In order to improve reproduvtive efficieny and understand reproduvtive defense mechanism, the oviduct, uterine horn and uterine body of bovine were used to detect the changes of inflammatory factors and the relationsh...In order to improve reproduvtive efficieny and understand reproduvtive defense mechanism, the oviduct, uterine horn and uterine body of bovine were used to detect the changes of inflammatory factors and the relationship between estrogen and progesterone receptor protein during estrous cycle by real-time PCR and Elisa method. The results showed that interleukin-4（IL-4）, interleukin-6（IL-6）, interleukin-10（IL-10）, interleukin-1α（IL-1α） and interleukin-1β（IL-1β） were expressed in cow oviduct, uterine horn and uterine body. In the follicular phase and the luteal phase, m RNA expression of five inflammatory factors in the uterine horn and uterine body was higher than that in the oviduct. In the follicular phase, IL-10 was highly expressed in the uterine horn and uterine body, IL-4 was highly expressed in the uterine horn, uterine body and oviduct. Additionally, in the luteal phase, IL-6 and IL-1β were highly expressed in the uterine horn, uterine body and oviduct, and the highest expression of IL-1β was observed in the uterine horn. The levels of Estrogen Receptor（ERα） protein in the oviduct, uterine horn and uterine body significantly increased in the follicular phase. The levels of Progesterone Receptor（PR） protein in the same portions of the reproductive tract in the luteal phase were significantly higher than those in the follicular phase. IL-4 and IL-10 in the cow reproductive tract might play a major role in the follicular phase, while IL-6 and IL-1β might play a major role in the luteal phase. The expression of five inflammatory factors was not directly regulated by ERα and PR.展开更多
OBJECTIVE Chronic stress is one of the important factors in the development of many mental and neurological diseases,and cause damage to the central nervous system,affect animal emotions and damage the immune function...OBJECTIVE Chronic stress is one of the important factors in the development of many mental and neurological diseases,and cause damage to the central nervous system,affect animal emotions and damage the immune function of the body.The purpose of this study was to investigate the effects of LW-AFC which extracting from traditional Chinese medicine prescription Liuwei Dihuang decoctionon the anxiety-like behaviorand immune dysfunction abnormalities caused by chronic stress,and whether immune intervention affect the action of LW-AFC.METHODS Male BALB/c mice were subcutaneously injected with corticosterone(25 mg·kg^-1)for 28 d to establish a chronic stress model.Cyclophos⁃phamide(Cy,80 mg·kg^-1)was injected continuously for the initial three days,followed by once a week,LW-AFC(1.6 g·kg^-1)was given continuously for 28 d.Then investigate the emotion changes by open field and elevated plus maze tests,and detected the lymphocyte proliferation,lymphocyte subsets in peripheral blood,microglia and astrocyte expression,and inflammatory cytokines in peripheral blood and brain tissue.RESULTS The mice showed obvious depressive-like behaviorafter 28 d of continuous corticosterone injection.LW-AFC could significantly improve the anxietybehavior induced by corticosterone injection,but LW-AFC could not improve the anxietybehavior of mice by Cy intervention.The expression of glial cells in hippocampus of corticosterone-induced mice showed an upward trend,and the activation of microglia and astrocytes have significantly increase in corticosterone and Cy injected mice.LW-AFC significantly decreased the activation of microglia and astrocytes in corticosterone-induced mice with Cy intervention.This suggested that LW-AFC can reduce the damage of stress on the immune function of central nervous system under immunosuppres⁃sive state.Furthermore,LW-AFC could significantly up-regulate the proliferation of splenic lymphocyte stimulated by LPS and ConA,up-regulate the proportion of CD3+CD8+cells,reduce the proportion of CD4+/CD8+cells,decrease the secretion of inflammatory factors IL-6 and MCP-1 in plasma,and increase the level of anti-inflammatory factor IL-10 in plasma of mice induced by chronic corticosteroneinjection.While LW-AFC could promote the inflammatory factors TNF-α and IL-6in plasma,inhibit the secretion of anti-inflammatory factor IL-10 and inflammatory cytokine MCP-1 in hippocampus of corticosterone-induced mice with Cy intervention.CONCLUSION LW-AFC can improve anxiety-likebehavior induced by chronic stress,the Cy intervention affects the alleviation of anxiety-like behavior by LW-AFC as well as the regulation of immune function.The regulation of immune function might be the main way for LW-AFC to improve the function of central nervous system.展开更多
