Alcohol extract of root and root bark of Toddalia asiatica alleviates CIA in rats through anti-inflammatory and proapoptotic effects / 中国中药杂志

This study aims to investigate the therapeutic effect of alcohol extract of root and root bark of Toddalia asiatica(TAAE) on collagen-induced arthritis(CIA) in rats through phosphatidylinoinosidine-3 kinase/protein kinase B(PI3K/Akt) signaling pathway. To be specific, CIA was induced in rats, and then the rats were treated(oral, daily) with TAAE and Tripterygium Glycoside Tablets(TGT), respectively. The swelling degree of the hind leg joints was scored weekly. After 35 days of administration, the histopathological changes were observed based on hematoxylin and eosin(HE) staining. Enzyme-linked immunosorbent assay(ELISA) was employed to detect the levels of cytokines [tumor necrosis factor-α(TNF-α), interleukin(IL)-6)]. Terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL) staining was performed to detect the apoptosis of synoviocytes in rats. Western blot was used to detect the expression levels of apoptosis-related proteins B-cell lymphoma 2(Bcl-2)-associated X(Bax), Bcl-2, and caspase-3 and pathway-related proteins phosphoinositide 3-kinase(PI3K), phosphorylated(p)-PI3K, protein kinase B(Akt), and p-Akt. RT-qPCR was conducted to examine the mRNA levels of Bax, Bcl-2, caspase-3, TNF-α, IL-6, and IL-1β and pathway-related proteins PI3K, p-PI3K, Akt, and p-Akt. TAAE can alleviate the joint swelling in CIA rats, reduce serum levels of inflammatory cytokines, improve synovial histopathological changes, promote apoptosis of synoviocytes, and inhibit synovial inflammation. In addition, RT-qPCR and Western blot results showed that TAAE up-regulated the level of Bax, down-regulated the level of Bcl-2, and activated caspase-3 to promote apoptosis in synoviocytes. TAAE effectively down-regulated the protein levels of p-PI3K and p-Akt. In this study, TAAE shows therapeutic effect on CIA in rats and reduces the inflammation. The mechanism is that it suppresses PI3K/Akt signaling pathway and promotes synoviocyte apoptosis. Overall, this study provides a new clue for the research on the anti-inflammatory mechanism of TAAE and lays a theoretical basis for the better clinical application of TAAE in the treatment of inflammatory and autoimmune diseases.

Effect of Alcohol Extract of Oroxylum indicum on Reducing Uric Acid and Protecting Kidney in Hyperuricemia Mice / 中国实验方剂学杂志

ObjectiveTo investigate the effect of alcohol extract of Oroxylum indicum （MHD-80） on reducing uric acid （UA） and protecting the kidney in the hyperuricemia （HUA） model in vivo. MethodPotassium oxazine （350 mg·kg-1） and adenine （80 mg·kg-1） were used to construct an HUA model of mice in vivo to evaluate the mechanism related to UA reduction and the protective effect of renal function of MHD-80. Seventy male ICR mice were randomly divided into seven groups， including the normal group， model group， allopurinol group （5 mg·kg-1）， febusotan group （5 mg·kg-1）， and MHD-80 low-， medium-， and high-dose groups （3， 6， 12 mg·kg-1）， with 10 in each group. Except for the normal group， the other groups were given intragastric administration of potassium oxazine and adenine for 14 consecutive days to establish the HUA model. On the 8th to 14th day after modeling， each group was given corresponding drugs by intragastric administration， once a day. 1 h after the last administration， blood was collected from the eyeballs， and kidney and liver tissues of mice were collected. Serum levels of UA， urea nitrogen （BUN）， and creatinine （Cr） and liver activity of xanthine oxidase （XOD） were determined by enzyme colorimetry. Serum contents of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） were determined by enzyme-linked immunosorbent assay （ELISA）. Hematoxilin-eosin （HE） staining was used to observe the pathological changes in kidney tissues. The protein expression levels of ATP-binding box transporter G2 （ABCG2） and glucose-facilitating transporter 9 （GLUT9） in kidney tissues were detected by Western blot. ResultIn vivo experiment shows that compared with the normal group， the serum levels of UA， Cr， BUN， inflammatory factors TNF-α， IL-1β， and liver XOD activity in the serum of mice in the model group were significantly increased （P<0.05， P<0.01）， and the expression of GLUT9 in kidney tissues was significantly up-regulated （P<0.05）. ABCG2 protein expression was significantly down-regulated （P<0.05）， and renal injury was obvious. Compared with the model group， the levels of UA， BUN， Cr， TNF-α， IL-1β， and liver XOD activity in the serum of mice in the high-dose group of MHD-80 were decreased to different degrees （P<0.05， P<0.01）， GLUT9 protein expression was significantly down-regulated （P<0.01）， ABCG2 protein expression was significantly up-regulated （P<0.05） in the high-dose group of MHD-80， and the degree of renal injury was reduced. ConclusionMHD-80 has certain uric acid reduction， anti-inflammatory， and anti-renal injury effects， which are related to inhibiting XOD activity and regulating the expression of ABCG2 and GLUT9 uric acid transporter.

