Ling Zeng

Comparison of Bush Sophora Root polysaccharide and its sulfate's anti-duck hepatitis A virus activity and mechanism

In order to research the sulfating modification in enhancing the anti-duck hepatitis A virus (DHAV) activity of Bush Sophora Root polysaccharide (BSRPS), sulfated Bush Sophora Root polysaccharide (sBSRPS) was prepared by chlorosulfonic acid-pyridine method. KBr pellets method was applied to analyze their different structures. Anti-DHAV activity was studied by duck embryonic hepatocytes culture in vitro and artificial inoculation method in vivo. Direct immunofluorescence method and Real-time PCR were applied to study the antiviral mechanism of adsorption, replication and release in vitro and the dynamic change of virus content of blood in vivo. The results showed at the most effective content, sBSRPS (7.813 μg/mL) could inhibit both replication and release of DHAV in vitro, BSRPS (500 μg/mL) only inhibit replication. The relative expression of DHAV gene at the 8thh and the mortality rate of sBSRPS group were significantly reduced. These results indicated sBSRPS performed more effectively in anti-DHAV activity than BSRPS.

The anti-DHAV activities of Astragalus polysaccharide and its sulfate compared with those of BSRPS and its sulfate

This paper studied the anti-duck hepatitis A virus (DHAV) activities of Astragalus polysaccharide (APS) and its sulfate (sAPS) compared with those of Bush Sophora Root polysaccharide (BSRPS) and its sulfate (sBSRPS). The antiviral activities of APS and sAPS were measured by MTT and real-time PCR methods, in vitro. In vivo experiment, the mortality rate and the evaluation indexes of hepatic injury, peroxidative injury and immune level were measured. Just like the condition of BSRPS and sBSRPS, the anti-DHAV activities of sAPS were stronger than those of APS, both in vitro and in vivo. It indicated sulfated modification could enhance the antiviral ability of polysaccharide. But unlike the antiviral effects of BSPRS and sBSRPS in vivo, APS and sAPS did not reduce the mortality rates as their abilities of scavenging free radicals and alleviating the hepatic injuries were weaker than those of BSRPS and sBSRPS. And they even did not enhance the immune levels.

Bush Sophora Root polysaccharide and its sulfate can scavenge free radicals resulted from duck virus hepatitis.

In order to study the antioxidant effect of Bush Sophora Root polysaccharide (BSRPS) and its sulfate on anti-duck virus hepatitis (DVH), sulfated Bush Sophora Root polysaccharide (sBSRPS) was prepared by chlorosulfonic acid-pyridine method. Ducklings were fed with BSRPS and sBSRPS after challenged DHAV. Death was monitored, evaluation indexes of peroxidative and hepatic injury at the initial (4th and 8th hour) and later (54th hour) stages were detected. The results showed a fine treatment effect of BSRPS and sBSRPS. Visual hepatic pathological injury severities were less serious after the treatment. At the initial stage, free radical levels in all groups were the same, and BSRPS and sBSRPS reduced the hepatic injury through inhibiting virus replication. At the later stage, mass free radicals were detected in VC group while free radical levels in BSRPS and sBSRPS groups were significantly lower than VC group. The antioxidant effect of BSRPS and sBSRPS might alleviate the hepatic injury.

Anti-duck virus hepatitis mechanisms of Bush Sophora Root polysaccharide and its sulfate verified by intervention experiments

In our previous study, Bush Sophora Root polysaccharide (BSRPS) and its sulfate (sBSRPS) exhibited anti-duck virus hepatitis (DVH) abilities as well as anti-oxidative and immuno-enhancement effects. The aim of this paper was to ulteriorly investigate the exact anti-DVH mechanisms of BSRPS and sBSRPS by intervention experiments. Hinokitiol and FK506 were used as the pro-oxidant and immunosuppressant, respectively. The dynamic deaths, oxidative and immune evaluation indexes and hepatic pathological change scores were detected. When was intervened by hinokitiol, sBSRPS still possessed therapeutic effect while BSPRS was useless. Under the condition of immunosuppression, BSRPS lost a part role in treating DVH; however such a role of sBSRPS completely exhausted. These results suggested both anti-oxidative and immuno-enhancement effects of BSRPS played roles in healing DVH, and the former was more crucial; unlike BSRPS, only immuno-enhancement ability of sBSRPS was imperative for its curative effect on DVH.