OBJECTIVE Microglial activation contributes to neuroinflammation and neuronal damages in neurodegenerative disorders including Alzheimer and Parkinson diseases.It has been suggested that neurodegenerative disorders ma...OBJECTIVE Microglial activation contributes to neuroinflammation and neuronal damages in neurodegenerative disorders including Alzheimer and Parkinson diseases.It has been suggested that neurodegenerative disorders may be improved if neuroinflammation can be controlled.Isoliquiritigenin(ISL)isolated from Glycyrrhiza glabra possess potent anti-inflammatory capability,we thus investigate the inhibitory effects of ISL on LPS-induced microglial activation and neuroinflammation and the roles of neuroprotection on neurodegenerative disorders.Moveover,we would explore the mechanism of itsassociated inflammatory signaling passway.METHODS By cultivating,isolating and purifying,we got the primary microglia of the rat.The form of those cells was observed under an optical microscope,the purity was identified by the CD11b immunefluorescence staining.Firstly,the effects of ISL on microglial viability were explored by the MTT assay.The microglial cells were pretreated by ISL(5μmol·L-1)and then stimulated by LPS(100 ng·mL-1),the production of NO was detected by Griess reagent,the change of microglial morphology was observed by immunefluorescence staining.The production of IL-1βand TNF-αin culture medium were tested by ELISA,and the m RNA expression of pro-inflammatory factors,including IL-1β,TNF-α,i NOS and COX-2 was detected by real time PCR.The protein expression of i NOS and COX-2was detected by Western blotting.RESULTS(1)After purifying,microglial showed the typical morphological characteristics.The cell body was small,protruding outstretched in the branch rod,spindle.After LPS stimulation,the shape was changed to amoeba-like,and the protrusions retracted,cell rounding.In the CD11b immunefluorescence staining,the purity of the microglial was over 98%,and showed powerful phagocytic activity.(2)The results show that ISL(0.16-20μmol·L-1)concentration has no effect on primary microglia′s vitality.And at the concentration of 5μmol·L-1ISL has the significantly inhibitory effect on NO production induced by LPS.(3)The results shows that ISL pretreatment can significantly inhibit LPS-induced microglial activation,and can significantly reduce the production of NO in a dose-dependent manner(P<0.05).When stimulated by LPS for12 h,the expression of TNF-αand IL-1βm RNA in the culture medium are reduced by ISL pre-treatment(P<0.05).After LPS stimulation for 24 h,the expression of COX2 and i NOS m RNA and protein can be reduced(P<0.05).After stimulated by LPS from 0.5 to 2 h,p-ERK expression has be decreased.CONCLUSION We successfully obtain primary microglia with high-purity by culturing in vitro.We find that ISL can significantly reduce the levels of proinflammatory facters,which indicate ISL can inhibit microglia activation and neuroinflammation by blocking ERK1/2 signal pathway.展开更多
Schisandra Chinensis Fructus(SCF)is the fruit of Schisandra chinensis(Turcz.)Baill.,a perennial vine.It was first recorded in Shen Nong′s herbal classic and has a long application history.Studies have shown that SCF ...Schisandra Chinensis Fructus(SCF)is the fruit of Schisandra chinensis(Turcz.)Baill.,a perennial vine.It was first recorded in Shen Nong′s herbal classic and has a long application history.Studies have shown that SCF has anti-inflammatory,protective liver,antioxidant,antibacterial and other pharmacological effects.Ancient prescriptions are commonly used in the treatment of chronic diarrhea and other intestinal diseases and diabetes.Modern clinical pharmacology features of SCF polysaccharide(SCFP)in diabetes,liver diseases,enteritis and other aspects have achieved excellent results.Gut is an important digestive organ of human body,but intestinal diseases are varied,including Crohn′s disease,ulcerative colitis,intestinal flora imbalance,etc..It is a chronic and non-specific inflammatory disease.The disease is persisted for a long time and the incidence rate is expected to rise.Most of the symptoms are recurrent diarrhea,bloody stool and abdominal pain.It is considered by the World Health Organization as a refractory disease.At present,there is little possibility of complete cure,which is closely related to complex environmental factors,eating habits and heredity.In recent years,clinical studies have found that SCFP has a variety of pharmacological effects on intestinal protection.