Alcohol Extract of Angelicae Pubescentis Radix Treats Gouty Arthritis Induced by MSU via NLRP3/ASC/Caspase-1 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo explore the efficacy and mechanism of the alcohol extract DH50 of Angelicae Pubescentis Radix in treating gouty arthritis induced by monosodium urate （MSU） crystals in vivo and in vitro. MethodFifty male SD rats were randomly assigned into five groups （n=10）： a normal group， a model group， a dexamethasone （DXMS， 0.07 mg·kg-1） group， and low- （DH50-D， 9 mg·kg-1） and high-dose （DH50-G， 18 mg·kg-1） DH50 groups. The rats in the normal group and model group were administrated with the same amount of pure water. On day 5， the gouty arthritis model was established by injecting MSU into the right ankle joint of rats. The toe volume and joint inflammation index were measured 4， 8， 24， and 48 h after modeling. The pathological changes of the synovial tissue were detected by hematoxylin-eosin （HE） staining. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of tumor necrosis factor-alpha （TNF-α）， interleukin （IL）-1β， and IL-6 in the synovial tissue. Western blot was employed to measure the protein levels of NOD-like receptor protein 3 （NLRP3）， cysteine-aspartic protease-1 （Caspase-1）， apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain （ASC）， IL-1β， and cyclooxygenase-2 （COX-2） in the synovial tissue. Furthermore， the cell inflammation model was established with RAW264.7 cells stimulated with MSU （75 mg·L-1）. The cell experiments were carried out with 6 groups： a normal group， a model group， a positive drug （DXMS， 100 μmol·L-1） group， and low- （DH50-D， 25 mg·L-1）， medium- （DH50-Z， 50 mg·L-1）， and high-dose （DH50-G， 100 mg·L-1） DH50 groups. Methyl thiazolyl tetrazolium （MTT） assay was employed to determine the cell viability， ELISA to determine the content of TNF-α in the supernatant of cell culture， and Western blot to determine the protein levels of NLRP3， cleaved Caspase-1， IL-1β， TNF-α， and COX-2. ResultCompared with the normal group， the rat model group showed increased toe swelling degree and joint inflammatory index （P<0.01）， serious infiltration of the synovium， elevated levels of inflammatory cytokines in the tissue homogenate （P<0.01）， and up-regulated protein levels of NLRP3， Caspase-1， ASC， IL-1β， and COX-2 （P<0.05， P<0.01）. Compared with the rat model group， low- and high-dose DH50 mitigated the toe swelling degree， decreased the joint inflammatory index， alleviated the inflammatory infiltration， lowered the levels of inflammatory cytokines in the tissue homogenate （P<0.01）， and down-regulated the expression of related proteins （P<0.05， P<0.01）. Compared with the normal group， the cell model group showed elevated level of TNF-α in the supernatant （P<0.01） and up-regulated protein levels of NLRP3， cleaved Caspase-1， IL-1β， TNF-α， and COX-2 （P<0.05）. Compared with the model group， low， medium， and high doses of DH50 lowered the level of TNF-α in the supernatant of cell culture in a dose-dependent manner and down-regulated the expression of related proteins （P<0.05， P<0.01）. ConclusionDH50 can mitigate gouty arthritis both in vitro and in vivo by inhibiting the activation of NLRP3 inflammasomes and the production of inflammatory cytokines.

Mechanism of n-butanol alcohol extract of Baitouweng Decoction in treatment of vulvovaginal candidiasis based on negative regulation of NLRP3 inflammasome via PKCδ/NLRC4/IL-1Ra axis / 中国中药杂志

This study aimed to explore the mechanism of n-butanol alcohol extract of Baitouweng Decoction(BAEB) in the treatment of vulvovaginal candidiasis(VVC) in mice based on the negative regulation of NLRP3 inflammasome via PKCδ/NLRC4/IL-1Ra axis. In the experiment, female C57BL/6 mice were divided randomly into the following six groups: a blank control group, a VVC model group, high-, medium-, and low-dose BAEB groups(80, 40, and 20 mg·kg~(-1)), and a fluconazole group(20 mg·kg~(-1)). The VVC model was induced in mice except for those in the blank control group by the estrogen dependence method. After modeling, no treatment was carried out in the blank control group. The mice in the high-, medium-, and low-dose BAEB groups were treated with BAEB at 80, 40, and 20 mg·kg~(-1), respectively, and those in the fluconazole group were treated with fluconazole at 20 mg·kg~(-1). The mice in the VVC model group received the same volume of normal saline. The general state and body weight of mice in each group were observed every day, and the morphological changes of Candida albicans in the vaginal lavage of mice were examined by Gram staining. The fungal load in the vaginal lavage of mice was detected by microdilution assay. After the mice were killed, the degree of neutrophil infiltration in the vaginal lavage was detected by Papanicolaou staining. The content of inflammatory cytokines interleukin(IL)-1β, IL-18, and lactate dehydrogenase(LDH) in the vaginal lavage was tested by enzyme-linked immunosorbent assay(ELISA), and vaginal histopathology was analyzed by hematoxylin-eosin(HE) staining. The expression and distribution of NLRP3, PKCδ, pNLRC4, and IL-1Ra in vaginal tissues were measured by immunohistochemistry(IHC), and the expression and distribution of pNLRC4 and IL-1Ra in vaginal tissues were detected by immunofluorescence(IF). The protein expression of NLRP3, PKCδ, pNLRC4, and IL-1Ra was detected by Western blot(WB), and the mRNA expression of NLRP3, PKCδ, pNLRC4, and IL-1Ra was detected by qRT-PCR. The results showed that compared with the blank control group, the VVC model group showed redness, edema, and white secretions in the vagina. Compared with the VVC model group, the BAEB groups showed improved general state of VVC mice. As revealed by Gram staining, Papanicolaou staining, microdilution assay, and HE staining, compared with the blank control group, the VVC model group showed a large number of hyphae, neutrophils infiltration, and increased fungal load in the vaginal lavage, destroyed vaginal mucosa, and infiltration of a large number of inflammatory cells. BAEB could reduce the transformation of C. albicans from yeast to hyphae. High-dose BAEB could significantly reduce neutrophil infiltration and fungal load. Low-and medium-dose BAEB could reduce the da-mage to the vaginal tissue, while high-dose BAEB could restore the damaged vaginal tissues to normal levels. ELISA results showed that the content of inflammatory cytokines IL-1β, IL-18, and LDH in the VVC model group significantly increased compared with that in the blank control group, and the content of IL-1β, IL-18 and LDH in the medium-and high-dose BAEB groups was significantly reduced compared with that in the VVC model group. WB and qRT-PCR results showed that compared with the blank control group, the VVC model group showed reduced protein and mRNA expression of PKCδ, pNLRC4, and IL-1Ra in vaginal tissues of mice and increased protein and mRNA expression of NLRP3. Compared with the VVC model group, the medium-and high-dose BAEB groups showed up-regulated protein and mRNA expression of PKCδ, pNLRC4, and IL-1Ra in vaginal tissues and inhibited protein and mRNA expression of NLRP3 in vaginal tissues. This study indicated that the therapeutic effect of BAEB on VVC mice was presumably related to the negative regulation of NLRP3 inflammasome by promoting PKCδ/NLRC4/IL-1Ra axis.

