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AIM:To investigate the expression of gonadotropin-releasinghormone(GnRH)receptor and the effects of GnRH analog(alarelin)on proliferation of cultured gastric smooth musclecells(GSMC)of rats.METHODS:Immunohistochemical ABC methods and in situhybridization methods were used to dectect protein andmRNA expression of GnRH receptor in GSMC,respectively.Techniques of cell culture,OD value of MTT test,measureof ~3H-TdR incorporation,average fluorescent values ofproliferating cell nuclear antigen(PCNA)and flow oltometricDNA analysis were used in the experiment.RESULTS:The cultured GSMC of rats showed immunoreactivityfor GnRH receptor;positive staining was located in cytoplasm.GnRH receptor mRNA hybridized signals were also detectedin cytoplasm.When alarelin(10~(-9),10~(-7),10~(-5) mol/L)wasadministered into the medium and incubated for 24h,ODvalue of MTT,~3H-TdR incorporation and average fluorescentvalues of PCNA all decreased significantly as compared withthe control group(P<0.05).The maximum inhibitory effect oncell proliferation was achieved a concentration of 10~(-5) mol/Land it acted in a dose-dependent manner.Flow cytometricDNA analysis revealed that alarelin could significantlyenhance ratio of G_1 phase and decrease ratio of S phase ofGSMC of rats(P<0.05).The maximum inhibitory effect onratio of S phase was at the concentration of 10~(-5) mol/L andalso acted in a dose-dependent manner.CONCLUSION:Our data suggest that GnRH receptor canbe expressed by GSMC of rats.GnRH analogue can directlyinhibit proliferation and DNA synthesis of rat GSMC throughGnRH receptors.
AIM: To investigate the expression of gonadotropin-releasinghormone (GnRH) receptor and the effects of GnRH analog (alarelin) on proliferation of cultured gastric smooth muscle cells (GSMC) of rats. METHODS: Immunohistochemical ABC methods and in situ hybridization methods were used to dectect protein and mRNA expression of GnRH receptor in GSMC, respectively. Techniques of cell culture, OD value of MTT test, measure of ~ 3H-TdR incorporation, average fluorescent values of proliferating cell nuclear antigen (PCNA) and flow oltometric DNA analysis were used in the experiment .RESULTS: The cultured GSMC of rats showed immunoreactivity for GnRH receptor; positive staining was located in cytoplasm. GnRH receptor mRNA hybridized signals were also detected in cytoplasm. Whilst alarelin (10-9, 10-7, 10-5) mol / L) wasadministered into the medium and incubated for 24h, ODvalue of MTT, ~ 3H-TdR incorporation and average fluorescentvalues of PCNA all decreased significantly compared to the control group (P <0.05). inhibitory effect once11 proliferation was achieved a concentration of 10 ~ (-5) mol / Land it acted in a dose-dependent manner. Flow cytometric DNA analysis revealed that alalin could significantly enhance ratio of G_1 phase and decrease ratio of S phase of GSMC of rats (P <0.05). The maximum inhibitory effect on phase of S phase was at the concentration of 10 ~ (-5) mol / L andalso acted in a dose-dependent manner. CONCLUSION: Our data suggest that GnRH receptor canbe expressed by GSMC of rats. GnRH analogue can directly inhibit proliferation and DNA synthesis of rat GSMC through GnRH receptors.