.630±0.8682	5.768±0.7062	8.170±1.141

The ^* indicates a significant difference compared with control group. ^*p < 0.05.

Additional Table 10 BUN contents in male rats in this research (X±SD, n≥2) (mmol/L).

Groups	Week
Groups	3	4	5	6	7	8	9	10	11	12
A	5.492±2.881	5.380±0.2358	5.323±0.4957	6.283±0.9962	5.723±0.1950	6.450±0.3751	6.823±0.4394	6.963±0.1762	5.910±0.9681	6.223±0.3262
B	4.628±0.8401	5.730±0.9235	6.833±1.036	6.655±0.2616	6.810±0.9192	6.775±0.3889	7.330±0.4243	7.360±0.6492	5.955±0.6859	6.825±0.6435
C	5.369±0.6171	4.858±0.5019	5.973±0.7466	6.260±0.2048	6.173±0.3874	6.900±0.8068	7.335±0.8214	7.095±0.8839	4.957±0.3014	7.170±0.6053
D	6.047±1.283	5.255±0.2855	5.625±0.4722	6.298±0.4373	5.718±0.3987	6.103±0.4924	6.828±0.2613	6.563±0.6678	4.858±0.2812	6.835±0.5344
E	5.369±1.402	6.693±0.0379^*	5.950±1.198	6.307±0.5839	5.770±0.4384	6.350±0	7.235±0.3323	6.375±0.5869	5.910±0	6.840±0.7284
F	5.862±1.111	6.423±0.7678	5.483±0.2984	6.727±1.572	6.570±0.4930	0.5000±0.2170	6.420±1.081	5.460±0.6172	4.650±0.4706	5.630±1.007
G	5.245±0.8155	5.388±0.6022	5.905±0.7221	5.753±0.3493	5.363±0.1517^*	6.103±0.5481	6.440±0.9122	7.130±0.3915	4.953±0.6676	6.197±0.6553
H	5.862±0.6492	6.603±0.1957^*	5.038±0.4279	6.423±1.193	6.683±0.5159^*	6.408±0.4511	5.868±0.7595	6.317±0.2759^*	5.503±1.154	6.447±0.4245
I	5.184±2.857	5.488±0.6896	5.253±0.5842	5.940±0.4228	5.788±0.5177	6.680±0.6707	6.035±0.9000	6.325±0.7196	5.413±0.1135	7.040±0.5446
J	4.443±0.2850	5.738±0.3759	6.118±0.6416	4.923±0.4446	5.595±0.8464	5.813±0.6530	6.428±0.7742	5.905±0.7537	4.635±0.6797	7.423±0.7570

The ^* indicates a significant difference compared with control group. ^*p < 0.05.

Additional Table 11 Scr contents in female rats in this research (X±SD, n≥2) (μmol/L).

Groups	Week
Groups	3	4	5	6	7	8	9	10	11	12
A	32.71±2.974	27.00±0.6083	29.07±2.750	27.60±3.780	28.00±1.556	33.90±1.697	32.20±2.687	131.5±41.58	107.9±48.08	29.65±3.182
B	23.79±8.921	31.23±4.379	30.60±2.165	33.35±4.876	32.08±2.395	35.83±1.394	31.50±3.780	64.20±37.50	89.50±49.65	36.43±4.280
C	26.76±6.424	31.85±5.786	30.75±4.800	31.40±4.032	30.23±2.547	36.35±6.784	31.50±3.780	84.27±40.84	107.3±37.40	33.83±6.086
D	32.71±4.856	31.25±2.164^*	31.13±3.442	28.93±2.830	27.63±2.272	31.28±4.145	30.18±3.877	84.80±54.26	122.3±7.946	32.37±4.761
E	30.48±2.847	28.20±3.311	29.23±1.522	36.88±3.911^*	30.05±1.912	34.58±3.267	29.13±4.430	106.8±44.85	90.75±43.46	29.95±3.155
F	30.48±8.880	33.85±9.056	30.20±4.544	31.10±1.445	29.75±1.670	30.27±2.797	27.15±2.339	103.8±44.56	71.03±30.78	28.28±1.928
G	35.68±40.01	32.20±2.762^*	27.50±5.032	25.23±3.707	28.13±5.262	28.33±4.045	29.87±6.072	74.70±41.16	106.7±19.52	29.10±4.662
H	15.61±9.825^*	28.68±2.077	27.68±2.300	30.55±2.290	29.15±3.908	30.38±1.928	47.25±11.95	82.97±27.10	121.1±13.97	34.13±2.654
I	23.05±5.079^*	28.60±3.451	28.50±3.767	29.65±4.914	28.90±4.185	32.73±6.557	31.75±6.903	93.45±42.75	83.60±56.95	35.95±7.123
J	17.84±8.754^*	26.58±3.207	28.63±2.664	30.80±3.831	29.00±1.645	28.68±0.6850^*	41.00±12.29	52.83±28.69	108.2±32.70	29.65±1.179

The ^* indicates a significant difference compared with control group. ^*p < 0.05.

