Study on anxiety-like behavior of mice with late-stage infection of Schistosoma japonicum

doi:10.12140/j.issn.1000-7423.2022.05.009

Abstract

Abstract:

Objective To observe the behavioral characteristics of mice with late-stage schistosomiasis japonica and explore the relationship between late-stage infection of Schistosoma japonicum and anxiety. Methods Thirty 6-week-old male C57BL/6J mice were randomly divided into the infected group (n = 16) and the uninfected group (n = 14). The infected group was infected via abdominal skin contact with 20 cercariae of both sexes, and the uninfected group was treated with dechlorinated water without cercariae. Mice in both of groups were administered with praziquantel by gavage at 8 weeks post-infection. Additional 7 male APP/PS1 double transgenic mice (Alzheimer’s disease model) at 6 weeks of age were used as positive control (transgenic group) without any treatment. Three groups of mice were reared in the same environment until they were 10 months old, to observe their behavioral changes in an open-field test (OFT) and elevated-plus maze test (EPM). In OFT, the total distance, the travelling distance in the central zone, the percentage of travelling distance in the centrol zone, the number of locomotor activity, the duration of locomotor activity, the time percentage spent in the centrol zone, the mean speed, the mean speed in the centrol zone were measured, and in EPM, the locomotor distance, retention time and the number of entry in the open arms/close arms/central zone, and the percentage of locomotor distance, retention time and number of entry in the open arms were recorded to evaluate the anxiety-like behavior of the three mice groups. The mice of all groups were euthanized, and the livers were collected to observe gross morphology. The liver tissue sections were stained using hematoxylin & eosin (HE) and Masson’s trichrome stain for microscopic observation of pathological changes and collagen fiber hyperplasia. The content of liver hydroxyproline which is the specific amino acid constituent in collagen, was determined. The mRNA relative transcription levels of α-smooth muscle actin (α-SMA), transforming growth factor-β1 (TGF-β1), collagen Ⅰ(ColⅠ) and ColⅢ in the liver tissues of each group were detected by fluorescence quantitative PCR. SPSS 21.0 software was used for statistical analysis. Results The appaearance color of the mice livers from the infected group was darker compared to the uninfected group. There was a large number of eggs were deposited in the liver tissue, with surrounded infiltrating inflammatory cells and collagen fibres hyperplasia. The liver hydroxyproline content of the infected group (0.194 ± 0.040) μg/mg was significantly higher than that of the uninfected group (0.122 ± 0.016) μg/mg and transgenic group (0.124 ± 0.004) μg/mg (t = 0.79, 0.86, P < 0.05), with no significant difference between the uninfected and transgenic group. The mRNA expression levels of Col Ⅰ, α-SMA, TGF-β1, Col Ⅲ in the liver of the infected group were 3.55 ± 1.76, 2.01 ± 0.66, 2.21 ± 0.64, 4.51 ± 2.32,which is not significantly higher than those of the uninfected group (t = 0.76, 0.59, 0.17, 0.40,P > 0.05). The infected group’s activity time (346.77 ± 181.03) in OFT, the locomotor distance in the open arms (206.31 ± 282.52) mm, the percentage of locomotor distance in the open arms (7.51 ± 9.90)%, the number of entry in the open arms, closed arms and central zone (1.32 ± 1.51, 3.44 ± 3.10, 4.53 ± 4.21)(P < 0.01) in EPM were significantly increased compared to the uninfected group (t = 1.61, 2.24, 1.66, 1.83, 2.35, 2.22, P < 0.05 or P < 0.01), with no significant differences between the two groups for the other indexes. The infected group’s the number of locomotor activity (123.68 ± 50.23) in OFT was significantly decreased compared to the transgenic group (P < 0.05), but there was no significant difference between the two groups for other indexes of OFT. The infected group showed shorter retention time in the closed arm (232.85 ± 88.82) s than that in the transgenic group (295.38 ± 9.88）s(t = -2.81,P < 0.01), and higher values of the rest of the indexes in EPM than those in the transgenic group (P < 0.05 or P < 0.01). Conclusion Mice with late-stage of S. japonica infection did not show more serious level of anxiety than the uninfected mice, however, the anxiety extent presented a significant reducing-tendency, suggesting that the pathological process of S. japonica infection may not be directly correlated to the occurrence and development of anxiety.