Effects of Bush Sophora Root polysaccharide and its sulfate on immuno-enhancing of the therapeutic DVH

Bush Sophora Root polysaccharide (BSRPS) and its sulfate, sulfated Bush Sophora Root polysaccharide (sBSRPS), possess the antiviral activities against duck hepatitis A virus. However their antiviral mechanisms are still not clear. This paper reported their immuno-enhancing roles in the therapeutic effects for duck virus hepatitis (DVH). The effects of BSRPS and sBSRPS on stimulating lymphocyte proliferation were investigated by MTT methods. After that, ducklings were challenged with DHAV and treated with BSRPS and sBSRPS. Meanwhile, the total antibody (Ab), cytokines including interferon gamma (IFN-γ), hepatocyte growth factor (HGF), interleukin (IL)-2, IL-6 and IL-8 were determined by enzyme-linked immuno sorbent assay methods. The results showed that BSRPS owned a fine hepatoprotective effect with stable HGF producing ability. Sulfated modification was able to increase the proliferation rates of B and T lymphocytes and the secretions of total Ab, IFN-γ and IL-2, as comparison with those of BSRPS group. In summary, both of them exhibited immuno-enhancing effects on the therapeutic effects for DVH, and the capacity of sBSRPS was stronger than that of BSRPS.

Treatment effect of a flavonoid prescription on duck virus hepatitis by its hepatoprotective and antioxidative ability

Abstract Context: Duck virus hepatitis (DVH) caused by duck hepatitis A virus type 1 (DHAV-1) is an acute and lethal disease of young ducklings. However, there is still no effective drug to treat DVH. Objective: This study assessed the curative effect on DVH of a flavonoid prescription baicalin-linarin-icariin-notoginsenoside R1 (BLIN) as well as the hepatoprotective and antioxidative effects of BLIN. Materials and methods: MTT method was used to test the anti-DHAV-1 ability of BLIN in vitro. We then treated ducklings by BLIN (3 mg per duckling, once a day for 5 days) to evaluate the in vivo efficacy. To study the hepatoprotective and antioxidative roles of BLIN in its curative effect on DVH, we investigated the hepatic injury evaluation biomarkers and the oxidative stress evaluation indices of the ducklings. Results: On duck embryonic hepatocytes, DHAV-1 inhibitory rate of BLIN at 20 μg/mL was 69.3%. The survival rate of ducklings treated by BLIN was about 35.5%, which was significantly higher than that of virus control (0.0%). After the treatment of BLIN, both the hepatic injury and the oxidative stress of infected ducklings alleviated. At the same time, a significant positive correlation (p < 0.05) existed between the hepatic injury indices and the oxidative stress indices. Conclusions: BLIN showed a significant curative effect on DVH. The antioxidative and hepatoprotective effects of BLIN made great contributions to the treatment of DVH. Furthermore, BLIN is expected to be exploited as a new drug for the clinical treatment of DVH.

Assessment of a Flavone-Polysaccharide Based Prescription for Treating Duck Virus Hepatitis.

Because polysaccharide and flavone ingredients display good antiviral activity, we developed a flavone/polysaccharide-containing prescription that would be effective against duck viral hepatitis (DVH) and investigated its hepatoprotective effects. Flavones were derived from Hypericum japonicum (HJF) (entire herb of Hypericum japonicum Thunb) and Salvia plebeia (SPF) (entire herb of Salvia plebeia R. Br.), and polysaccharides were derived from Radix Rehmanniae Recens (RRRP) (dried root of Rehmannia glutinosa Libosch). This prescription combination was based on the theory of syndrome differentiation and treatment in traditional Chinese veterinary medicine. In vitro and in vivo experiments were conducted using the three single ingredients compared to the combined HRS prescription to determine their anti-duck hepatitis A viral (anti-DHAV) activity. The results showed that all experimental conditions displayed anti-DHAV activity, but the HRS prescription presented the best effect. To further investigate the hepatoprotective effect of the HRS prescription on DHAV-induced hepatic injury, we tested the mortality rate, the hepatic pathological severity score, plasma biochemical indexes of hepatic function, blood DHAV gene expression levels and peroxidation damage evaluation indexes and then analyzed correlations among these indexes. The results demonstrated that the HRS prescription significantly decreased the mortality rate, reduced the severity of hepatic injury, decreased the hepatic pathological severity score, depressed blood DHAV gene expression levels, and returned the indexes of hepatic function and peroxidation almost to a normal level. These results indicate that the HRS prescription confers an outstanding hepatoprotective effect, and we expect that it will be developed into a new candidate anti-DHAV drug.