Additional Table 12 Scr contents in male rats in this research (X±SD, n≥2) (μmol/L).

Groups	Week
Groups	3	4	5	6	7	8	9	10	11	12
A	18.83±3.434	22.17±3.156	23.20±2.951	22.73±1.790	20.63±0.6506	22.37±1.002	21.85±0.4950	66.60±59.39	123.3±14.54	56.15±48.30
B	23.79±6.424	25.73±3.634	26.53±3.620	30.05±1.061^*	26.30±0.1414^*	25.10±3.111	151.8±17.68^*	106.7±44.19	79.25±72.48	28.25±0.3536
C	20.07±1.487	23.20±1.738	24.08±3.758	23.25±1.996	23.35±2.709	23.97±1.721	61.30±31.89	87.10±56.21	53.55±27.37^*	27.67±4.631
D	24.53±10.69	23.73±0.8180	22.58±2.975	24.95±3.258	21.95±1.370	23.08±2.985	55.05±54.09	108.5±44.23	121.9±14.76	27.15±3.030
E	20.07±7.026	25.30±2.427	24.57±2.902	25.60±3.439	20.65±3.748	52.60±36.35	86.00±82.87	100.4±13.86	83.25±79.13	21.90±2.647
F	21.56±4.460	25.20±2.515	24.77±0.8327	24.73±2.026	21.50±3.964	25.50±3.466	141.3±49.07	77.90±11.40	77.80±14.96	25.90±0.3397
G	12.64±11.23	22.00±1.120	22.08±0.5500	22.73±1.511	21.00±1.225	24.30±1.802	77.15±58.31	72.97±60.15	86.55±19.45	23.47±0.9074
H	22.30±12.96	25.50±5.091	25.20±2.763	26.35±4.422	26.53±3.288^*	27.18±1.362^*	120.8±56.11	119.2±35.78	95.37±62.66	29.20±4.808
I	28.99±14.64	22.85±5.155	24.65±1.950	23.88±1.556	22.40±2.177	23.35±2.111	65.90±66.69	63.10±46.12	89.57±38.68	21.40±5.374
J	14.12±9.825	22.43±1.996	23.40±2.902	22.58±2.204	22.43±1.415	24.08±1.668	79.35±27.47^*	89.50±42.88	97.88±48.73	24.63±2.016

The ^* indicates a significant difference compared with control group. ^*p < 0.05.

Additional Table 13 HPCIII contents in rats in this research (X±SD, n≥2) (ng/mL).

Groups	Female			Male
Groups	4^th week	8^th week	12^th week	4^th week	8^th week	12^th week
A	ND	ND	ND	ND	ND	ND
B	ND	ND	ND	17.90±5.919	ND	ND
C	ND	ND	ND	ND	ND	ND
D	ND	ND	ND	ND	ND	ND
E	ND	ND	ND	ND	ND	ND
F	ND	ND	ND	ND	ND	ND
G	4.484±2.895	ND	ND	ND	ND	ND
H	ND	ND	ND	ND	ND	ND
I	ND	ND	ND	ND	ND	ND
J	ND	ND	ND	ND	ND	ND

ND: not detected.

Yanran He, Meizhen Song, Wangen Wang, Pei Lin, Yunfei Li, Wen Gu, Jie Yu, Ronghua Zhao. Chronic toxicity of both raw and processed Polygoni Multiflori Radix on rats[J]. Journal of Chinese Pharmaceutical Sciences, 2016, 25(1): 46-56.

References

[1] Chinese Pharmacopoeia Commission. Pharmacopoeia of the People’s Republic of China. Beijing: China Medical Science and Technology Press, 2010, 164-165.

[2] Niu, J.H. Chinese Journal of New Drugs and Clinical Remedies. 1996, 15, 382.

[3] Sun, J. Chin. J. Integr. Tradit. West. Med. Dig. 2002, 10, 292.

[4] Zhao, X.D. China J. Chin. Mater. Med. 2007, 32, 444.

[5] But, P.P.; Tomlinson, B.; Lee, K.L. Vet. Hum. Toxicol. 1996, 38, 280-282.

[6] Park, G.J.; Mann, S.P.; Ngu, M.C. J. Gastroenterol. Hepatol. 2001, 16, 115-117.

