Study On The Protective Effect Of Cistanche Phenylethanol Glycosides On Rat Sperm Oxidative Injury in Vitro

Li Gang, Zhu Wenbin, Niu Fei, Zhang Hongli

Inner Mongolia Medical College·School of Pharmacy, Hohhot, Inner Mongolia

Contact: joanna.jia@wecistanche.com / WhatsApp: 008618081934791

Abstract

Objective To observe the interventional effects of Cistanche phenyl ethanol glycosides on sperm membrane structure and oxidative damage in rats, and to explore the mechanism of Cistanche phenyl ethanol glycosides in the treatment of male infertility with asthenospermia. Methods Sperm suspension was prepared from rat sperm and divided into normal group, drug blank group, model group, positive drug control group (vitamin C group) and Cistanche cistanche phenyl ethanol glycoside extract small-medium, and high-dose groups, using FeSO4/ The H2O2 system produces reactive oxygen species (ROS). In an aerobic environment, different doses (0.025, 0.25, 0.5 g/ml crude drug amount) of Cistanche phenoxyethanol glycosides are incubated with sperm suspension to detect the lipid peroxidation damage of the sperm membrane The degree of sperm membrane function was evaluated by the sperm hypotonic swelling test, and compared with the known antioxidant vitamin C. Results： The small, medium, and high-dose Cistanche phenyl ethanol glycoside groups (0.025, 0.25, 0.5 g·ml-1) under the same conditions can increase the SOD activity of the sperm suspension, reduce the MDA content, and have a certain degree of protection for the sperm membrane function effect. Among them, the Cistanche phenoxyethanol glycoside group of 0.25, 0.5 g·ml-1 was significantly better than the vitamin C group (P<0.01 or P<0.001). Conclusion Cistanche phenethyl alcohol glycosides have obvious intervention effects on the lipid peroxidation damage of rat sperm membranes and have obvious protective effects on the structure and function of sperm membranes.

Keywords: Cistanche; Phenylethanol glycoside; Sperm; Lipid peroxidation

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Cistanche, as a precious Chinese medicinal material, has been studied more in recent years. The active ingredients of Cistanche cistanche mainly include phenylethanoid glycosides (PhGs), cistanche polysaccharides, betaine, flavonoids, and other substances, among which PhGs are one of the most important active ingredients in cistanche. PhGs are a class of compounds containing hydroxy, methoxy substituted phenethyl and hydroxy, methoxy substituted cinnamoyl, usually β-glucose as the parent nucleus containing ester bonds, and oxo-glycosidic bonds natural glycosides, widely present in Gemini Leaf plant. Phenyl ethanol glycosides are mainly composed of caffeic acid, phenethyl aglycones, and sugars, including echinacoside (Echi-nacoside), ergosteroside (Acteoside), isoergoside (Isoacteoside), 2-acetyl ergoster Glycoside (2'-acetylacteoside), and so on. Many research results show that phenyl ethanol glycosides have antibacterial, anti-inflammatory, anti-viral, anti-tumor, anti-oxidant, immune regulation, memory, and sexual function enhancement effects. Because PhGs molecules contain multiple phenolic hydroxyl groups, they can bind free radicals in the body, avoid their accumulation in the body, protect the body from excessive oxidative damage, and are the main components of anti-oxidation and anti-aging [1].

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Although Cistanche has repeatedly appeared in the Chinese medicine compatibility in the treatment of male deficiency and weak essence in the form of compound [2], the research on the mechanism of its effect is still incomplete. In addition to the androgen and gonadotropin-like effects it can exhibit to improve sperm count and vitality [3], it is a new attempt to study its antioxidant effect in the treatment of weak and oligospermia diseases. In this paper, with the topic of the protective effect of Cistanche cistanche extract on the oxidative damage of rat sperm outer membrane function, in vitro application of FeSO4 /H2O2 system to establish oxidative stress damage [4], to explore the role of Cistanche's antioxidant effect in the treatment of such diseases.

Discussion

ROS includes oxygen ions, free radicals, and superoxide anions, which are produced by sperm and sperm granulocytes and cause infertility through two main mechanisms. First, it destroys the sperm cell membrane, reducing sperm motility and the ability to bind to the egg; second, ROS can change the sperm DNA, causing defective paternal DNA to act on the pregnancy. We know that the sperm membrane is rich in polyunsaturated fatty acids, which are extremely unstable under aerobic conditions and is prone to lipid peroxidation (LPO) of sperm membrane, which changes the liquidity, fluidity, and permeability of the membrane. , Causing damage to the sperm membrane and even cell death. As one of the final products of LPO reaction, MDA content can reflect the damage of sperm membrane LPO to a certain extent [9]. SOD is the main enzyme system possessed by sperm to defend against ROS. Hypotonic swelling test: an alternative test method recommended by WHO to check sperm viability. Some scholars believe that the hypotonic sperm swelling test can be used as one of the indicators of sperm membrane function.

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In this paper, the FeSO4/H2O2 system was used to generate ROS, which caused the oxidative stress damage model. The protective effect of Cistanche cistanche extract on the structure and function of rat sperm membrane was studied. The results showed that under the action of ROS, the structure and function of the rat sperm membrane were significantly damaged, the activity of sperm suspension T-SOD was significantly reduced, the content of MDA was significantly increased, and the sperm swelling rate (total swelling rate, g-sperm swelling rate) was significantly reduced. , Sperm survival rate decreased significantly, and the difference was extremely significant compared with the normal group (P<0.001). The small, medium and high-dose groups of Cistanche extract (0.025, 0.25, 0.5 g·ml-1) under the same conditions can increase the SOD activity of the sperm suspension, reduce the MDA content, and cause oxidative damage to the sperm membrane caused by ROS. It has different degrees of intervention and has a certain protective effect on the function of the sperm membrane. The effect of Cistanche cistanche extract is dose-dependent, and the protective effect on sperm membrane function is the most significant at high dose (0.5 g·ml-1). Cistanche has anti-oxygen free radical activity and has a good scavenging effect on hydroxyl free radicals and superoxide anion free radicals.

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The results of this experiment show that an appropriate amount of Cistanche extract can significantly increase the SOD activity of the sperm suspension, reduce the MDA content, inhibit the lipid peroxidation of the sperm membrane, exert an antioxidant effect, and have a significant protective effect on the structure and function of the sperm membrane. This may be another new pharmacological mechanism of Cistanche in the treatment of male infertility with asthenospermia in addition to the existing androgen and gonadotropin-like effects.

References

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