Cerebral ischemia is a serious neurological disorder that can lead to high morbidity and mortality. Baicalin is a naturally bioactive flavonoid derived from Scutellaria baicalensis Georgi, which has neuroprotective activity. Baicalin exerts a neuroprotective effect against hypoxic ischemic injury. In this study, we investigated whether baicalin regulates specific proteins in the cerebral cortex of ischemic stroke animals. Middle cerebral artery occlusion (MCAO) surgery was performed to induce ischemic brain injury, and baicalin (30 mg/kg) or vehicle was injected into the abdominal cavity before MCAO surgery. Neurological behavior tests were performed 24 h after MCAO surgery and proteomics approach was performed using proteins extracted from cortical tissue. Two-dimensional electrophoresis analysis and MALDI-TOF were performed to identify the regulated protein by baicalin. MCAO damage caused severe behavioral disorders, but baicalin treatment improved these behavioral deficits. Baicalin also induced changes in the expression of various proteins in the cerebral cortex of MCAO animals. Proteins changed by baicalin administration are as follow: adenosylhomocysteinase, isocitrate dehydrogenase [NAD] subunit alpha, apolipoprotein A-I, Rab GDP dissociation inhibitor beta, eukaryotic initiation factor 4A, and mu-crystallin. These proteins were involved in metabolism and protein synthesis. The results of this study demonstrated the neuroprotective effects of baicalin by improving behavioral disorders caused by MCAO damage. The results also showed that baicalin regulates the expression of a variety of proteins involved in neuroprotective functions. Therefore, our findings provide evidence that baicalin plays a neuroprotective role in stroke animal models by regulating specific proteins.

Periodontal disease is an inflammatory disease that affects the destruction of the bone supporting the tooth and connective tissues surrounding it. Periodontal ligament fibroblasts (PDLFs) induce overexpression of matrix metalloproteinase (MMP) involved in periodontal diseaseʼs inflammatory destruction. Osteoclasts take part in physiological bone remodeling, but they are also involved in bone destruction in many kinds of bone diseases, including osteoporosis and periodontal disease. This study examined the effect of baicalin on proteolytic enzymesʼ production and secretion of inflammatory cytokines in PDLFs and RAW 264.7 cells under the lipopolysaccharide (LPS)-induced inflammatory conditions. Baicalin inhibited the expression of the protein, MMP-1 and MMP-2, without affecting PDLFs’ cell viability, suggesting its possibility because of the inhibition of phosphorylation activation of mitogen-activated protein kinase’s p38, and the signal transduction process of nuclear factor κB (NFκB)-related protein. Also, baicalin reduced the expression of MMP-8 and MMP-9 in RAW 264.7 cells. This reduction is thought to be due to the inhibition of the signal transduction process of NFκB-related proteins affected by inhibiting p65RelA phosphorylation. Also, baicalin inhibited the secretion of nitric oxide and interleukin-6 induced by LPS in RAW 264.7 cells. These results suggest that baicalin inhibits connective tissue destruction in periodontal disease. The inhibition of periodontal tissue destruction may be a therapeutic strategy for treating inflammatory periodontal-diseased patients.

Baicalin is a flavonoid compound with many advantages, including anti-inflammatory agents and antioxidants. Lipopolysaccharide (LPS) is an endotoxin that induces neuronal damage through inflammatory response and oxidative stress reaction. This study was investigated the protective effects of baicalin on the oxidative stress and histopathological changes caused by LPS in hippocampus. Adult mice were divided into four groups; vehicle-treated, baicalin-treated, LPS-treated, and LPS and baicalin co-treated animals. Baicalin (10 mg/kg/day) and/or LPS (250 μg/kg/day) were intraperitoneally administered for seven consecutive days, and body weight was measured. Reactive oxygen species (ROS) level and lipid peroxidation level in the hippocampus were examined. Histopathological study was performed using hematoxylin and eosin staining manuals. LPS treatment decreased body weight and increased ROS and oxidative stress in the hippocampus. However, co-treatment with baicalin alleviated these changes caused by LPS. Severe histopathological changes were observed in the hippocampus of LPS-treated animals. Baicalin co-treatment attenuated the changes and preserved neuronal cells from LPS damage. These results showed that baicalin suppresses LPS-induced neuronal damage by alleviating oxidative stress in the hippocampus. Thus, this study demonstrated that baicalin exerts protective effects against LPS-induced oxidative stress in hippocampus.