Key words: Late-stage infection of Schistosoma japonicum, Open-field test, Elevated-plus maze test, Anxiety

CLC Number:

R532.21

TANG Xian-shi, JI Wen-xiang, XIONG Chun-rong, ZHOU Yong-hua, XU Yong-liang, TONG De-sheng. Study on anxiety-like behavior of mice with late-stage infection of Schistosoma japonicum[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2022, 40(5): 622-628.

Figures/Tables 5

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References 24

[1]	LoVerde PT. Schistosomiasis[J]. Adv Exp Med Biol, 2019, 1154: 45-70. doi: 10.1007/978-3-030-18616-6_3 pmid: 31297759
[2]	Collins C,, Xu J,, Tang SL. Schistosomiasis control and the health system in P. R. China[J]. Infect Dis Poverty, 2012, 1: 8. doi: 10.1186/2049-9957-1-8
[3]	Zhang LJ,, Xu ZM,, Yang F, et al. Endemic status of schistosomiasis in People’s Republic of China in 2020[J]. Chin J Schisto Control, 2021, 33(3): 225-233. (in Chinese)
	( 张利娟,, 徐志敏,, 杨帆, 等. 2020年全国血吸虫病疫情通报[J]. 中国血吸虫病防治杂志, 2021, 33(3): 225-233.)
[4]	Deng Y. The study on disease burden of advanced schistosomiasis and the quality of life of patients[D]. Beijing: Chinese Center for Disease Control and Prevention, 2008: 6-16. (in Chinese)
	( 邓瑶. 晚期血吸虫病疾病负担与患者生命质量研究[D]. 北京: 中国疾病预防控制中心, 2008: 6-16.)
[5]	Hua HY,, You H,, Zhang Y, et al. Evaluation of quality of life in advanced schistosomiasis patients in Jiangsu Province[J]. Chin J Schisto Control, 2010, 22(6): 562-566. (in Chinese)
	( 华海涌,, 尤华,, 张燕, 等. 江苏省晚期血吸虫病患者生命质量评估[J]. 中国血吸虫病防治杂志, 2010, 22(6): 562-566.)
[6]	Jiang CG,, Chen SJ,, Li SM, et al. Study on economic and social burden caused by advanced schistosomiasis in Jurong City[J]. Chin J Schisto Control, 2010, 22(6): 616-618. (in Chinese)
	( 江成功,, 陈世军,, 李水明, 等. 句容市晚期血吸虫病经济及社会负担研究[J]. 中国血吸虫病防治杂志, 2010, 22(6): 616-618.)
[7]	Li YT. Study on disease burden of advanced schistosomiasis Japonica in China[D]. Beijing: Chinese Center for Disease Control and Prevention, 2019: 6-14. (in Chinese)
	( 李伊婷. 中国晚期血吸虫病疾病负担研究[D]. 北京: 中国疾病预防控制中心, 2019: 6-14.)
[8]	Nie YX. Depression prevalence and quality of life of patients with advanced schistosomiasis[J]. Chin J Schisto Control, 2011, 23(5): 579-581. (in Chinese)
	( 聂永新. 晚期血吸虫病患者抑郁症发生率及生活质量调查[J]. 中国血吸虫病防治杂志, 2011, 23(5): 579-581.)
[9]	Zhou RH,, Pan J,, Liu KF, et al. Quality of life and influencing factors in patients with advanced schistosomiasis complicated with depression[J]. J Nurs, 2014, 21(16): 65-68. (in Chinese)
	( 周瑞红,, 潘洁,, 刘科丰, 等. 晚期血吸虫病合并抑郁患者生活质量及影响因素研究[J]. 护理学报, 2014, 21(16): 65-68.)
[10]	Pan J,, Zhou RH,, Liu KF, et al. Investigation on anxiety, depression and life quality of hospitalized patients with advanced schistosomiasis and related influencing factors[J]. J Trop Med, 2014, 14(8): 1082-1085, 1092. (in Chinese)
	( 潘洁,, 周瑞红,, 刘科丰, 等. 晚期血吸虫病住院患者焦虑抑郁与生活质量状况及影响因素研究[J]. 热带医学杂志, 2014, 14(8): 1082-1085, 1092.)
[11]	Liu JY,, Yang DD. Effects of artesunate on the expression of type ⅰ, ⅲ, ⅳ collagen and TGF-β1 in liver tissue of mice with hepatic fibrosis[J]. Jilin J Tradit Chin Med, 2009, 29(5): 447-448. (in Chinese)
	( 刘金元,, 杨冬娣. 青蒿琥酯对肝纤维化小鼠肝组织中Ⅰ、Ⅲ、Ⅳ型胶原及TGF-β1表达的影响[J]. 吉林中医药, 2009, 29(5): 447-448.)
