梅花草提取物对幽门螺杆菌小鼠胃黏膜定植的影响及其机制

doi:10.13481/j.1671-587X.20220505

Abstract

Abstract:

Objective To explore the effects of Parnassia palustris Linn extracts on Helicobacter pylori （Hp） eradication and gastric mucosa， and to clarify its antibacterial and anti-inflammatory mechanisms in vitro and in vivo. Methods The six-week old male Kunming mice were randomly divided into blank control group， normal saline group， Hp group， triple group， quadruple group， Parnassia palustris Linn group， triple + Parnassia palustris Linn group， and quadruple + Parnassia palustris Linn group. The minimal inhibitory concentration （MIC） of Parnassia palustris Linn extracts on Hp SS1 standard strains in vitro and the expression levels of Hp colonization- and adhesion-related gene mRNA were detected by real-time fluorescence quantitative （RT-qPCR） method. Animal experiments were used to evaluate the eradication rate of Hp by different methods， and enzyme linked immunosorbent assay（ELISA） method was used to detect the serum levels of inflammatory factors such as interleukin-2 （IL-2）， interleukin-8 （IL-8） and tumor necrosis factor alpha （TNF-α） of the mice in various groups. Rapid urease test （RUT） was used to detect the infection rate of Hp in stomach tissue， HE staining and transmission electron microscope were used to detect the pathomorphology and ultrastructures of stomach tissue. Results The MIC of Parnassia palustris Linn was 10 g·L^－1.The RT-qPCR results showed that compared with Hp group，the expression levels of Bab A， Cag A and Vac A mRNA in Parnassia palustris Linn group were decreased （P<0.05 or P<0.01）.The immunohistochemistry results showed that the part of gastric mucosa had erosion and ulcers，obvious bleeding and conggestion，and positive Hp infection of the mice in Hp group； the symptoms of gastric mucosal bleeding and ulcers in Parnassia palustris Linn group were improved， and Hp infection was mostly negative.The Rut results showed that compared with Hp group，the infection rate of Hp of the mice in Parnassia palustris Linn group was decreased（P<0.05），and the infection rates of the mice in triple group and quadruple group were decreased（P<0.05）；compared with Parnassia palustris Linn group，the eradication rates of Hp of the mice in triple group and quadruple group had no significant differences（P>0.05）； compared with triple group or quadruple group， the eradication rates of Hp of the mice in triple+Parnassia palustris Linn group and quadruple+Parnassia palustric Linn group were increased（P<0.01）. The gastric mucosa was apparently repaired， and the structures of mitochondria and microvilli of gastric mucosal epithelial cells were intact of the mice in triple+Parnassia palustris Linn group and quadruple+Parnassia palustris Linn group.Compared with Hp group，the levels of IL-2 in serum of the mice in triple， quadruple，Parnassia palustris Linn， triple+Parnassia palustris Linn and quadruple+Parnassia palustris Linn groups were increased（P<0.05）， and the levels of serum IL-8 and TNF-α were decreased（P<0.05 or P<0.01）. Conclusion Parnassia palustris Linn extracts can effectively eradicate Hp and protect gastric mucosa， and its mechanism is related to decreasing the levels of IL-2，IL-8 and TNF-α and protecting the integrity of structures of gastric mucosa subcellular organelle after Hp infection.

Key words: Parnassia palustris Linn, Helicobacter pylori, Colonization, Eradication rate, Transmission electron microscope, Inflammatory factors

CLC Number:

R856.5

Zhao JIN,Rushuai WANG,Zhaoyang WANG,zhanbiao HE,Fuming TIAN,Ruijun WANG. Effect of Parnassia palustris Linn extracts on Helicobacter pylori colonization in gastric mucosa and its mechanism[J].Journal of Jilin University(Medicine Edition), 2022, 48(5): 1131-1138.

Figures/Tables 8

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References 20

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Gene	Primer sequence
Cag A	F: 5'-GTCATAATGGCATAGAACCTGAA-3' R: 5'-ATTCCCTAGGGCGTCTAAATAA-3'
Vac A	F: 5'-AAACGACAAGAAAGAGATCAGT-3' R: 5'-CCAGCAAAAGGCCCATCAA-3'
Bab A	F: 5'-AAGCCTATCAAATCCTCCAAACG-3' R: 5'-TGGCGAGCAGTTATTATTCCCT-3'
16S rRNA	F: 5'-CCGCCTACGCGCTCTTTAC-3' R: 5'-CTAACGAATAGCACCGGCTAAC-3'

Group	Bab A	Cag A	Vac A
Hp	15.6±0.61	35.2±2.46	22.4±0.92
Parnassia palustrisLinn	5.93±2.21^*	8.50±3.65^**	8.14±3.37^**

Group	Infection rate	Eradication rate
Hp	86.7	－
Triple	53.3^*	30.8
Quadruple	40.0^*	53.8
Parnassia palustris Linn	46.7^*	46.2
Triple+Parnassia palustris Linn	20.0	76.9^△
Quadruple+Parnassia palustris Linn	13.3	92.3^#
^*P<0.05 vs Hp group; ^△P<0.01 vs triple grouip; ^#P<0.01 vs quadruple group.“－”：No data.

Group	n	IL-2	IL-8	TNF-α
Blank control	12	35.23±3.56	13.33±3.23	6.87±3.26
Normal saline	12	33.52±5.83	17.53±6.25	8.55±4.32
Hp	15	15.71±4.06^*△	34.12±6.22^*△	53.23±9.34^*△
Triple	15	22.12±5.82^#	26.21±5.13^#	40.25±4.12^#
Quadruple	15	25.21±3.43^#	23.25±4.16^#	28.62±5.61^#
Parnassia palustris Linn	15	24.38±4.19^#	20.79±3.32^#	26.32±5.16^##
Triple+Parnassia palustris Linn	15	27.15±5.29^#	19.33±3.87^##	17.77±8.65^##○□
Quadruple+Parnassia palustris Linn	15	29.32±6.22^#	18.26±5.69^##	15.42±4.32^##○□

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Effect of Parnassia palustris Linn extracts on Helicobacter pylori colonization in gastric mucosa and its mechanism

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