Butyl alcohol extract of Baitouweng Decoction alleviates vulvovaginal candidiasis in mice by downregulating NLRP3 inflammasome and related signal pathways / 中国中药杂志

This study aims to explore the effect of butyl alcohol extract of Baitouweng Decoction(BAEB) on vulvovaginal candidiasis(VVC) in mice and to clarify the mechanism from Toll-like receptors(TLRs)/MyD88 and Dectin-1/Syk signal pathways and NLRP3 inflammasome. To be specific, female KM mice were randomized into control group(i.g., normal saline), model group, fluco-nazole group(i.g., 20 mg·kg~(-1)), and low-dose, medium-dose, and high-dose BAEB groups(i.g., 20, 40, and 80 mg·kg~(-1), respectively). VVC was induced in mice except the control group. After the modeling, administration began and lasted 7 days. The ge-neral conditions and body weight of mice were recorded every day. On the 1 st, 3 rd, 7 th, and 14 th after vaginal infection by Candida albicans, the fungal load in the vaginal lavage fluid of the mice was measured with the plate method, and the morphology of C. albicans in vaginal lavage fluid was observed based on Gram staining. After the mice were killed, vaginal tissues were subjected to hematoxylin-eosin(HE) staining and periodic acid-Schiff(PAS) staining for vaginal histopathological analysis. The content of cytokines in vaginal lavage fluid, such as interleukin(IL)-1β, IL-18, tumor necrosis factor-α(TNF-α), IL-6, and S100 a8, was determined by enzyme-linked immunosorbent assay(ELISA), and content of reactive oxygen species(ROS) in vaginal tissues by tissue ROS detection kit. The protein expression of NLRP3, ASC, caspase-1, Dectin-1, Syk, MyD88, TLR2, TLR4, and nuclear factor-κB(NF-κB) in vaginal tissues was detected by Western blot, and the levels and distribution of NLRP3, Dectin-1, Syk, MyD88, TLR2, and TLR4 in vaginal tissues were determined with the immunohistochemical method. The results show that BAEB can improve the general conditions of VVC mice, reduce the fungal load and C. albicans hyphae in vaginal secretion, decrease ROS content in vaginal tissues and content of cytokines in vaginal lavage fluid, and down-regulate the expression of NLRP3, ASC, caspase-1, Dectin-1, Syk, MyD88, TLR2, TLR4, and NF-κB in vaginal tissues. The above results indicate that BAEB exerts therapeutic effect on VVC mice by down-regulating the key proteins in the TLRs/MyD88 and Dectin-1/Syk signal pathways and NLRP3 inflammasome.

Effect of Alcohol Extract of Spore Powder of Lasiosphaera Calvatia on Wound Healing of Full-thickness Skin Defect in Mice / 中国实验方剂学杂志

ObjectiveTo compare the effect of different solvent extracts of spore powder and fruiting body of Lasiosphaera Calvatia on fibroblasts and wound healing of full-thickness skin defect， in order to screen the optimal medication part of Lasiosphaera Calvatia. MethodThe effect of water extract and alcohol extract of spore powder and fruiting body on cell proliferation and cell migration of mouse skin fibroblasts （MSF） were examined in vitro. Cell proliferation and activity test （CCK-8） method was used for cell proliferation， scratch assay was used for cell migration， flow cytometry was conducted to explore cell cycle， enzyme-linked immunosorbent assay （ELISA） was used to determine the production of collagen Ⅰ and Ⅲ. At the same time， a full-thickness skin defect wound model was established to investigate the therapeutic effect of different solvent extracts of spore powder. Ultraviolet-visible spectrophotometry was used to measure the contents of index components in different solvent extracts. ResultThe water extract of spore powder and fruiting body had certain cytotoxicity， while the alcohol extract could promote proliferation， migration and production of collagen Ⅰ and Ⅲ of MSF， and the effect of spore powder was significantly higher than that of fruiting body. When the concentration was 10 mg·L-1， the cell proliferation rate of alcohol extract of spore powder was as high as （159.22±15.95）%， and could promote MSF from the G0/G1 phase to S phase and G2/M phase with an increased proliferation index. The alcohol extract also promoted the migration of fibroblasts， secreted collagen Ⅰ and Ⅲ. On in vivo model， the alcohol extract of spore powder significantly accelerated wound healing on mice， effectively promoted the complete epithelialization of wound tissue， and generated new collagen fiber. The results of determination showed that the contents of polyphenols and flavonoids in the alcohol extract were higher than the alcohol extract of fruiting body. ConclusionThe alcohol extract of spore powder in Lasiosphaera Calvatia has active components in the treatment of wounds with good development prospect， and the medicinal components may be polyphenols and flavonoids.