[12]	Zhou YH,, Xu C,, Yang YY, et al. Effect of artesunate on expression of heat shock protein 47 in mice with liver fibrosis induced by Schistosoma japonicum[J]. Chin J Parasitol Parasit Dis, 2019, 37(2): 115-120, 126. (in Chinese)
	( 周永华,, 徐辰,, 杨莹莹, 等. 青蒿琥酯对日本血吸虫诱导的早期肝纤维化小鼠热休克蛋白47表达影响的研究[J]. 中国寄生虫学与寄生虫病杂志, 2019, 37(2): 115-120, 126.)
[13]	Xu X,, Fu JB,, Yao Z, et al. Therapeutical effect of dihydroartemisinin on bile duct ligation-induced liver fibrosis in rats and its mechanism[J]. J Nanjing Univ Tradit Chin Med, 2017, 33(4): 376-381. (in Chinese)
	( 许霞,, 付金柏,, 姚真, 等. 双氢青蒿素对胆管结扎致大鼠肝纤维化的治疗作用研究[J]. 南京中医药大学学报, 2017, 33(4): 376-381.)
[14]	Kraeuter AK,, Guest PC,, Sarnyai Z. The open field test for measuring locomotor activity and anxiety-like behavior[J]. Methods Mol Biol, 2019, 1916: 99-103.
[15]	Liu Y,, Yao RZ,, Shi HZ, et al. Detection of spontaneous movement, exploration behavior and anxiety in ICR mice and establishment of reference range of corresponding indicators[J]. J Heilongjiang Bayi Agric Univ, 2020, 32(1): 29-35. (in Chinese)
	( 刘洋,, 姚睿智,, 史宏昭, 等. ICR小鼠自发运动、探索行为和焦虑情绪的检测及其参考值的确立[J]. 黑龙江八一农垦大学学报, 2020, 32(1): 29-35.)
[16]	Seibenhener ML,, Wooten MC. Use of the open field maze to measure locomotor and anxiety-like behavior in mice[J]. J Vis Exp, 2015(96): e52434.
[17]	Xin ZM,, Chen CJ. Study on anxiety-like behaviors of Zucker diabetic fatty model rats[J]. Mod Prev Med, 2019, 46(22): 4166-4169. (in Chinese)
	( 辛志明,, 陈超杰. Zucker糖尿病肥胖模型大鼠的焦虑样行为学研究[J]. 现代预防医学, 2019, 46(22): 4166-4169.)
[18]	Kraeuter AK,, Guest PC,, Sarnyai Z. The elevated plus maze test for measuring anxiety-like behavior in rodents[J]. Methods Mol Biol, 2019, 1916: 69-74.
[19]	Tang XS,, Zhao S,, Xiong CR, et al. Therapeutic effect of arachidonic acid (ARA) on schistosomiasis japonicum in mice[J]. Chin J Zoonoses, 2021, 37(8): 722-727. (in Chinese)
	( 汤宪时,, 赵松,, 熊春蓉, 等. 花生四烯酸对小鼠日本血吸虫病治疗效果观察[J]. 中国人兽共患病学报, 2021, 37(8): 722-727.)
[20]	Kuring JK,, Mathias JL,, Ward L. Prevalence of depression, anxiety and PTSD in people with dementia: a systematic review and meta-analysis[J]. Neuropsychol Rev, 2018, 28(4): 393-416. doi: 10.1007/s11065-018-9396-2 pmid: 30536144
[21]	Leontjevas R,, van Hooren S,, Mulders A. The montgomery-asberg depression rating scale and the cornell scale for depression in dementia: a validation study with patients exhibiting early-onset dementia[J]. Am J Geriatr Psychiatry, 2009, 17(1): 56-64. doi: 10.1097/JGP.0b013e31818b4111 pmid: 19092312
[22]	Bennett S,, Thomas AJ. Depression and dementia: cause, consequence or coincidence?[J]. Maturitas, 2014, 79(2): 184-190. doi: 10.1016/j.maturitas.2014.05.009 pmid: 24931304
[23]	Qu XS,, Shi H,, Cao XL, et al. Effects of adiponectin on the anxiety and memory impairment in triple transgenic Alzheimer’s disease model mice[J]. Chin J Appl Physiol, 2017, 33(5): 405-409. (in Chinese)
	( 曲雪松,, 石辉,, 曹秀丽, 等. 脂联素改善阿尔茨海默病模型小鼠焦虑情绪及工作记忆[J]. 中国应用生理学杂志, 2017, 33(5): 405-409.)
[24]	Bai Y. The neuroprotective effects and mechanism of adiponectin alleviates the anxiety- and depression-like behaviors in triple transgenic Alzheimer’s disease model mice[D]. Taiyuan: Shanxi Medical University, 2019: 15-22. (in Chinese)
	( 白羽. 脂联素改善三转基因阿尔茨海默病模型小鼠焦虑和抑郁样行为的神经保护作用及机制研究[D]. 太原: 山西医科大学, 2019: 15-22.)

