Effects of drug serum of anti-fibrosis I herbal compound on calcium in hepatic stellate cell and its molecular mechanism

Table 1 Factor-level for orthogonal design.

	Factor
Level	CCl4(mmol/L)	Durg serum(mL/L)	TGF-β1 Ab(mg/mL)	Time1(h)	Turn
1	5	5	5	0.5	Adding CCl4 firstly
2	10	10	10	2	Adding durg serum/TGF-β1 Ab firstly
3	15	20	20	4

¹Time is the interval time between CCl₄ and TGF-β₁ antibody or CCl₄ and anti-fibrosis I drug serum.

Table 2 Plan of orthogonal trial and the result of Ca²⁺ assay (mean±SD).

Trial	CCl4(mmol/L)	Drug serum(mL/L)	TGF-β1 Ab(mg/mL)	Time(h)	Turn	Fluorescence intensity
						TGF-β1 Ab	Drug serum
1	5	5	5	0.5	1	81.03±7.36	78.25±6.84
2	15	20	20	0.5	1	126.52±13.54	109.38±15.63
3	5	10	10	2	1	75.84±6.81	72.54±9.52
4	10	20	20	2.0	1	106.46±10.23	88.65±10.38
5	10	5	5	4	1	117.65±9.26	102.13±12.45
6	15	10	10	4	1	138.24±12.57	113.36±11.82
7	10	10	10	0.5	2	94.53±8.67	84.47±8.23
8	15	10	10	0.5	2	128.67±11.56	96.48±9.65
9	15	5	5	2	2	138.21±14.83	118.65±12.34
10	5	20	20	2	2	62.17±6.58	50.16±6.48
11	10	5	5	4.0	2	114.28±8.37	92.54±8.75
12	5	20	20	4	2	64.23±5.26	46.23±3.64

Table 3 Results of analysis of variance for orthogonal design.

Sources of variation	Sum of squares of deviation from mean	Degree of freedom	Variance	F	P
Variance analysis of TGF-β1 Ab
CCl4	6933.936	2	3466.968	392.639	0
TGF-b1 Ab	378.054	2	189.027	21.408	0.007
Time	67.786	2	33.893	3.838	0.117
Turn	158.777	1	158.777	17.982	0.013
Error	35.320	4 8.	830
Variance analysis of drug serum
CCl4	3969.848	2	1984.924	139.437	0
Drug serum	656.001	2	328.000	23.041	0.006
Time	55.274	2	27.637	1.941	0.257
Turn	478.551	1	478.551	33.617	0.004
Error	56.941	4	14.235

Table 4 Range analysis for orthogonal trial.

Level	Drug serum				TGF-β1 Ab
	CCl4	Drug serum	Time	Turn	CCl4	TGF-β1 Ab	Time	Turn
1	61.80	97.89	92.15	94.05	70.82	112.79	107.69	107.62
2	91.95	91.71	82.50	81.42	108.23	109.32	95.67	100.35
3	109.47	73.61	88.57		132.91	89.85	108.6
R	47.67	24.28	9.65	12.63	37.41	19.47	12.93	7.27

Table 5 Change of intracellular Ca²⁺ in hepatic stellate cell treated with different methods (mean±SD).

Group Methods of treatment		Fluorescence intensity
1	CCl4+HSC+drug serum	109.38±15.63
2	CCl4+HSC+TGF-β1 Ab	126.52±13.54
3	CCl4+HSC+ drug serum +TGF-β1 Ab	103.40±9.87
4	CCl4+HSC+TGF-β1 Ab+ drug serum	93.7796.33