Alcohol extract of morinda officinalis improves lipid metabolism and bone metabolism in ovariectomized obese rats / 中国组织工程研究

BACKGROUND: Previous studies have showed that the alcohol extract of Morinda officinalis can effectively improve the bone quality and body mass of obese rats after ovariectomy. However, the exact mechanism is unclear. In this study, leptin and leptin receptor were used as the breakthrough point to investigate the effect of alcohol extract of Morinda officinalis on lipid metabolism and bone metabolism in ovariectomized obese rats. OBJECTIVE: To investigate the effects of alcohol extract of Morinda officinalis lipid metabolism and bone metabolism in ovariectomized obese rats. METHODS: A total of 160 SPF female Sprague-Dawley rats were randomly divided into an osteoporosis group (n=120) and a sham operation group (n=40). A postmenopausal osteoporosis model was made in the osteoporosis group by removing both ovaries. After modeling, rats in the osteoporosis group were randomly subdivided into a normal diet group, a high-fat diet group and a high-fat diet + Morinda officinalis alcohol extract group, with 40 rats in each group. The sham operation group and the normal diet group were fed with ordinary diet, while the high-fat diet group and the high-fat diet + Morinda officinalis alcohol extract group were fed with high-fat diet. The high-fat diet + Morinda officinalis alcohol extract group was gavaged with 20 g/kg Morinda officinalis alcohol extract once a day, and the remaining three groups were gavaged with 2 mL of normal saline. The study protocol was approved by the Animal Ethic Committee of Fuzhou Second Hospital of Xiamen University in September 2018 with an approval No. 20180019. RESULTS AND CONCLUSION: Compared with the sham operation group, serum leptin, leptin receptor, osteoprotegerin and high-density lipoprotein cholesterol levels were significantly lower in ovariectomized rats (P < 0.05), whereas osteocalcin, RANKL, tartrate-resistant acid phosphatase 5b, cholesterol, triacylglycerol, and low-density lipoprotein cholesterol levels were significantly higher in ovariectomized rats (P < 0.05). Compared with the high-fat diet group, serum leptin, leptin receptor, osteoprotegerin and high-density lipoprotein cholesterol levels were significantly increased in the high-fat diet + Morinda officinalis alcohol extract group (P < 0.05), whereas osteocalcin, RANKL, tartrate-resistant acid phosphatase 5b, cholesterol, triacylglycerol, and low-density lipoprotein cholesterol levels were decreased to different extents in the high-fat diet + Morinda officinalis alcohol extract group (P < 0.05). To conclusion, the alcohol extract of Morindus officinalis can up-regulate the leptin and leptin receptor expression in serum of ovariectomized obese rats, so as to improve the abnormal bone metabolism and lipid metabolism in ovariectomized obese rats.

Effect of butyl alcohol extract of Baitouweng Decoction on vaginal mucosal neutrophil chemotaxis in vulvovaginal candidiasis mice / 中国中药杂志

To investigate the effects of butyl alcohol extract of Baitouweng Decoction(BAEB) on neutrophil chemotaxis in vaginal mucosa of mice with vulvovaginal candidiasis(VVC). Seventy-two SPF female Kunming mice were randomly divided into normal control group, model group, fluconazole group, BAEB low-dose group, middle-dose group and high-dose group. Subcutaneous injection of estradiol benzoate was conducted to induce pseudo-estrus, and then 2×10~6 CFU·mL~(-1)of Candida albicans was inoculated into vaginal lumen, followed by drug treatment for 7 days. Gram staining was used to observe the morphological changes of C. albicans in vagina; vaginal fungal load was detected on agar plate. Histological changes of vaginal tissues in mice were observed by HE staining. Lactate dehydrogenase(LDH), interleukin-6(IL-6) and tumor necrosis factor(TNF-α) levels in mouse lavage fluid were detected by enzyme-linked immunosorbent assay(ELISA). Neutrophils in vaginal lavage fluid was observed and counted by using Pap smear. The levels of IL-8 and MIP-2 in vaginal mucosa were detected by ELISA. IL-8 and MIP-2 mRNA levels in vaginal mucosa of mice were detected by qRT-PCR. The results showed that as compared with the normal group, VVC model group had a large number of hyphae and a high level of fungal loadinvagina. The vaginal mucosa was completely destroyed, the number of neutrophils increased, and the protein and mRNA levels of IL-8 and MIP-2 were up-regulated. After BAEB treatment, the hyphae of the treatment group was decreased, the fungal load was decreased, the impaired mucosa showed different degrees of improvement, the inflammatory factors were decreased to varying degrees, and the protein and mRNA levels of chemokine IL-8 and MIP-2 were down-regulated. In conclusion, BAEB may be used to treat VVC by inhibiting vulvovaginal candidiasis via blocking neutrophils recruitment into vagina.