指标 Index	感染组 Infection group	未感染组 Uninfection group	转基因组 Transgenic group
总路程/mm Total distance/mm	33 476.8 ± 71 190.44	12 719.55 ± 6 451.56	21 710.23 ± 11 544.04^c
中央区路程/mm Travelling distance in the central zone/mm	4 637.04 ± 10 362.40	1 569.83 ± 1 337.72	3 016.18 ± 2 335.31^c
中央路程百分比/% Percentage of travelling distance in the centrol zone/%	12.34 ± 4.37	11.00 ± 5.54	12.81 ± 5.25
活动次数/次 Number of locomotor activity/times	123.68 ± 50.23^b	119.14 ± 48.37	147.90 ± 41.17^c
活动时间/s Duration of locomotor activity/s	346.77 ± 181.03^a	273.53 ± 113.64	378.42 ± 128.24^c
中央时间百分比/% Time percentage spent in centrol zone/%	54.83 ± 49.67	52.43 ± 68.37	60.71 ± 50.88
平均速度/mm·s^-1 Mean speed/mm·s^-1	37.15 ± 79.01	14.11 ± 7.16	24.09 ± 12.81^c
中央平均速度/mm·s^-1 Mean speed in the centrol zone/mm·s^-1	79.29 ± 88.68	61.11 ± 57.44	71.14 ± 43.51

指标 Index	感染组 Infection group	未感染组 Uninfection group	转基因组 Transgenic group
开臂内运动路程/mm Locomotor distance in the open arms/mm	206.31 ± 282.52^ab	47.43 ± 105.82	28.49 ± 73.41
闭臂内运动路程/mm Locomotor distance in the closed arms/mm	2 304.63 ± 1704.01^b	3 959.01 ± 17117.29	1 512.28 ± 914.86
中央运动路程/mm Locomotor distance in the central zone/mm	120.48 ± 112.95^b	80.51 ± 140.23	14.26 ± 24.78^c
开臂内运动路程百分比/% Percentage of locomotor distance in the open arms/%	7.51 ± 9.90^ab	3.39 ± 10.02	1.36 ± 3.53
开臂进入次数/次 Number of entry in the open arms/times	1.32 ± 1.51^ab	0.63 ± 1.01	0.12 ± 0.33^c
闭臂进入次数/次 Number of entry in the close arms/times	3.44 ± 3.10^ab	1.60 ± 1.75	0.83 ± 1.04
中央进入次数/次 Number of entry in the central zone/times	4.53 ± 4.21^ab	2.19 ± 2.57	0.95 ± 1.23^c
开臂进入次数百分比/% Percentage of number of entry in the open arms/%	20.31 ± 20.71^b	16.05 ± 23.59	4.30 ± 12.45^c
开臂内停留时间/s Retention time in the open arms/s	25.80 ± 35.99^b	15.60 ± 48.22	3.44 ± 7.86
闭臂内停留时间/s Retention time in the closed arms/s	232.85 ± 88.82^b	262.27 ± 74.93	295.38 ± 9.88^c
中央停留时间/s Retention time in the central dwell time/s	42.33 ± 71.66^b	23.12 ± 52.75	2.17 ± 3.45
开臂内停留时间百分比/% Percentage of retention time in the open arms/%	8.57 ± 11.95^b	5.18 ± 16.02	1.14 ± 2.61