본 연구는 건강기능식품 중 baicalin, eleutheroside E, ligustilide를 효과적으로 분석할 수 있는 방법을 확립하기 위하여 수행되었다. 이에 LC-MS/MS를 이용하여 신속하고 효율적으로 동시분석할 수 있는 시험법을 확립하였으며, 확립된 시험법에 대해 특이성, 검출한계, 정량한계, 정확도, 정밀도에 대한 검증을 통하여 유효성을 확인하고자 하였다. 표준용액을 이용하여 검량선을 작성한 결과 r2> 0.99 이상의 직선성을 확인하였고, baicalin, eleutheroside E, ligustilide에 대한 정량한계는 각각 39.3 μg/L, 106.7 μg/L, 76.1 μg/L이었으며, 검출한계는 각각 13.0 μg/L, 35.2 μg/L, 25.1 μg/L이었다. 또한 평균 회수율은 각 성분에 대해 108.0~109.9%, 99.8~101.3%, 91.4~97.2%로 나타났으며, 반 복정밀도는 상대표준편차 5%이하, 실험실간 재현성은 9% 이하로 나타나 정확성, 재현성이 우수하였으며 이는 AOAC 가이드라인19)에서 제시한 기준에 모두 적합한 수준이었다. 따라서 개발된 분석법은 향후 건강기능식품 중 baicalin, eleutheroside E, ligustilide를 동시분석하는데 효과적으로 활용할 수 있을 것으로 판단된다.

This study examined the anti-osteoclastogenic effects of baicalin on receptor activator of NF-kB ligand (RANKL)- induced RAW264.7 cells. Baicalin is a flavonoid that is produced by Scutellaria baicalensis and is known to have multiple biological properties, including antibacterial, anti- inflammatory and analgesic effects. The effects of baicalin on osteoclasts were examined by measuring 1) cell via- bility; 2) the formation of tartrate-resistant acid phosphatase (TRAP) (+) multinucleated cells; 3) RANK/RANKL signa- ling pathways and 4) mRNA levels of osteoclast-associated genes. Baicalin inhibited the formation of RANKL-stimu- lated TRAP (+) multinucleated cells and also suppressed the RANKL-stimulated activation of p-38, ERK, cSrc and AKT signaling. Baicalin also inhibited the RANKL-stimu- lated degradation of IĸB in RAW264.7 cells. In addition, the RANKL-stimulated induction of NFATc1 transcription factors was found to be abrogated by this flavonoid. Baica- lin was further found to decrease the mRNA expression of osteoclast-associated genes, including carbonic anhydrase II, TRAP and cathepsin K in the RAW264.7 cells. Our data thus demonstrate that baicalin inhibits osteoclastogenesis by inhibiting the RANKL-induced activation of signaling molecules and transcription factors in osteoclast precursors.

Baicalin is a flavonoid purified from the medicinal plant Scutellaria baicalensis. It has been reported that baicalin exhibits antibacterial, anti-inflammatory and analgesic effects. The present study was undertaken to determine the underlying cellular mechanisms of baicalin action in preosteoclasts. The effects of this flavonoid on preosteoclasts were determined by measuring osteoclast generation and osteoclast activity in macrophage-colony stimulating factor (M-CSF)-dependent bone marrow cells (MDBMCs) and in co-cultures of MDBMCs and osteoblasts. Osteoclast generation was assayed by measuring the number of tartrateresistant acid phosphatase (TRAP) (+) multinucleated cells after culture. Osteoclast activity was assayed by measuring the area of the resorption pit after culture. We found that osteoclast generation was induced by M-CSF and receptor activator of NF-kB ligand (RANKL), and by the 1.25-dihydroxycholecalciferol in our cultures. Baicalin decreased both osteoclast generation and activity in MDBM cultures and co-cultures indicating that it may inhibit bone resorption.

Scutellaria baicalensis Georgi (SJ) is a perennial plant and its root has been used in oriental traditional medicine for treatment of fever, inflammation, diarrhea and anticancer effect, etc. In this study, plant tissue culture system for SJ was developed. Stem piece of younger plant was optimum explant for callus induction and growth on MS medium supplemented with NAA 1.0 ㎎/L plus BA 0.5 ㎎/L. Plantlet regeneration through callus culture was well on MS medium containing NAA 1.0 mg/L. SJ has been known biologically active substances such as baicalin, baiclein, and wogonin. This study was carried out to examine the effect of plant growth regulators for production of baicalin, baicalein, and wogonin through callus culture. The HPLC pattern of callus extract was identical to that of standard solution, it shows that the callus produced by tissue culture has the same flavonoids composition of SJ. Baicalin, baicalein, and wogonin production was 471.5~52.8 ㎍/g, 137.6~4.0 ㎍/g, and 16.6~1.3 ㎍/g, respectively, on MS media with nine different plant growth regulator combinations. This may indicate that plant tissue culture of SJ possible to produce the biologically active substances effectively.