①Reduce inflammatory factors:intestinal mucositis is a common adverse reaction in patients with chemotherapy.The development of mucositis is related to pro-inflammatory factors such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),IL^(-1)β,Interferon-γ(IFN-γ).SCFP can significantly reduce IL-6 TNF-α,IL^(-1)β,and IL-8,as well as the accumulation of T cells in the process of resisting apoptosis,reduce the inflammatory reaction and protect the damage to villi and crypts,improve the symptoms of small intestinal mucositis caused by weight loss and diarrhea.②Promote immunoglobulin A secretion:intestinal mucosal immunity is the first line of defense of the body′s immune system.Its main antibody is secretory immunoglobulin A,which can destroy and phagocytize microorganisms,bacteria and viruses.SCFP can improve intestinal immunity by increasing the number and activity of T lymphocytes,promoting the secretion of secretory immunoglobulin A,and affecting the activity of a variety of cytokines.③Regulation of intestinal flora:the flora in the intestine has the functions of auxiliary nutrient absorption,biological antagonism and immune regulation,and can form a natural barrier for the host's intestine.When the human intestinal flora is disordered,probiotics will be greatly reduced,harmful bacteria will proliferate and destroy the intestinal environment.Under these conditions,the intake of SCFP significantly increased the number of beneficial bacteria such as bifidobacteria and lactobacillus,and significantly decreased the number of conditional pathogens such as enterococcus and escherichia coli,indicating that SCFP can indeed regulate the intestinal disorder caused by lincomycin hydrochloride to a certain extent. This may be because beneficial bacteria in the intestine metabolize polysaccharides produce short chain fatty acids such as lactic acid and acetic acid, which reduces the pH value in the intestine and inhibits the growth of enterococcus and Escherichia coli. In conclusion, SCFP can treat and protect intestinal diseases to a certain extent, which provides a favorable basis for the treatment of intestinal diseases.展开更多
The pathogenesis of animal osteoarthritis(OA)is not clear.The aim of this study was to investigate the dynamic changes of cartilage lesions and serum IL-1β,TNF-α,IL-6 and IL-17 inflammatory factors in a post-traumat...The pathogenesis of animal osteoarthritis(OA)is not clear.The aim of this study was to investigate the dynamic changes of cartilage lesions and serum IL-1β,TNF-α,IL-6 and IL-17 inflammatory factors in a post-traumatic osteoarthritis model in rats,and to explore their values in monitoring the progression of osteoarthritis.Forty male SD rats were randomly divided into the control group(n=20)and the model group(n=20).The OA model was established by anterior cruciate ligament transection combined with partial medial meniscus resection(ACLT+PMMx)in the model group,and sham operation was performed in the control group.On the 0th,10th,20th and 30th days after modeling,the degree of joint swelling and the number of white blood cells in the peripheral blood were evaluated,and tibia samples were taken for macroscopic observation score and pathological observation.The serum levels ofIL-1β,TNF-α,IL-6 and IL-17 concentration were detected by enzyme-linked immunosorbent assay(ELISA).The results showed that the cartilage damage in the tibial surface and pathological observations of rats in the model group gradually increased with time,the eye scores increased and increased significantly on the 20th and 30th days(P<0.01),and the joints of the rats were swollen.And the swelling was extremely significant on the 30th day(P<0.01).Compared with the control group,the number of white blood cells in the peripheral blood of the model group gradually increased,and the increase was extremely significant on the 20th and 30th days(P<0.01).The serum IL-17 concentration in the model group tended to increase,and the increase was significant on the 20th day(P<0.05)and extremely significant on the 30th day(P<0.01);the serum TNF-αand IL-1βconcentrations in the model group showed an increasing trend,and the increase was extremely significant on the 10th,20th and 30th days(P<0.01);IL-6 concentration in the serum of the model group showed an increasing trend,and it increased significantly on the 30th day(P<0.05).The results showed that in the rat model of traumatic OA,the degree of joint damage gradually increased,and the expression of blood inflammatory indexes IL-1β,TNF-α,IL-17 and IL-6 increased.The research provided a basis for prompting and monitoring the timing of OA treatment and the development of the disease in animal clinical.展开更多
