Zhihui Jiang

Echinacoside and Cistanche tubulosa (Schenk) R. wight ameliorate bisphenol A-induced testicular and sperm damage in rats through gonad axis regulated steroidogenic enzymes

ETHNOPHARMACOLOGICAL RELEVANCE Male infertility has been increasing over the last decades and being a pressing health problem nowadays. Cistanche tubulosa (CT) is a traditional Chinese medicine used to boost male sexual function. Echinacoside (ECH) is one of the major compounds exist in CT and might be a potential agent to protect testis and sperm injury. AIM OF THE STUDY To investigate the mechanisms behind the possible protective effects of CT and ECH against testicular and sperm toxicity. MATERIALS AND METHODS CT was identified by 5.8s gene sequencing. The major compositions (echinacoside and acteoside) of CT were quantified by HPLC method. The adult male Sprague-Dawley rats were exposed to BPA, CT or ECH for 42 consecutive days. The sperm parameters were observed by dark-field microscope; serum hormone levels (FSH, LH and testosterone) were tested by radio immunosorbent; LDH-x activity were evaluated using commercial kits; the expressions of the key steroidogenic enzymes were evaluated by qRT-PCR, heat map, immunofluorescence and western blot. RESULTS The CT and ECH treatments against BPA-induced testicular and sperm toxicity showed that CT and ECH have reversed BPA-induced abnormality in sperm characteristics, testicular structure and normalized serum testosterone. This was concomitant with the increased expression of LDH-x as well as the key steroidogenic enzymes including StAR, CYP11A1, 3β-HSD, 17β-HSD and CYP17A1, suggesting that CT and ECH enhanced testosterone biosynthesis. CONCLUSIONS CT and ECH attenuated poor sperm quality and testicular toxicity in rats through up-regulation steroidogenesis enzymes and ECH is the active compound of CT as a potential natural reproductive agent.

Hepatoprotective Effect of Kaempferol Against Alcoholic Liver Injury in Mice

Kaempferol is a biologically active component present in various plants. The hepatoprotective effect of kaempferol in drug-induced liver injury has been proven, while its effect against alcoholic liver injury (ALI) remains unclear. Hence, the present study aimed to evaluate the effect of kaempferol against ALI in mice. The experimental ALI mice model was developed and the mice were treated with different doses of kaempferol for 4 weeks. The liver functions were observed by monitoring the following parameters: Aspartate aminotransferase (AST/GOT) and alanine aminotransferase (ALT/GPT) levels in serum; histopathological studies of liver tissue; oxidative stress by hydrogen peroxide (H2O2), superoxide dismutase (SOD) and glutathione (GSH); the lipid peroxidation status by malondialdehyde (MDA) and lipid accumulation by triglyceride (TG) level in serum; and the expression levels and activities of a key microsomal enzyme cytochrome 2E1 (CYP2E1), by both in vitro and in vivo methods. The ALI mice (untreated) showed clear symptoms of liver injury, such as significantly increased levels of oxidative stress, lipid peroxidation and excessive CYP2E1 expression and activity. The mice treated with different kaempferol dosages exhibited a significant decrease in the oxidative stress as well as lipid peroxidation, and increased anti-oxidative defense activity. The kaempferol treatment has significantly reduced the expression level and activity of hepatic CYP2E1, thus indicating that kaempferol could down regulate CYP2E1. These findings show the hepatoprotective properties of kaempferol against alcohol-induced liver injury by attenuating the activity and expression of CYP2E1 and by enhancing the protective role of anti-oxidative defense system.

Protective effect of hyperoside against acetaminophen (APAP) induced liver injury through enhancement of APAP clearance.

Acetaminphen (APAP) overdose leads to severe hepatotoxicity. Apocynum venetum L. (A. venetum) possess potent hepatoprotective effect. Hyperoside is one of the major compounds exist in Apocynum venetum L. and might be a potential agent to protect against APAP-induce liver injury. In this study, we investigated the effect of hyperoside on APAP hepatotoxicity in mice. Mice were treated intragastrically with hyperoside (10, 50 or 100 mg/kg) for 3 days before APAP (300 mg/kg) injection. APAP alone caused severe liver injury characterized by significantly increased serum aminotransferase levels, hepatic malondialdehyde (MDA) and 3-nitrotyrosine (3-NT) formation, as well as liver superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione (GSH) depletions. Hyperoside significantly attenuated APAP-induced liver damages in a dose dependent manner, and 100 mg/kg was the most effective dose. Further study confirmed that hyperoside was able to increase activities and mRNA expressions of uridine diphoshate glucuronosyltransferases (UGTs) and sulfotransferases (SULTs), as well as to inhibit CYP2E1 activities, and thereby suppressed toxic intermediate formation and promoted APAP hepatic detoxification. Nrf-2 activation might be involved in hyperoside induced up-regulation of phase II enzymes. Collectively, our data provide evidence that hyperoside protected the liver against APAP induced injury mainly by accelerating APAP harmless metabolism, implying that hyperoside can be considered as a potential natural hepatoprotective agent.