Anti-DHAV-1 reproduction and immuno-regulatory effects of a flavonoid prescription on duck virus hepatitis

Abstract Context: The flavonoid prescription baicalin-linarin-icariin-notoginsenoside R1 (BLIN) has a curative effect on duck virus hepatitis (DVH) caused by duck hepatitis A virus type 1 (DHAV-1). However, the mechanism of this curative effect is not understood. Objective: This study investigates the mechanism of the curative effect of BLIN on DVH caused by DHAV-1. We analyzed the anti-DHAV-1 reproduction mechanism and immuno-regulatory effect of BLIN. Materials and methods: The anti-DHAV-1 reproduction effects of BLIN at 20, 10, 5 and 2.5 μg/mL in vitro, as well as the influence of BLIN at 20 μg/mL on DHAV-1 adsorption, replication and release were tested using the qRT-PCR method. The promotion abilities of BLIN at 20, 10, 5 and 2.5 μg/mL on T- and B-lymphocyte proliferation were investigated by the MTT method. IL-2 and IFN-γ levels and total anti-DHAV-1 antibody secretion after treatment with DHAV-1 for 4, 8 and 54 h were determined by ELISA. Results: BLIN showed a dose-dependent DHAV-1 reproduction inhibitory effect. The inhibitory effect was highest at 20 μg/mL, where DHAV-1 adsorption and release were significantly lower. Meanwhile, BLIN at 5 μg/mL significantly increased T and B lymphocyte proliferation. BLIN stimulated total anti-DHAV-1 antibody secretion in ducklings at the dosage of 4 mg per duckling, but did not stimulate IL-2 and IFN-γ secretion significantly. Conclusions: BLIN inhibits DHAV-1 reproduction by suppressing its adsorption and release. Additionally, BLIN promoted the duckling antiviral response.

Effects of Bush Sophora Root polysaccharide and its sulfate on DHAV-1 replication

Duck hepatitis A virus type 1 (DHAV-1) is a mortal virus to ducklings under three weeks old. Bush sophora root polysaccharide (BSRPS) and its sulfate, sulfated BSRPS (sBSRPS), inhibited the replication of DHAV-1. However the detailed mechanisms were still unknown. The protein translation and RNA synthesis are two most important steps of the replication. Additionally, DHAV-1 3D protein, cellular Hsp70 protein, and DHAV-1 IRES were the important regulatory factors. Therefore, the influence of BSRPS and sBSRPS on DHAV-1 protein translation and RNA synthesis were studied. RT-qPCR, western blot, and dual-luciferase reporter system were used to study the inhibition mechanisms of BSRPS and sBSRPS on DHAV-1. The results showed both BSRPS and sBSRPS significantly inhibited the DHAV-1 protein translation and RNA synthesis, and the effect of sBSRPS was stronger. Furthermore, they dropped the protein translation via suppressing DHAV-1 IRES activity and dropped the DHAV-1 synthesis via suppressing cellular Hsp70 expression.

RAW REHMANNIA RADIX POLYSACCHARIDE CAN EFFECTIVELY RELEASE PEROXIDATIVE INJURY INDUCED BY DUCK HEPATITIS A VIRUS

Background: Duck viral hepatitis (DVH), caused by duck hepatitis A virus (DHAV), is a fatal contagious infectious disease which spreads rapidly with high morbidity and high mortality, and there is no effective clinical drug against DVH. Materials and Methods: Raw Rehmannia Radix Polysaccharide (RRRP), Lycii Fructus polysaccharides and Astragalus Radix polysaccharides were experimented in vitro and in vivo. Mortality rate, livers change, liver lesion scoring, peroxidative injury evaluation indexes in vitro and in vivo, and hepatic injury evaluation indexes of optimal one were detected and observed in this experiment. Results: RRRP could reduce mortality with the protection rate about 20.0% compared with that of the viral control (VC) group, finding that RRRP was the most effective against DHAV. The average liver scoring of the VC, blank control (BC), RRRP groups were 3.5, 0, 2.1. Significant difference (P<0.05) appeared between any two groups, demonstrating that it can alleviate liver pathological change. RRRP could make the hepatic injury evaluation indexes similar to BC group while the levels of the VC group were higher than other two groups in general. The levels of SOD, GSH-Px, CAT of RRRP group showed significant higher than that of VC group while the levels of NOS and MDA showed the opposite tendency, thus, RRRP could release peroxidative injury. Conclusion: RRRP was the most effective against duck hepatitis A virus (DHAV). RRRP could reduce mortality, alleviate liver pathological change, down-regulate liver lesion score, release peroxidative injury and hepatic injury. The antiviral and peroxidative injury releasing activity of RRRP for DHAV provided a platform to test novel drug strategies for hepatitis A virus in human beings.