基金Supported by the National International Scientific and Technological Cooperation Project(2011DFA30760-2-1)
文摘In order to improve reproduvtive efficieny and understand reproduvtive defense mechanism, the oviduct, uterine horn and uterine body of bovine were used to detect the changes of inflammatory factors and the relationship between estrogen and progesterone receptor protein during estrous cycle by real-time PCR and Elisa method. The results showed that interleukin-4（IL-4）, interleukin-6（IL-6）, interleukin-10（IL-10）, interleukin-1α（IL-1α） and interleukin-1β（IL-1β） were expressed in cow oviduct, uterine horn and uterine body. In the follicular phase and the luteal phase, m RNA expression of five inflammatory factors in the uterine horn and uterine body was higher than that in the oviduct. In the follicular phase, IL-10 was highly expressed in the uterine horn and uterine body, IL-4 was highly expressed in the uterine horn, uterine body and oviduct. Additionally, in the luteal phase, IL-6 and IL-1β were highly expressed in the uterine horn, uterine body and oviduct, and the highest expression of IL-1β was observed in the uterine horn. The levels of Estrogen Receptor（ERα） protein in the oviduct, uterine horn and uterine body significantly increased in the follicular phase. The levels of Progesterone Receptor（PR） protein in the same portions of the reproductive tract in the luteal phase were significantly higher than those in the follicular phase. IL-4 and IL-10 in the cow reproductive tract might play a major role in the follicular phase, while IL-6 and IL-1β might play a major role in the luteal phase. The expression of five inflammatory factors was not directly regulated by ERα and PR.
基金National Natural Science Foundation of China(81473191)National Key Research and Development Program(2016YFC1306300)
文摘OBJECTIVE Chronic stress is one of the important factors in the development of many mental and neurological diseases,and cause damage to the central nervous system,affect animal emotions and damage the immune function of the body.The purpose of this study was to investigate the effects of LW-AFC which extracting from traditional Chinese medicine prescription Liuwei Dihuang decoctionon the anxiety-like behaviorand immune dysfunction abnormalities caused by chronic stress,and whether immune intervention affect the action of LW-AFC.METHODS Male BALB/c mice were subcutaneously injected with corticosterone(25 mg·kg^-1)for 28 d to establish a chronic stress model.Cyclophos⁃phamide(Cy,80 mg·kg^-1)was injected continuously for the initial three days,followed by once a week,LW-AFC(1.6 g·kg^-1)was given continuously for 28 d.Then investigate the emotion changes by open field and elevated plus maze tests,and detected the lymphocyte proliferation,lymphocyte subsets in peripheral blood,microglia and astrocyte expression,and inflammatory cytokines in peripheral blood and brain tissue.RESULTS The mice showed obvious depressive-like behaviorafter 28 d of continuous corticosterone injection.LW-AFC could significantly improve the anxietybehavior induced by corticosterone injection,but LW-AFC could not improve the anxietybehavior of mice by Cy intervention.The expression of glial cells in hippocampus of corticosterone-induced mice showed an upward trend,and the activation of microglia and astrocytes have significantly increase in corticosterone and Cy injected mice.LW-AFC significantly decreased the activation of microglia and astrocytes in corticosterone-induced mice with Cy intervention.This suggested that LW-AFC can reduce the damage of stress on the immune function of central nervous system under immunosuppres⁃sive state.Furthermore,LW-AFC could significantly up-regulate the proliferation of splenic lymphocyte stimulated by LPS and ConA,up-regulate the proportion of CD3+CD8+cells,reduce the proportion of CD4+/CD8+cells,decrease the secretion of inflammatory factors IL-6 and MCP-1 in plasma,and increase the level of anti-inflammatory factor IL-10 in plasma of mice induced by chronic corticosteroneinjection.While LW-AFC could promote the inflammatory factors TNF-α and IL-6in plasma,inhibit the secretion of anti-inflammatory factor IL-10 and inflammatory cytokine MCP-1 in hippocampus of corticosterone-induced mice with Cy intervention.CONCLUSION LW-AFC can improve anxiety-likebehavior induced by chronic stress,the Cy intervention affects the alleviation of anxiety-like behavior by LW-AFC as well as the regulation of immune function.The regulation of immune function might be the main way for LW-AFC to improve the function of central nervous system.