3,4-Dihydroxyphenylacetic acid, a microbiota-derived metabolite of quercetin, attenuates acetaminophen (APAP)-induced liver injury through activation of Nrf-2.

Abstract 1. Acetaminophen (APAP) overdose leads to severe hepatotoxicity. 3,4-dihydroxyphenylacetic acid (DOPAC) is a scarcely studied microbiota-derived metabolite of quercetin. The aim of this study was to determine the protective effect of DOPAC against APAP-induced liver injury. 2. Mice were treated intragastrically with DOPAC (10, 20 or 50 mg/kg) for 3 days before APAP (300 mg/kg) injection. APAP alone caused increase in serum aminotransferase levels and changes in hepatic histopathology. APAP also promoted oxidative stress by increasing lipid peroxidation and decreasing anti-oxidant enzyme activities. These events led to hepatocellular necrosis and reduced liver function. DOPAC increased nuclear factor erythroid 2-related factor 2 (Nrf-2) translocation to the nucleus and enhanced the expression of phase II enzymes and anti-oxidant enzymes, and thereby reduced APAP hepatotoxicity and enhanced anti-oxidant ability. 3. Our data provide evidence that DOPAC protected the liver against APAP-induced injury, which is involved in Nrf-2 activation, implying that DOPAC can be considered as a potential natural hepatoprotective agent.

Purple potato (Solanum tuberosum L.) anthocyanins attenuate alcohol-induced hepatic injury by enhancing antioxidant defense

Alcoholic liver disease (ALD) is a serious and challenging health issue. In the past decade, natural components possessing hepatoprotective properties have gained more attention for ALD intervention. In this study, the phytochemical components of anthocyanins from purple potato were assessed using UPLC-MS/MS, and the hepatoprotective effects of purple potato anthocyanins (PPAs) were investigated in the ALD mouse model. Serum and liver biochemical parameters were determined, along with histopathological changes in liver tissue. In addition, the major contributors to alcohol-induced oxidative stress were assessed. The results indicated that the levels of aspartate transaminase and alanine transaminase were lower in the serum of the PPA-treated group than the alcohol-treated group. PPAs significantly inhibited the reduction of total cholesterol and triglycerides. Higher levels of superoxide dismutase and reduced glutathione enzymes as well as a reduction in the formation of malondialdehyde occurred in mice fed with PPAs. In addition, PPAs protected against increased alcohol-induced levels and activity of cytochrome P450 2E1 (CYP2E1), which demonstrates the effects of PPAs against alcohol-induced oxidative stress and liver injury. This study suggests that PPAs could be an effective therapeutic agent in alcohol-induced liver injuries by inhibiting CYP2E1 expression and thereby strengthening antioxidant defenses.

Anthocyanins attenuate alcohol-induced hepatic injury by inhibiting pro-inflammation signalling

We assessed phytochemical components of anthocyanins from purple sweet potato (PSP) and purple potato (PP) with UPLC-MS/MS, and investigated their inhibitory effect on inflammatory response in alcoholic liver disease (ALD). Results showed that serum AST and ALT levels in PP anthocyanins (PPAs) and PSP anthocyanins (PSPAs) treatments were lower than those of alcohol-treated group. PPAs and PSPAs could inhibit mRNA expressions of inflammatory factors (TNF-α, VCAM-1, IFN-γ and CXCL-1). The mRNA levels of NF-κB, STAT, and TLR in PPAs and PSPAs treatment groups were lower than in alcohol treatment group. Our results indicate that PP and PSP are good source of anti-inflammatory anthocyanins to prevent ALD.

Apocynum venetum Attenuates Acetaminophen-Induced Liver Injury in Mice

Apocynum venetum L. (A. venetum) has long been used in oriental folk medicine for the treatment of some liver diseases; however, the underlying mechanisms remain to be fully elucidated. Acetaminophen (APAP) is a widely used analgesic drug that can cause acute liver injury in overdose situations. In this study, we investigated the potential protective effect of A. venetum leaf extract (ALE) against APAP-induced hepatotoxicity. Mice were intragastrically administered with ALE once daily for 3 consecutive days prior to receiving a single intraperitoneal injection of APAP. The APAP group showed severe liver injury characterized by the noticeable fluctuations in the following parameters: serum aminotransferases; hepatic malondialdehyde (MDA), 3-nitrotyrosine (3-NT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione (GSH). These liver damages induced by APAP were significantly attenuated by ALE pretreatments. A collective analysis of histopathological examination, DNA laddering and western blot for caspase-3 and cytochrome c indicated that the ALE is also capable of preventing APAP-induced hepatocyte death. Hyperoside, isoquercitrin and their derivatives have been identified as the major components of ALE using HPLC-MS/MS. Taken together, the A. venetum possesses hepatoprotective effects partially due to its anti-oxidant action.