Study on material basis of Açaí cold drug based on neuro-endocrine-immune network / 中国中药杂志

To investigate the characteristics of the cold and heat properties of each resolution component of Açaí and the material basis of cooling by observing the effect of resolution components, such as Açaí oil, alcohol extract and water extract, on the neurotransmitter, endocrine hormone and immune factor level in mice with deficiency-heat and deficiency-cold syndrome. KM male mice were randomly divided into 12 groups, namely blank group, deficiency-heat model group, deficiency-heat+Açaí group, deficiency-heat+Açaí oil group, deficiency-heat+Açaí alcohol extract group, deficiency-heat+Açaí water extract group, deficiency-cold model group, deficiency-cold+Cinnamomi Cortex group, deficiency-cold+Açaí group, deficiency-cold+Açaí oil group, deficiency-cold+Açaí alcohol extract group, and deficiency-cold+Açaí water extract group. The mice in deficiency-heat group were given with thyroid tablet solution(160 mg·kg~(-1)), and the mice in deficiency-cold group were given with hydrocortisone solution(25 mg·kg~(-1)) by intragastric administration every afternoon for 14 days. The mice in each administration group received corresponding drug. The neurotransmitter, endocrine hormone and immune factor levels in the mice were measured after the experiment. The Açaí alcohol extract, consistent with the Açaí powder, showed a regulatory effect on the deficiency-heat model mice; Açaí oil and its water extract were consistent with Cinna-momi Cortex, showing a regulatory effect on the deficiency-cold model mice. In this study, on the basis of proving that Açaí was was cool in property, it also revealed that alcohol extract of Açaí was cool while oil and water extract were warm in property based on the effect of Açaí on neuro-endocrine-immune network. The results suggested that the medicine property of Açaí was the result of the comprehensive action of the resolution components with different properties, and the alcohol extract of Açaí was proved as the material basis of Açaí cold medicine by using the methods of homogeneous comparison and heterogeneous disproval.

Effect of Açaí oil, alcohol extract and water extract on temperature tendency and metabolic level of mice with deficiency-feat and deficiency-cold / 中国中药杂志

To investigate the effect of resolution components, such as Açaí oil, alcohol extract and water extract, on the temperature tendency animal behavior and intrinsic biochemical indexes, such ascyclic nucleotides and metabolic level, in mice with deficiency-heat and deficiency-cold syndrome, in order to study the characteristics of the cold and heat properties of each resolution component of Açaí and the material basis of cooling. KM mice were randomly divided into 12 groups, namely blank group, deficiency-heat model group, deficiency-heat+Açaí group, deficiency-heat+Açaí oil group, deficiency-heat+Açaí alcohol extract group, deficiency-heat+Açaí water extract group, deficiency-cold model group, deficiency-cold+Cinnamomi Cortex group, deficiency-cold+Açaí group, deficiency-cold+Açaí oil group, deficiency-cold+Açaí alcohol extract group, deficiency-cold+Açaí water extract group. The mice in deficiency-heat group were given thyroid tablet solution(160 mg·kg~(-1)), the mice in deficiency-cold group were given hydrocortisone solution(25 mg·kg~(-1)) through gastric perfusion every afternoon for 14 days, and each administration group was given the corresponding drug. The temperature tendency, cyclic nucleotides and metabolic level of animals were measured after the experiment. The Açaí alcohol extract was consistent with the Açaí powder, with a regulatory effect on the deficiency-heat model mice; Açaí oil and its water extract were consistent with Cinnamomi Cortex, with a regulatory effect on the deficiency-cold model mice. In this study, based on the parable theory of traditional Chinese medicine's properties and tastes, property of alcohol extract of Açaí was cool, while the property of oil and water extract were warm, the alcohol extract of Açaí was the material basis of Açaí cold medicine by the methods of homogeneous comparison and heterogeneous disproval.

Effect of Chaenomelis Fructus Alcohol Extract on Arthritis Inflammation and Apoptosis of Synovial Cells in Rheumatoid Arthritis Model / 中国实验方剂学杂志

Objective:To study the protective effect and mechanism of Chaenomelis Fructus alcohol extract (CFE) on the synovium of rheumatoid arthritis (RA). Method:Sixty male SD rats were randomly divided into normal group and model group. RA model was made by injection of complete Freund's adjuvant， and then was randomly divided into model group， CFE low， medium and high dose group and Tripterygium glycoside group according to the inflammatory score. The CFE groups (0.15，0.30，0.60 g·kg-1·d-1) had intragastric administration once a day for 30 d after the model establishment. The blank control group and the model group were given the same volume saline water by gavage. After all the drugs were given， the blood， joint tissues and synovium tissue of rats were collected. The levels of interleukin-1β (IL-1β)， interleukin-6 (IL-6)， interleukin-10 (IL-10) and tumor necrosis factor (TNF)-α in serum were detected by enzyme linked immunosorbent assay (ELISA)， the pathological changes of synovium were observed by hematoxylin eosin (HE) staining， and the expressions of B-cell lymphoma-2 (Bcl-2)，Bcl-2 associated X protein (Bax) and death factor (Fas) in joints were detected by Western blot. Result:Compared with normal group， the swelling degree and inflammation index of rats' feet in model group increased significantly， the levels of inflammatory factors IL-1β， IL-6， TNF-α in serum increased (P<0.01)， anti-inflammatory factor IL-10 decreased (P<0.01)， the protein expression of Bax， Fas and Bcl-2 increased， and the statistical results of Bcl-2 showed significant difference (P<0.05). Compared with model group， the swelling degree and inflammatory index of the plantar of RA rats were improved in the middle and high dose groups of CFE (P<0.01)， the pathological changes such as synovial tissue hyperplasia and inflammatory cell infiltration were reduced in each dose group， and the levels of IL-1β， IL-6 and TNF-α in serum were reduced (P<0.05)， anti inflammatory factor IL-10 increased (P<0.05)， the disorder of inflammatory cytokine in the model was corrected， Bax， Fas expression increased， Bcl-2 protein expression decreased (P<0.01). Conclusion:CFE can reduce the degree of inflammation in RA joint and has obvious anti RA effects， which may be related to the apoptosis of synoviocytes induced by CFE.

