加拿大一枝黄花石油醚萃取物对福寿螺肝脏的影响

doi:10.3864/j.issn.0578-1752.2019.15.007

摘要/Abstract

摘要：

【目的】在前期研究的基础上深入探讨加拿大一枝黄花石油醚萃取物(petroleum ether extract of Solidago canadensis,PEEE)对福寿螺（Pomacea canaliculata）肝脏的影响,为进一步有效控制福寿螺对农业生产的危害、实现恶性外来入侵植物加拿大一枝黄花的资源化利用提供依据。【方法】以去氯自来水作为对照组,0.11、0.18和0.29 mg?mL ^-1的PEEE悬浮液作为试验组。每组随机放入大小、活力一致的成年福寿螺10只,用一层纱布和橡皮筋封住烧杯口,防止福寿螺逃逸。在室温(23—27℃)条件下,处理48 h,每组3次重复。48 h后,分别随机挑选去氯自来水、0.11、0.18和0.29 mg?mL ^-1浓度处理下的福寿螺,敲破螺壳,取出肝脏,使用扫描电镜和透射电镜观察肝脏组织的超微结构。经0.29 mg?mL ^-1浓度处理后的福寿螺肝脏组织冻干后用于代谢组学研究。【结果】扫描电镜观察结果表明,经不同浓度的PEEE浸泡处理福寿螺48 h后,福寿螺肝脏表面均受到损伤,其损伤程度与PEEE处理液的浓度呈正相关,高浓度的PEEE可致使肝脏萎缩失去弹性,皱褶凸起被侵蚀,体表破损,出现絮状物,质地松散。透射电镜观察结果表明,经不同浓度的PEEE浸泡处理福寿螺48 h后,福寿螺肝细胞损伤明显,高浓度的PEEE使福寿螺肝细胞粗面内质网及线粒体数量减少,粗面内质网出现肿胀、断裂和颗粒脱落等现象;线粒体表现为肿胀,形状不规则。代谢组学的试验结果表明,经PEEE处理后,福寿螺肝脏中60种代谢物含量发生变化,多个代谢通路受到影响,富集程度最高的两条通路分别为糖鞘脂代谢和棕榈酰辅酶A进入线粒体基质的通路。【结论】加拿大一枝黄花的石油醚萃取物对福寿螺肝脏细胞结构和生理功能有巨大的损伤破坏作用,因而具有较强的灭螺活性,有望开发成为新型的天然灭螺药物。

关键词: 加拿大一枝黄花, 石油醚萃取物, 福寿螺, 肝脏, 扫描电镜, 透射电镜, 代谢组学

Abstract:

【Objective】 On the basis of previous studies, the effect of petroleum ether extract of Solidago canadensis (PEEE) on the liver of Pomacea canaliculata was discussed in depth to provide a basis for further effective control the harm of P. canaliculata to agricultural production and realizing the resource utilization of the malignant alien invasive plant S. canadensis. 【Method】Chlorinated tap water was used as the control group, and PEEE suspensions of 0.11, 0.18 and 0.29 mg?mL ^-1 were used as the experimental groups. In each group, 10 adult P. canaliculata of the same size and vigor were randomly placed, and the beaker was sealed with a layer of gauze and rubber band to prevent the P. canaliculata from escaping. At room temperature (23-27℃), the treatment was carried out for 48 h with 3 repetitions in each group. After 48 h, the P. canaliculata treated with dechlorinated tap water, 0.11, 0.18 and 0.29 mg?mL ^-1 were randomly selected, the shells of P. canaliculata were broken and the liver was taken out, and the ultrastructure of liver tissue was observed by scanning electron microscopy and transmission electron microscopy. The liver tissue of P. canaliculata treated with 0.29 mg?mL ^-1 was lyophilized for metabonomics study. 【Result】The results of scanning electron microscopy showed that the liver surface of P. canaliculata was damaged after soaking for 48 h with different concentrations of PEEE, and the degree of damage was positively related to the concentration of PEEE treatment solution. High concentrations of PEEE could cause liver atrophy and loss of elasticity, wrinkle bulge erosion, body surface damage, flocs and loose texture. The results of transmission electron microscopy showed that the liver cells of P. canaliculata were damaged obviously after soaking for 48 h with different concentrations of PEEE. The number of rough endoplasmic reticulum and mitochondria in liver cells of P. canaliculata decreased with high concentrations of PEEE, and the rough endoplasmic reticulum appeared swelling, breaking and degranulation. Mitochondria showed swelling and irregular shape. The results of metabonomic experiments showed that after PEEE treatment, the content of 60 metabolites in the liver of P. canaliculata changed, and several metabolic pathways were affected. The two pathways with the highest enrichment were glycosphingolipid metabolism and palmitoyl-CoA entering mitochondrial matrix pathway. 【Conclusion】PEEE of S. canadensis has great damage and destruction to the liver cells of P. canaliculata, so it has strong molluscicidal activity and is expected to be developed into a new natural molluscicidal drug with great potential development and application value.

