Effect of dl-3-n-butylphthalide on infarction volume in animal models of ischemic stroke: A meta-analysis

Table 1 Characteristics of the included studies

Study (yr)	Species / Age, Sex / Weight, Number (Control group / Experimental group)	Anesthetic	Model	Intervention dose administration method time point / duration	Measurement of infarction volume	Outcome index	P-values
Qin et al[8], 2018	SD rats/8 weeks	5 % chloral hydrate	tMCAO 2 h	90 mg/kg, daily	Cresyl violet	1 Infarction volume	P < 0.05
	Male/250-300 g	400 mg/kg	Reperfusion 7 d	Gavage	Image J	2 Ameliorate body weight loss	P < 0.05
	6/6	Intraperitoneally		Postoperative / 7 d		3 Improve neurological behavior scores	P < 0.05
						4 Reduce brain atrophy volume	P < 0.01
						5 Upregulate PTGIS, PTGES; downregulate TBXAS 1	P < 0.05
						6 Prevent REN,AGT, ACE 1, AGTR 1; upregulate RoA	P < 0.05
						7 Increase the diameter of middle cerebral artery	P < 0.05
Zhao et al[16], 2018	CD 1 mice / 10-12 wk	10 % chloral hydrate	pMCAO 24 h	120 mg/kg	2% TTC	1 Infarction volume	P < 0.05
	Male/27-30 g	35 mg/g				2 Improve neurological behavior scores	P < 0.05
	10/10	Intraperitoneally				3 Decrease the water content of brain	P < 0.05
						4 Decrease the permeability of blood-brain barrier	P < 0.05
						5 Decrease pinocytotic vesicles of capillary endothelial cells
						6 Downregulate MMP 9	P < 0.05
						7 Upregulate Claudin 5, VEGF, GFAP, Nrf 2 and HO 1	P < 0.05
Wang et al[18], 2018	SD rats/unknown	Unknown	tMCAO 2 h	1 mg/kg	2% TTC	1 Infarction volume	P > 0.05
	Male and female/250-280 g		Reperfusion 48 h	Intravenously	Image Pro Plus	2 Improve neurological behavior scores	P > 0.05
	8/12			postoperative/4 h and 24 h
Yan et al[17], 2017	SD rats/adult	Unknown	tMCAO 2 h	75 mg/kg, daily	2% TTC	1 Infarction volume	P < 0.01
	Male/180-220 g		Reperfusion 24 h	Gavage		2 Decrease the water content of brain	P < 0.05
	8/8			Preoperative/7 d		3 Decrease the permeability of blood-brain barrier	P < 0.01
						4 Decrease cell apoptosis
						5 Decrease ROS,cleaved caspase-3, p-p38; increase SOD	P < 0.01
						6 Decrease MDA, p-JNK	P < 0.05
Zhang et al[19], 2016	SD rats/unknown	Sodium pentobarbital	tMCAO 1 h	4.5 mg/kg	TTC	1 Infarction volume	P < 0.01
	Male/250-320 g	50 mg/kg	Reperfusion 24 h	Intraperitoneally		2 Improve neurological behavior scores	P < 0.001
	8/8			Postoperative/-		3 Decrease the water content of brain	P < 0.05
						4 Upregulate HGF; downreglate TLR4	P < 0.001
Yin et al[20], 2016	SD rats/unknown	Chloral hydrate	tMCAO 2 h	80 mg/kg, daily	2% TTC	1 Infarction volume	P < 0.001
	Male/280-320 g	300 mg/kg	Reperfusion 24 h	Gavage	Image Pro Plus	2 Improve neurological behavior scores	P < 0.05
	6/6	Intraperitoneally		preoperative/7 d		3 Decrease the water content of brain	P < 0.01
						4 Decrease MDA	P < 0.01
						5 Increase SOD	P < 0.05
						6 Increase GSH-Px	P < 0.001
Hua et al[21], 2015	SD rats/57-61 days	Chloral hydrate	tMCAO 2 h	60 mg/kg	2% TTC	1 Infarction volume	P < 0.05
	Male/250-280 g	300 mg/kg	Reperfusion 24 h	Gavage	Image Pro Plus	2 Improve neurological behavior scores	P > 0.05
	6/6	Intraperitoneally		Postoperative/-		3 Decrease the water content of brain	P < 0.01
						4 Decrease MDA; increase GSH, SOD, Nrf 2, Trx, Bcl-2	P < 0.01
						5 Decrease NF-κB p65; increase Txnip	P < 0.05
Lu et al[22], 2014	C57BL6/J/ dult	Isoflurane	tMCAO 0.75 h	10 mg/kg	Cresyl violet	1 Infarction volume	P < 0.05
	Male/ nknown	Initiated 3 %	Reperfusion 23 h	Intravenous	Image J	2 Improve neurological behavior scores	P < 0.05
	10/10	Maintained 1.5 %		Preoperative/-		3 Decrease MMP 9; increase TIMP 1	P < 0.01
						4 Increase SBP, p-ERK	P < 0.01