Quality Evaluation of Curcumae Radix Introduced in Zhongshan by Gray Relational Model / 中国实验方剂学杂志

Objective:To establish the gray relational analysis for quality evaluation of the samples of Curcumae Radix introduced in Zhongshan. Method:With volatile oil and curcumin as Q-markers,and alcohol extract,germacrone,germacr-1(10)-ene-5,8-dione and curcumin as comprehensive evaluation index, the contents of the four main components in 72 samples of Curcumae Radix of 3 different varieties introduced in Zhongshan from 3 different regions were determined. The grey relational method was used to build the gray correlation evaluation model for Curcumae Radix introduced in Zhongshan. Result:The relative correlation degree (γi) of 72 samples was between 0.262 and 0.697,in which γi was above 0.450 for 10 samples,and below 0.300 for 37 samples,indicating great differences in the quality of Curcumae Radix after introduction. The γi was 0.697 and 0.525 respectively for No.MY-W-4 and No.MY-W-1 from Curcumae Radix in Mayu with the best quality. The average values of γi for the samples of 3 different varieties from 3 different regions were between 0.281 and 0.420,and Mayu samples had the maximum average value,indicating that Mayu samples had the highest overall quality of,and could be introduced as excellent resources. Conclusion:The evaluation method combined with GRA method and multi-index quantification was simple,objective and comprehensive, and could be used to evaluate the quality of Curcumae Radix introduced in Zhongshan,so as to provide references for screening high-quality provenance.

Isolation and Identification of Chemical Constituents from Cinnamomi Ramulus / 中国实验方剂学杂志

Objective:The research group found in the early stage that the 75%alcohol extract of the Cinnamomi Ramulus had a significant physiological activity in inhibiting necroptosis by screening out the self-built sample library of 100 kinds of traditional Chinese medicines in Jiangxi. To identify the active components and find the target compounds,the 75%alcohol extracts of Cinnamomi Ramulus were isolated and studied systemically in chemistry. Method:The 20 kg dry Cinnamomi Ramulus was crushed into coarse powder,and extracted with 75%alcohol for four times, one time every 7 d. Then total extracts were obtained after solvent was recycled under decompression. The extract was separated by D101 macroporous resin column chromatography and eluted by water,30%ethanol,50%ethanol,70%ethanol,90%ethanol,so as to get the corresponding fraction finally. The compounds in the 30%ethanol and 50%ethanol fraction were isolated and purified by chromatography on silica gel,Sephadex LH-20 column and high pressure preparative chromatography,and their structures were determined according to physicochemical properties and spectral analysis. Result and Conclusion:Thirteen compounds were isolated and identified as (+)-syringaresinol (1),(+)-lyoniresinol (2),spicatolignan B (3),(-)-secoisolariciresinol (4),ovafolinin B (5),protocatechualdehyde (6),protocatechuic acid (7),syringaldehyde (8),vanillic acid (9),ethyl protocatechuate (10),syringic acid (11),ethyl gallate (12),2-(3',4'-dihydroxyphenyl)-1,3-pepper ring-5-aldehyde (13). Compounds 1-5,10-13 were isolated from this plant for the first time.

Phenolic Components from Roots of Psidium guajava / 中国实验方剂学杂志

Objective:The different parts of Psidium guajava(Myrrhinaceae) have different bioactivities. There are intensive studies for chemical constituents of its leaves and fruits at present. However,there are a few studies for the roots. Therefore, we systematically investigated the chemical constituents of phenolic acids in roots of P.guajava. Method:The chemical constituents were isolated and purified by various column chromatography methods and semi-preparative HPLC. The isolated compounds were identified by physicochemical properties and spectral analysis. Result:Sixteen phenolic components were isolated from the ethanol extract and identified as 3,3',4'-tri-O-methylellagic acid(1),3,3',4-tri-O-methylellagic acid-4'-O-β-D-glucopyranoside(2),3-O-methylellagic acid-4'-O-α-L-rhamnopyranoside(3),3'-O-methyl-3,4-O,O-metheneellagic acid-4'-O-β-D-glucopyranoside(4),gallic acid(5),methyl gallate(6),ethyl gallate(7), 3,4,5-trimethoxypheny-1-β-D-glucopyranoside(8),3,5-dimethoxy-4-hydroxy-benzoic acid-7-O-β-D-glucoside(9),1-hydroxy-3,4,5-trimethoxyphenyl-1-O-[6'-O-(4″-carboxy-1″,3″,5″-trihydroxy)phenyl]-β-D-glucopyranoside(10),vanillic acid(11),protocatechuic acid(12),secoisolariciresinol 9-O-β-D-glucopyranoside(13),phloretin 4'-O-β-D-glucopyranoside(14),cinchonain Ib(15) and epicatechin(16). Conclusion:Compounds 3,4,6,8-10,13-15 were isolated from this plant for the first time.