Key words: Solidago canadensis, petroleum ether extract, Pomacea canaliculata, liver, scanning electron microscope, transmission electron microscope, metabonomics

李帅岚,沈校,邹峥嵘. 加拿大一枝黄花石油醚萃取物对福寿螺肝脏的影响[J]. 中国农业科学, 2019, 52(15): 2624-2635.

LI ShuaiLan,SHEN Xiao,ZOU ZhengRong. Effect of Petroleum Ether Extract from Solidago canadensis on Liver of Pomacea canaliculata[J]. Scientia Agricultura Sinica, 2019, 52(15): 2624-2635.

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时间 Time (h)	A%	B%	液相曲线Curve
0	99	1	起始Initial
1.0	99	1	6
1.5	80	20	6
11.0	1	99	6
11.5	2	98	6
14.0	99	1	1

序号Number	名称 Name	m/z	RT (min)	变化Change
1	(2S,3R,4E)-2-Ammonio-3-hydroxy-4-octadecen-1-yl 2-(trimethylammonio)ethyl phosphate	448.34	7.61	↓
2	12-epi-scalaradial	446.33	7.18	↓
3	(3alpha,5beta)-3,17,21-Trihydroxypregnan-20-one	368.28	6.07	↓
4	(2alpha,3beta,5alpha,25S)-Spirostan-2,3-diol	450.36	7.92	↓
5	1-Myristoyl-sn-glycero-3-phosphocholine	468.31	6.88	↓
6	1-Octadecanoyl-sn-glycero-3-phosphocholine	546.36	7.97	↓
7	(3alpha,5beta,7alpha,12alpha,24R)-Cholestane-3,7,12,24,26,27-hexol	486.38	7.96	↓
8	20-Hydroxyrhodopin	573.47	11.51	↓
9	axisothiocyanate-3	246.17	2.38	↑
10	Hexadecan-1-ol	260.29	2.85	↑
11	Palmitoylcarnitine	400.34	7.51	↑
12	Citicoline	489.12	0.55	↑
13	3-Indoleacrylic acid	188.07	2.24	↑
14	Trp-Ala-Arg	432.24	1.64	↑
15	trans-2-Hydroxycinnamic acid	182.08	1.40	↑
16	4-Hydroxyretinoic acid	317.21	8.42	↑
17	(2S)-2-[(3aS,5E,9E,13S,14E,16aR)-2,13-Dihydroxy-3a,6,10,14- tetramethyl-3- oxo- 3,3a,4,7,8,11,12,13,16,16a-decahydrocyclopenta[15]annulen-1-yl]propyl acetate	462.32	8.42	↑
18	(5Z,8Z,11Z,14Z)-16-[(2R,3S)-3-Ethyl-2-oxiranyl]-5,8,11,14-hexadecatetraenoic acid	301.22	6.88	↑
19	9-Oxo-prosta-10,12Z,14E-trienoic acid	319.23	7.07	↑
20	2,17beta-Dihydroxy-17-methylandrosta-1,4-dien-3-one	317.21	7.73	↑
21	18-Hydroxyretinoic acid	317.21	7.22	↑
22	Telecinobufagin	425.23	7.73	↑
23	(+)-Ophiobolin A	423.25	9.45	↑
24	Remiprostol	439.25	8.05	↑
25	Chivosazole C	855.50	8.05	↑
26	2-O-Sulfo-L-idopyranuronic acid	275.01	1.33	↑
27	Ricinoleic acid methyl ester	313.27	7.57	↑
28	3,4-Dihydroxyphenylalanine, butyl ester	507.26	7.59	↑
29	N-Acetyl-b-D-glucosamine	244.08	0.59	↑
30	L-alpha-Glycerylphosphorylcholine	258.11	0.52	↑