Wang et al[23], 2013	SD rats/unknown	Chloral hydrate	tMCAO 2 h	80 mg/kg, daily	TTC	1 Infarction volume	P < 0.05
	Male/280-320 g	300 mg/kg	Reperfusion 24 h	Gavage		2 Improve neurological behavior scores	P > 0.05
	6/6	Intraperitoneally		preoperative/7 d		3 Decrease the water content of brain	P > 0.05
Wang et al[24], 2013	SD rats/unknown	10 % chloral hydrate	tMCAO 2 h	80 mg/kg, daily	2% TTC	1 Infarction volume	P < 0.05
	Male/250 ± 20 g (n = 30)	3 mL/kg	Reperfusion 24 h	Gavage		2 Downregulate GRP78, CHOP	P < 0.05
	5/5	Intraperitoneally		Preoperative / 7 d
Pan et al[25], 2013	Wistar rats/unknown	10 % chloral hydrate	pMCAO 24 h	0.8 g/kg, daily	2% TTC	1 Infarction volume	P < 0.05
	Male/200-250 g	3 mL/kg		Gavage	Biosens Digitan Image	2 Decrease the water content of brain	P < 0.05
	8/8	Intraperitoneally		Preoperative/14 d		3 Decrease Smac, S100B	P < 0.05
Zhang et al[26], 2013	SD rats/unknown	6 % chloral hydrate	tMCAO 2 h	200 mg/kg	2% TTC	1 Infarction volume	P < 0.01
	Male/250 ± 20 g		Reperfusion 24 h	Intraperitoneally	Image Pro Plus	2 Improve neurological behavior scores	P < 0.01
	8/8	Intraperitoneally		Postoperative/-		3 Increase VEGF	P < 0.01
Wang et al[27], 2012	SD rats/unknown	chloral hydrate	tMCAO 2 h	90 mg/kg, daily	2% TTC	1 Infarction volume	P < 0.05
	Male/250-300 g	300 mg/kg	Reperfusion 3 d	Gavage		2 Improve neurological behavior scores	P < 0.05
	10/10	Intraperitoneally		Postoperative/3 d		3 Decrease the water content of brain	P < 0.05
						4 Decrease the ratio of TXB2 : 6-keto-PGF1α	P < 0.05
Wu et al[28], 2012	SD rats/unknown	Chloral Hydrate	tMCAO 2 h	80 mg/kg, daily	2% TTC	1 Infarction volume	P < 0.001
	Male/250-280 g	350 mg/kg	Reperfusion 24 h	Gavage		2 Improve neurological behavior scores	P < 0.001
	6/6	Intraperitoneally		Preoperative/7 d		3 Decrease the water content of brain	P > 0.05
						4 Decrease MDA; increase SOD	P < 0.001
Zhang et al[29], 2012	Wistar Kyoto rats/3 mo	Ketamine, xylazine	pMCAO 7 d	80 mg/kg, daily	2% TTC	1 Infarction volume	P < 0.01
	Male/unknown	75 mg/kg, 10 mg/kg		Gavage	Image Pro Plus
	6/6	Intraperitoneally		Postoperative/7 d
Zhao et al[30], 2012	SD rats/unknown	Chloral hydrate	tMCAO 2 h	50 mg/kg, daily	2% TTC	1 Infarction volume	P < 0.05
	Male/250-300 g	300 mg/kg	Reperfusion 3 d	Gavage		2 Improve neurological behavior scores	P < 0.05
	6/6	Intraperitoneally		Postoperative / 3 d
Li et al[31], 2010	129S2/Sv/adult	4 % chloral hydrate	pMCAO 24 h	100 mg/kg	2% TTC	1 Infarction volume	P < 0.05
	Male/20-25 g			Intraperitoneally		2 Decrease cleaved-caspase 3; caspase 9, p-JNK; p-p38	P < 0.05
	10/10	Intraperitoneally		Postoperative 1 h/-		3 Reduce mitochondrial release of cytochrome c and AIF	P < 0.05
Cao et al[32], 2009	SD rats/3-4 mo	10 % chloral hydrate	tMCAO 2 h	25 mg/kg, twice a day	TTC	1 Infarction volume	P < 0.01
	Male/280-350 g		Reperfusion 3 d	Gavage	Image Pro Plus	2 Improve neurological behavior scores	P < 0.05
	5/5	Intraperitoneally		Postoperative/3 d		3 Upregulate VEGF, bFGF	P < 0.05
Li et al[33], 2008	SD rats/3-4 mo	Unknown	pMCAO 3 d	25 mg/kg, twice a day	TTC	1 Infarction volume	P < 0.05
	Male/280-350 g			Gavage	Image Pro Plus	2 Improve neurological behavior scores	P < 0.05
	5/5			Postoperative/3 d		3 Upregulate VEGF, bFGF	P < 0.05
Zhang et al[34], 2006	SD rats/unknown	3 % isoflurane	tMCAO 2 h	10 mg/kg	4% TTC	1 Infarction volume	P < 0.001
	Male/270-330 g	Endotracheal intubation	Reperfusion 24 h	Intravenously	SPOT Biometrics	2 Improve neurological behavior scores	P < 0.01
	10/10			intraoperative / -
Lin et al[35], 1996	Wistar rats/unknown	Chloral hydrate	pMCAO 24 h	240 mg/kg	4% TTC	1 Infarction volume	P < 0.001
	Male/250-350 g			Gavage		2 Improve neurological behavior scores	P < 0.001
	8/8			Postoperative/-