Protective effect of Cordyceps militaris extract against bisphenol A induced reproductive damage

ABSTRACT This study aimed to investigate the protective effects of Cordyceps militaris (C. militaris) against reproductive damage induced by bisphenol A (BPA). Rats were administrated 200 mg/kg BPA for 4 weeks and treated with C. militaris (200, 400, and 800 mg/kg body weight/day). By the end of the fourth week, the level of oxidative damage, sperm parameters, hormone levels, and histopathological changes were examined. In the group that only received BPA, there was a significant decrease in body weight compared with the normal control (NC) group. C. militaris significantly alleviated the BPA-induced reproductive damage by increasing testicular superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), and glutathione (GSH); as well as by reducing serum malondialdehyde (MDA). C. militaris not only obviously enhanced the levels of serum LH and T, but it also improved the sperm count and motility compared to the BPA-treated group. These results suggest that C. militaris could be used as a potential natural substance for preventing BPA induced reproductive damage. Abbreviations BPA: bisphenol A; SOD: superoxide dismutase; GSH: glutathione; GSH-PX: glutathione peroxidase; MDA: malondialdehyde; ROS: reactive oxygen species; T: testosterone; LH: luteinizing hormone; FSH: follicle-stimulating hormone; UPLC: ultra performance liquid chromatography; RIA: radioimmunoassay; qRT-PCR: quantitative real time PCR; NC: normal control group; BPA: 200 mg/kg BPA administered group; H: 800 mg/kg C. militaris extract administered group; LB, MB, and HB: 200 mg/kg BPA + 200 mg/kg, 400 mg/kg, and 800 mg/kg C. militaris administered group, respectively; VeB: 200 mg/kg BPA + 300 mg/kg Vitamin E administered group; Star: steroidogenic acute regulatory protein; 3β-HSD: 3beta-hydroxyl-delta-5-steroid dehydrogenase; CYP11A1: cytochrome P 450 family 11 subfamily A member 1; CYP17A1: cytochrome P 450 family 17 subfamily A member 1

Protective effect of wild Corni fructus methanolic extract against acute alcoholic liver injury in mice

ABSTRACT Background: In Chinese folk medicine, Corni fructus (C. fructus) has traditionally been used to improve liver function, although the mechanism underlying its activity remains unclear. The aim of the present study was to evaluate the protective effects of wild C. fructus methanolic extract against acute alcoholic liver injury. Methods: Alcohol was administered to mice for three consecutive days, either alone or in combination with C. fructus methanolic extract (50, 100, or 200 mg/kg body weight/d). Serum and liver tissue were collected from the animals and subjected to biochemical and histopathological analyses. Results: C. fructus significantly alleviated alcohol-induced liver injury by reducing serum alanine aminotransferase, aspartate aminotransferase, and thiobarbituric acid reactive species, inhibiting hydroxyl radicals (•OH), and increasing total superoxide dismutase, glutathione peroxidase, and glutathione in the liver (P < 0.05). In addition, the C. fructus treatment inhibited the expression and activity of cytochrome P450 2E1 (P < 0.05). Conclusions: C. fructus could be a promising natural substance for ameliorating acute alcohol-induced oxidative stress and hepatic injury. GRAPHICAL ABSTRACT

Echinacoside Increases Sperm Quantity in Rats by Targeting the Hypothalamic Androgen Receptor

Male infertility is a major health issue with an estimated prevalence of 4.2% of male infertility worldwide. Our early work demonstrated that Cistanche extracts protect against sperm damage in mice and that echinacoside (ECH) is one of the major active components. Here we report an essential role for ECH, a natural product that reverses or protects against oligoasthenospermia in rats. ECH was assayed by HPLC, the quantity and quality of sperm was evaluated and hormone levels were determined by radioimmunosorbent assay. ECH reduced levels of androgen receptor (AR) and key steroidogenic-related genes as determined by Western blot and qPCR analysis. The interaction between ECH and AR were evaluated by indirect ELISA and molecular docking. The results show that ECH combined with hypothalamic AR in the pocket of Met-894 and Val-713 to inhibit transfer of AR from the cytoplasm to nuclei in the hypothalamus. While negative feedback of sex hormone regulation was inhibited, positive feedback was stimulated to increase the secretion of luteinizing hormone and testosterone subsequently enhancing the quantity of sperm. Taken together, these data demonstrate that ECH blocks AR activity in the hypothalamus to increase the quantity of sperm and protect against oligoasthenospermia in rats.