Mechanism of butyl alcohol extract of Baitouweng Decoction (BAEB) on Candida albicans biofilms based on pH signal pathway / 中国中药杂志

This study aimed to investigate the effect of butyl alcohol extract of Baitouweng Decoction( BAEB) on Candida albicans biofilms based on pH signal pathway. The morphology of biofilms of the pH mutants was observed by scanning electron microscope. The biofilm thickness of the pH mutants was measured by CLSM. The biofilm activity of the pH mutants was analyzed by microplate reader.The biofilm damage of the pH mutants was detected by flow cytometry. The expression of pH mutant biofilm-related genes was detected by qRT-PCR. The results showed that the deletion of PHR1 gene resulted in the defect of biofilm,but there were more substrates for PHR1 complementation. BAEB had no significant effect on the two strains. RIM101 gene deletion or complementation did not cause significant structural damage,but after BAEB treatment,the biofilms of both strains were significantly inhibited. For the biofilm thickness,PHR1 deletion or complementation caused the thickness to decrease,after BAEB treatment,the thickness of the two strains did not change significantly. However,RIM101 gene deletion or complementation had little effect on the thickness,and the thickness of the two strains became thinner after adding BAEB. For biofilm activity,PHR1 deletion or complementation and RIM101 deletion resulted in decreased activity,RIM101 complementation did not change significantly; BAEB significantly inhibited biofilm activity of PHR1 deletion,PHR1 complemetation,RIM101 deletion and RIM101 complemetation strains. For the biofilm damage,PHR1 gene deletion or complementation,RIM101 gene deletion or complementation all showed different degrees of damage; after adding BAEB,the damage rate of PHR1 deletion or complementation was not significantly different,but the damage rate of RIM101 deletion or complementation was significantly increased. Except to the up-regulation of HSP90 gene expression,ALS3,SUN41,HWP1,UME6 and PGA10 genes of PHR1 deletion,PHR1 complementation,RIM101 deletion,and RIM101 complementation strains showed a downward expression trend. In a word,this study showed that mutations in PHR1 and RIM101 genes in the pH signaling pathway could enhance the sensitivity of the strains to the antifungal drug BAEB,thus inhibiting the biofilm formation and related genes expression in C. albicans.

Effects of different drying methods on appearance and internal components of Notoginseng Radix et Rhizoma / 中草药

Objective: The effects of different drying methods (drying-in-the-shade, sun-drying, freeze-drying and hot air drying) on the appearance traits, internal structure and quality of the main roots of Notoginseng Radix et Rhizoma (NRR) were revealed, which provided a theoretical basis for screening the drying methods suitable for the primary processing of NRR. Methods: In this study, the effect of four different drying methods on drying rate, rehydration rate, appearance traits, alcohol extracts and internal components (dencichine, saponin component notoginsenoside R1 and ginsenoside Rg1, Rb1, Rd, Re, and reducing sugar, total sugar) of NRR were compared. Results: Fresh NRR was dried by drying-in-the-shade method, and the drying rate was slow, which took about 473 h. However, the appearance quality of the medicinal materials was excellent with firm texture, slow rehydration rate, and high content of total saponins and dencichine. The drying rate of materials under sun drying method was also slow, and due to the long drying cycle, the starch and sugar of the medicinal materials were more decomposed, resulting in the whitening part of the medicinal materials of NRR. When fresh NRR was dried by hot air, the drying rate was faster and the time was shortened. When the temperature was 40 ℃, the appearance of the medicine was not much different from that of the drying-in-the-shade method. After drying, the material was firmer, except for the content of dencichine, the content of saponin had no difference between that of drying-in-the-shade; Due to the high drying temperature at 50 ℃ and 60 ℃, the excessive dehydration rate led to the hollowness of the NRR, and medicinal material was not solid; The rehydration rate was fast, and the content of NRR spilled out, resulting in sugar coking, and the color of the cross section of the medicinal material changed into deeper with the increase of the drying temperature, which resulted in the decomposition of starch, and the significant increase of total sugar and reducing sugar content. The lyophilized medicinal material had a very fast rehydration rate, and the internal texture was loose and porous, and the texture became light, but the saponin component and dencichine were the highest active ingredients. Conclusion: Considering the aspects of appearance, medicinal ingredients and cost, fresh NRR dried by drying-in-the-shade method obtained solid material, compacted internal structure, good appearance and high content of medicinal ingredients. The primary processing method of NRR should be drying-in-the-shade. If the processing volume of the medicinal material is large, it needs to shorten the drying time, and the primary processing method of NRR should be hot air drying process at about 40 ℃. If the high-content medicinal ingredients are the purpose, it is recommended to use freeze-drying.

Effects of butyl alcohol extract of Baitouweng Decoction on Candida albicans adhesion based on pH-mutant strains under acidic conditions / 中草药

Objective: To investigate the effect and mechanism of butyl alcohol extract of Baitouweng Decoction (BAEB) on adhesion of Candida albicans based on pH signaling pathway. Methods: Spot assay method was used to detect the sensitivity of pH mutants to BAEB under acidic conditions. XTT assay was used to detect the effect of BAEB on metabolic activity of pH mutants. The effect of BAEB on the adhesion activity of pH mutants was observed by fluorescence microscopy. The effect of BAEB on hydrophobicity of pH mutant was determined by n-octane inclusion method. The effect of BAEB on the expression of adhesion genes related to pH mutants was detected by qRT-PCR. Results: Under acidic conditions, spot assay observation showed that pH mutants were less sensitive to BAEB, 512 μg/mL BAEB interfered with pH mutants for 24 h and 48 h, there was no significantly decrease in bacterial colony. XTT assay showed that the metabolic activity of WT, PHR2 complementation, rim101/rim101 and RIM101 complementation was significantly inhibited in 512 μg/mL BAEB, and there was no significantly difference in the inhibition of phr2/phr2 metabolic activity. Fluorescence microscopy showed that the cell adhesion activity of WT, PHR2 complementation, rim101/rim101, RIM101 complementation was significantly inhibited in 512 μg/mL BAEB, the cell adhesion activity of phr2/phr2 had no obvious effect in 512 μg/mL BAEB. The n-octane inclusion method showed that the effect of 512 μg/mL BAEB on the cell surface hydrophobicity of WT, phr2/phr2, PHR2 complementation, rim101/rim101, RIM101 complementation was not significant. The qRT-PCR assay showed that the adhesion genes of pH mutants was inhibited in 1024 μg/mL BAEB. Conclusion: Under acidic conditions, the Candida albicans pH mutants was inhibited by BAEB to a certain extent.

