My X axis is made up of two motors slaved in Mach4. Motor 0 and Motor 3. I set both of these motors to reverse and restarted MAch4 and nothing happened they moved in the same direction as before.
Then my guess is that you are not using a PMDX-SmartBOB as your motion device (EDIT: or you are but something is wrong)
[EDIT If you are not using a PMDX device...] Look in your motion device's plugin configuration and see if there are settings for motor direction there. Go to the Mach4 "Configure" menu and select "Plugins..." and then click on the name of your motion device (or on older versions of Mach4, click on the "Configure" button next to your motion device).
If you tell us which motion device you have (for example, Darwin parallel port, Ethernet SmoothStepper, PoKeys, etc.), someone here might chime in with specific suggestions. And while you're at it, which version of Mach4 and which plug-in version you are running.
Bob