基金The project supported by National Natural Science Foundation of China(81274119)
文摘OBJECTIVE Microglial activation contributes to neuroinflammation and neuronal damages in neurodegenerative disorders including Alzheimer and Parkinson diseases.It has been suggested that neurodegenerative disorders may be improved if neuroinflammation can be controlled.Isoliquiritigenin(ISL)isolated from Glycyrrhiza glabra possess potent anti-inflammatory capability,we thus investigate the inhibitory effects of ISL on LPS-induced microglial activation and neuroinflammation and the roles of neuroprotection on neurodegenerative disorders.Moveover,we would explore the mechanism of itsassociated inflammatory signaling passway.METHODS By cultivating,isolating and purifying,we got the primary microglia of the rat.The form of those cells was observed under an optical microscope,the purity was identified by the CD11b immunefluorescence staining.Firstly,the effects of ISL on microglial viability were explored by the MTT assay.The microglial cells were pretreated by ISL(5μmol·L-1)and then stimulated by LPS(100 ng·mL-1),the production of NO was detected by Griess reagent,the change of microglial morphology was observed by immunefluorescence staining.The production of IL-1βand TNF-αin culture medium were tested by ELISA,and the m RNA expression of pro-inflammatory factors,including IL-1β,TNF-α,i NOS and COX-2 was detected by real time PCR.The protein expression of i NOS and COX-2was detected by Western blotting.RESULTS(1)After purifying,microglial showed the typical morphological characteristics.The cell body was small,protruding outstretched in the branch rod,spindle.After LPS stimulation,the shape was changed to amoeba-like,and the protrusions retracted,cell rounding.In the CD11b immunefluorescence staining,the purity of the microglial was over 98%,and showed powerful phagocytic activity.(2)The results show that ISL(0.16-20μmol·L-1)concentration has no effect on primary microglia′s vitality.And at the concentration of 5μmol·L-1ISL has the significantly inhibitory effect on NO production induced by LPS.(3)The results shows that ISL pretreatment can significantly inhibit LPS-induced microglial activation,and can significantly reduce the production of NO in a dose-dependent manner(P<0.05).When stimulated by LPS for12 h,the expression of TNF-αand IL-1βm RNA in the culture medium are reduced by ISL pre-treatment(P<0.05).After LPS stimulation for 24 h,the expression of COX2 and i NOS m RNA and protein can be reduced(P<0.05).After stimulated by LPS from 0.5 to 2 h,p-ERK expression has be decreased.CONCLUSION We successfully obtain primary microglia with high-purity by culturing in vitro.We find that ISL can significantly reduce the levels of proinflammatory facters,which indicate ISL can inhibit microglia activation and neuroinflammation by blocking ERK1/2 signal pathway.