[7] Mazzanti, G.; Battinelli, L.; Daniele, C.; Mastroianni, C.M.; Lichtner, M.; Coletta, S. Ann. Intern. Med. 2004, 140, W30.

[8] Panis, B.; Wong, D.R.; Hooymans, P.M.; De, S.P.A.; Rosias, P.P. J. Pediatr. Gastroenterol. Nutr. 2005, 41, 256-258.

[9] Cardenas, A.; Restrepo, J.C.; Sierra, F.; Correa, G. J. Clin. Gastroenterol.2006, 40, 629-632.

[10] Yuen, M.F.; Tam, S.; Fung, J.; Wong, D.K.; Wong, B.C.; Lai, C.L. Aliment. Pharmacol. Ther. 2006, 24, 1179-1186.

[11] Min, H.J. J. Hepatol.2008, 47, S356.

[12] Hu, X.Q.; Yang, H.L.; Zhao, X.Q.; Miao, Y.X.; Geng, Z.Y. J. Shaanxi Coll. Tradit. Chin. Med. 2006, 29, 40-41.

[13] Hu, X.Q.; Geng, Z.Y.; Li, Q.L.; Fang, H.L.; Zhang, X.Q. Shaanxi Coll. J. Tradit. Chin. Med. 2007, 28, 1420-1421.

[14] Huang, W.; Zhang, Y.N.; Sun, R. Chin. J. Pharmacovigilance. 2010, 7, 705-707.

[15] Huang, W.; Zhang, Y.N.; Sun, R. Chin. J. Pharmacovigilance. 2011, 8, 193-197.

[16] Li, Y.; Xu, L.; Liu, R.N.; Chen, C.; Shi, L. J. Liaoning Univ. TCM. 2011, 13, 248-249.

[17] Li, W.X.; Zhang, Q.; Wang, G.R.; Lin, X.L.; Peng, Y.T. Guangming J. Chin. Med.2012, 27, 595-597.

[18] Wang, T.; Wang, J.Y.; Jang, Z.Z.; Zhou, Z.X.; Li, Y.Y.; Zhang, L. China J. Chin. Mater. Med. 2012, 37, 1445-1450.

[19] Wu, X.Q.; Chen, X.Z.; Huang, Q.C. Fitoterapia. 2012, 83, 469-475.

[20] Li, Q.; Zhao, K.J.; Zhao, Y.L. Glob. Tradit. Chin. Med. 2013, 6, 1-7.

[21] Zhang, C.; Zhang, R.C.; Sun, Z.X. J. Chin. Med. Mater. 2013, 36, 1416-1419.

[22] Yu, J.; Xie, J.; Zhao, R.H. Chin. Tradit. Herb. Drugs. 2010, 41, 11-15.

[23] Yu, J.; Xie, J.; Mao, X.J. J. Ethnopharmacol. 2011, 2011, 1291-1299.

[24] Law, M.R.; Wald, N.J.; Rudnicka, A.R. Br. Med. J. 2003, 326, 1423.25] Alberton, M.; Wu, P.; Druyts, E.; Briel, M.; Mills, E.J. Quarterly J. Med. 2012, 105, 145-157.

[

[26] National Health and Family Planning Commission of the People’s Republic of China. 2002, This article can be found online at http://www.nhfpc.gov.cn/zhuzhan/wsbmgz/ 201304/e33435ce0d894051b15490aa3219cdc4.shtml

[27] Ye, Q.H. Chin. J. Integr. Tradit. West. Med. 2012, 16, 732.

[28] Canadian Adverse Reaction Newsletter. 2003, This article can be found online at http://www.hc-sc.gc.ca/dhp-mps/medeff/bulletin/carn-bcei_v13n1-eng.php

[29] Medicines and Healthcare products Regulatory Agency. 2006, This article can be found online at http://www. mhra.gov.uk/NewsCentre/Pressreleases/CON2023635

[30] Medicines and Healthcare products Regulatory Agency. 2006, This article can be found online at http://www. mhra.gov.uk/NewsCentre/Pressreleases/CON2023590.

[31] Therapeutic Goods Administration. 2007, This article can be found online at http://www.tga.gov.au/pdf/archive/ cmec-minutes-61.pdf

[32] Therapeutic Goods Administration. 2008, This article can be found online at http://www.tga.gov.au/archive/labelling-rasml-notices-080414.htm#.U2ZW8bKBSDc

[33] Hu, X.Q.; Gao, J.; Li, Y.L.; Wang, L. Liaoning J. Tradit. Chin. Med.2011, 38, 988-990.

[34] Liu, Y.D.; Huang, X.X.; Wu, L.S.; Sun, D.M.; Lu, D.Y. Asia Pac. Tradit. Med. 2007, 71-72.