Effect of Kadsura coccinea Alcohol Extract on Immunologic Hepatic Fibrosis In Rats / 中国药学杂志

OBJECTIVE: To study the effects of Kadsura coccinea alcohol extract(KCAE) on rats with immunologic hepatic fibrosis and research the possible mechanisms in it. METHODS: Totally 60 SD male rats were randomly divided into 6 groups: a normal control group,a model group, a compound Biejia-ruangan tablets group(0.7 g•kg-1), KCAE high, middle and low dose groups(1.68, 0.84, 0.42 g•kg-1) at ten in each groups. Except for the normal control group,other groups were duplicated intraperitoneal injection of porcine serum twice a week at dose of 0.5 mL•time-1. The rats in treatment groups were intragastric administration respectively, meanwhile, the rats in normal control and model groups were treated with the same volume of distilled water, once a day for 15 weeks. The liver was weighed to calculate the liver index. Alanine aminotransferase(ALT), aspartate aminotransferase(AST), total protein(TP), albumin(ALB) and total bilirubin(TB) were evaluated by the Mind-Ray automatic biochaemical analyzer. The expression level of procollagen III(PCIII), collagen type (-C), laminin(LN), hyaluronic acid(HA), transforming growth factor-β1(TGF-β1), interkeukin-10(IL-10), interferon-γ(IFN-γ) and tumor necrosis factor-α(TNF-α) in serum were detected by ELISA. The degrees of fibrosis were evaluated by HE and Masson straining, and the expression levels of TGF-β1 in liver tissue were assessed by Western blot. RESULTS: Compared with model group, the liver index of KCAE high-dose group was decreased significantly(P0.05). KCAE could decrease the level of PCIII, IV-C, LN, HA, TGF-β1, TNF-α and increase the level of IFN-γ in serum. KCAE could alleviate the hepatic fibrosis in rats(P<0.01) and inhibit the expression of TGF-β1 in the liver tissues significantly(P<0.01). CONCLUSION: KCAE has an anti-immunologic hepatic fibrosis effect in rats and the mechanisms possibly involve effectively regulating inflammatory cytokines, reducing extracellular matrix expression and inhibiting the expression of TGF-β1.

Butyl alcohol extract of Baitouweng decoction inhibits Candida albicans cell membrane / 中国中药杂志

To study the inhibitory effect of butyl alcohol extract of Baitouweng decoction(BAEB) on Candida albicans cell membrane. The effects of BAEB on the activity of C. albicans were observed by Spot assay. The changes of intracellular osmotic pressure of C. albicans after BAEB intervention were detected by microtiter plate reader. The effect of BAEB on cell membrane permeability of C. albicans were observed by fluorescence microscopy. The content of ergosterol in C. albicans cell membrane was detected by high performance liquid chromatography, and the expression of ergosterol biosynthesis related genes in cell membrane was detected by qRT-PCR. The results showed that the activity of C. albicans was significantly decreased in 256, 512 and 1 024 mg•L⁻¹ BAEB group. The intracellular glycerol content of C. albicans was significantly increased in 512 and 1 024 mg•L⁻¹ BAEB group(P<0.05). The gene HOG1 associated with intracellular osmotic pressure of C. albicans was down-regulated by 9.1, 9.3 and 5.5 times, respectively. C. albicans with red fluorescent were increased significantly in 512 and 1 024 mg•L⁻¹ BAEB group. The peak area of ergosterol in the 1 024 mg•L⁻¹ BAEB group was 35.884 95, with a significant difference(P<0.05); ERG1, ERG2, ERG3, ERG4, ERG5, ERG6, ERG10, ERG11, ERG13, ERG24, ERG25, ERG251, ERG26 and UPC2 were down-regulated by 6.58, 4.89, 4.15, 9.24,3.41, 9.84, 3.08, 7.50, 5.53, 5.90, 2.45, 3.25,1.98 and 10.07 times respectively in 1 024 mg•L⁻¹ BAEB group. The study indicated that BAEB could inhibit ergosterol and its biosynthesis related genes expression in the cell membrane and inhibit the activity of C. albicans.

Antitumor Activity of Alcohol Extract from Murraya exotica L / 中国药学杂志

OBJECTIVE: To investigate the cytotoxic activities of chemical constituents in alcohol extract of the stem bark of Murraya exotica L. METHODS: The cytotoxicity against five cancer cell lines, U937, HL-60, K562, Bel7402 and Hela, were assayed by MTT and SRB METHODS. The constituents were isolated from Murraya exotica L. by routine chromatographic METHODS and the structures of the isolates were elucidated by NMR techniques. The antitumor effect was evaluated on cancer cells in vitro. RESULTS: When the cancer cells were exposed to the extract for more than 48 h, the survival rate decreased with the increase of the drug concentration. Four compounds were isolated and identified as isolariciresinol (I), dimethoxy isolariciresinol (II), 3-methoxyisolariciresinol (III), and 5'-methoxyisolariciresinol (IV). CONCLUSION: The active ingredients in Murraya exotica L. have antitumor activities, which may be compounds I and II.