PTGIS: Prostacyclin synthase; PTGES: Prostaglandin E synthase; TBXAS 1: Thromboxane A2 synthase 1; REN: Renin; AGT: Angiotensinogen; ACE 1: Angiotensin converting enzyme 1; AGTR 1: Angiotensin II receptor type 1; tMCAO: Transient middle cerebral artery occlusion; pMCAO: Permanent middle cerebral artery occlusion; TTC: 2,3,5-triphenyltetrazolium chloride; MMP 9: Matrix metallopeptidase 9; VEGF: Vascular endothelial growth factor; GFAP: Glial fibrillary acidic protein; Nrf 2: NF-E2-related factor 2; HO 1: Heme oxygenase 1; ROS: Reactive oxygen species; SOD: Superoxide dismutase; MDA: Malonaldehyde; HGF: Hepatocyte growth factor; TLR4: Toll like receptor 4; GSH-Px: Glutathione peroxidase; GSH: Glutathione; Trx: Thioredoxin; Txnip: Thioredoxin-interacting protein; TIMP 1: Tissue inhibitor of metalloproteinase 1; SBP: Spectrin breakdown product; GRP 78: Glucose-regulated protein 78; CHOP: C/EBP-homologous protein; Smac: Second mitochondria-derived activator of caspases; S100B: S100 calcium binding protein B; TXB2: Thromboxane B2; 6-keto-PGF1α: 6-keto-prostaglandin F1α; AIF: Apoptosis-inducing factor; bFGF: Basic fibroblast growth factor.

Table 2 Risk of bias of the included studies

Study (yr)	A	B	C	D	E	F	G	H	I	J	Score
Qin et al[8], 2018	√	√	√				√	√	√	√	7
Zhao et al[16], 2018	√	√	√						√	√	5
Wang et al[18], 2018	√		√	√				√	√		5
Yan et al[17], 2017	√			√					√	√	4
Zhang et al[19], 2016	√			√					√	√	4
Zhao et al[16], 2018	√	√	√						√	√	5
Yin et al[20], 2016	√	√	√					√	√	√	6
Hua et al[21], 2015	√	√	√	√				√	√	√	7
Lu et al[22], 2014	√	√	√	√				√	√	√	7
Wang et al[23], 2013	√	√	√						√	√	5
Wang et al[24], 2013	√	√	√					√		√	5
Pan et al[25], 2013	√	√	√					√		√	5
Zhang et al[26], 2013	√	√	√						√	√	5
Wang et al[27], 2012	√	√	√						√	√	5
Wu et al[28], 2012	√	√	√						√	√	5
Zhang et al[29], 2012	√	√	√	√				√	√		6
Zhao et al[30], 2012	√	√	√	√				√	√	√	7
Li et al[31], 2010	√								√	√	3
Cao et al[32], 2009	√	√	√	√			√	√		√	7
Li et al[33], 2008	√	√	√	√				√		√	6
Zhang et al[34], 2006	√	√	√	√					√	√	6
Lin et al[35], 1996	√								√	√	3

A: Baseline characteristics; B: Allocation concealment; C: Sequence generation; D: Random housing; E: Blind feeding; F: Random outcome assessment; G: Result evaluator blind; H: No incomplete outcome data; I: No selective outcome reporting; J: No other sources of bias.