基金Fourth Chinese Materia Medica Resources Survey(Z135080000022)。
文摘Schisandra Chinensis Fructus(SCF)is the fruit of Schisandra chinensis(Turcz.)Baill.,a perennial vine.It was first recorded in Shen Nong′s herbal classic and has a long application history.Studies have shown that SCF has anti-inflammatory,protective liver,antioxidant,antibacterial and other pharmacological effects.Ancient prescriptions are commonly used in the treatment of chronic diarrhea and other intestinal diseases and diabetes.Modern clinical pharmacology features of SCF polysaccharide(SCFP)in diabetes,liver diseases,enteritis and other aspects have achieved excellent results.Gut is an important digestive organ of human body,but intestinal diseases are varied,including Crohn′s disease,ulcerative colitis,intestinal flora imbalance,etc..It is a chronic and non-specific inflammatory disease.The disease is persisted for a long time and the incidence rate is expected to rise.Most of the symptoms are recurrent diarrhea,bloody stool and abdominal pain.It is considered by the World Health Organization as a refractory disease.At present,there is little possibility of complete cure,which is closely related to complex environmental factors,eating habits and heredity.In recent years,clinical studies have found that SCFP has a variety of pharmacological effects on intestinal protection.①Reduce inflammatory factors:intestinal mucositis is a common adverse reaction in patients with chemotherapy.The development of mucositis is related to pro-inflammatory factors such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),IL^(-1)β,Interferon-γ(IFN-γ).SCFP can significantly reduce IL-6 TNF-α,IL^(-1)β,and IL-8,as well as the accumulation of T cells in the process of resisting apoptosis,reduce the inflammatory reaction and protect the damage to villi and crypts,improve the symptoms of small intestinal mucositis caused by weight loss and diarrhea.②Promote immunoglobulin A secretion:intestinal mucosal immunity is the first line of defense of the body′s immune system.Its main antibody is secretory immunoglobulin A,which can destroy and phagocytize microorganisms,bacteria and viruses.SCFP can improve intestinal immunity by increasing the number and activity of T lymphocytes,promoting the secretion of secretory immunoglobulin A,and affecting the activity of a variety of cytokines.③Regulation of intestinal flora:the flora in the intestine has the functions of auxiliary nutrient absorption,biological antagonism and immune regulation,and can form a natural barrier for the host's intestine.When the human intestinal flora is disordered,probiotics will be greatly reduced,harmful bacteria will proliferate and destroy the intestinal environment.Under these conditions,the intake of SCFP significantly increased the number of beneficial bacteria such as bifidobacteria and lactobacillus,and significantly decreased the number of conditional pathogens such as enterococcus and escherichia coli,indicating that SCFP can indeed regulate the intestinal disorder caused by lincomycin hydrochloride to a certain extent. This may be because beneficial bacteria in the intestine metabolize polysaccharides produce short chain fatty acids such as lactic acid and acetic acid, which reduces the pH value in the intestine and inhibits the growth of enterococcus and Escherichia coli. In conclusion, SCFP can treat and protect intestinal diseases to a certain extent, which provides a favorable basis for the treatment of intestinal diseases.
基金Supported by the National Key R&D Program of China(2017YFD0502200)the Applied Technology Research and Development Plan of Heilongjiang Province(GX18B023)Xinjiang Uygur Autonomous Region Science and Technology Support Project Plan(2020E0236)。
文摘The pathogenesis of animal osteoarthritis(OA)is not clear.The aim of this study was to investigate the dynamic changes of cartilage lesions and serum IL-1β,TNF-α,IL-6 and IL-17 inflammatory factors in a post-traumatic osteoarthritis model in rats,and to explore their values in monitoring the progression of osteoarthritis.Forty male SD rats were randomly divided into the control group(n=20)and the model group(n=20).The OA model was established by anterior cruciate ligament transection combined with partial medial meniscus resection(ACLT+PMMx)in the model group,and sham operation was performed in the control group.On the 0th,10th,20th and 30th days after modeling,the degree of joint swelling and the number of white blood cells in the peripheral blood were evaluated,and tibia samples were taken for macroscopic observation score and pathological observation.The serum levels ofIL-1β,TNF-α,IL-6 and IL-17 concentration were detected by enzyme-linked immunosorbent assay(ELISA).The results showed that the cartilage damage in the tibial surface and pathological observations of rats in the model group gradually increased with time,the eye scores increased and increased significantly on the 20th and 30th days(P<0.01),and the joints of the rats were swollen.And the swelling was extremely significant on the 30th day(P<0.01).Compared with the control group,the number of white blood cells in the peripheral blood of the model group gradually increased,and the increase was extremely significant on the 20th and 30th days(P<0.01).The serum IL-17 concentration in the model group tended to increase,and the increase was significant on the 20th day(P<0.05)and extremely significant on the 30th day(P<0.01);the serum TNF-αand IL-1βconcentrations in the model group showed an increasing trend,and the increase was extremely significant on the 10th,20th and 30th days(P<0.01);IL-6 concentration in the serum of the model group showed an increasing trend,and it increased significantly on the 30th day(P<0.05).The results showed that in the rat model of traumatic OA,the degree of joint damage gradually increased,and the expression of blood inflammatory indexes IL-1β,TNF-α,IL-17 and IL-6 increased.The research provided a basis for prompting and monitoring the timing of OA treatment and the development of the disease in animal clinical.
